Effective Date: 
Mon, 04/27/2020

ITS is implementing a program called QuickSupport which allows ITS staff to remotely access a computer to provide requested support.  When an IT ticket is opened, the tech who responds may request that they connect to your computer using QuickSupport in order to diagnose and/or resolve your issue.  Generally, SHS IT issues are reported to internal support staff (Robert Antonino or Cathy Sanders) who handle all issues with the EMR and minor to moderate system software/hardware issues. 


In the event that a SHS staff member needs to contact ITS directly for workstation support, the following procedure must be followed when using QuickSupport:


Procedures for HIPAA Units/Clients

  • Some mechanics of receiving remote IT support:
    • ITS uses a program called QuickSupport for remote assistance.
    • ITS techs will not connect to your computer without your permission. You give your permission by providing your QuickSupport ID and password to the tech. The tech will help you do this. There will also be a screen for you to “accept” the session.
    • Your QuickSupport ID is specific to your computer. A new password is generated each time you launch the program. 
    • Once you connect, the name and phone number of the ITS technician is displayed in a separate small window during the remote control session. Make sure this matches the person on the phone with you. If you don’t have confidence that you are talking to an ITS employee, decline the session, hang up, and contact the Support Center (9-HELP) to verify.
    • Have the tech explain why they are calling before accepting a remote assistance session. The reason should match what the client expects.
  • Tell the tech that they will be working on a HIPAA machine.
  • Inform the tech if the nature of the problem means HIPAA-related data might displayed on the screen. This includes pop-ups from HIPAA-related apps. It's not OK to open or run any HIPAA-related programs during a remote assistance session. If this will be required, remote assistance isn't an option and an in-person visit will need to be scheduled.
  • Close windows and log out of all HIPAA-related sessions (PnC & other health systems, PPS, email for Benefits Office) before accepting a remote support session. No ePHI or HIPAA systems can be open or visible on the computer.
  • Be aware that if your computer is rebooted during a QuickSupport session, the tech might automatically be reconnected.
  • Files containing restricted data of any sort may not be transferred using QuickSupport.
  • Be aware that techs are supposed to confirm required HIPAA security settings when working on a HIPAA machine if this hasn’t been done in the last 90 days. This is in addition to assisting with the specific problem at hand.
  • Always quit QuickSupport when the remote assistance session is complete. The tech should tell you when to do this, but be sure to ask, just to be safe.
Key Points: 
  • ITS can  now support SHS HIPAA systems using a tool called QuickSupport.
  • Specific rules must be followed to ensure protection of SHS HIPAA systems and data.
  • Always quit QuickSupport when the remote assitance session is complete.


Effective Date: 
Wed, 05/16/2018

All students evaluated by Counseling and Psychological Services (CAPS) will be screened for their alcohol and other drug use based on questions during the Initial Assessment and and student responses on the Student Questionnaire.  Psychiatry staff will be screening for alcohol and other drug use during the Psychiatry Intake. 

After the initial visits with Counseling and Psychiatry, students should be re-assessed for alcohol and other drug use annually when the student fills out the Student Questionnaire and when there are clinical signs of alcohol and other drug use.

Psychiatry and Counseling providers should consider urine drug testing for the purposes of safety and diagnosis when there are physical and behavioral signs of alcohol and other drug use.

Although drug testing can be a useful tool for making clinical decisions, it should not be the only tool. When combined with a patient’s history, collateral information from a spouse or other family member (obtained with permission of the patient), questionnaires, biological markers, and a practitioner’s clinical judgment, drug testing provides information that:

Can affect clinical decisions on a patient’s substance use that affects other medical conditions

Can affect clinical decisions about pharmacotherapy, especially with controlled substances.

Increases the safety of prescribing medications by identifying the potential for overdose or serious drug interactions.

Helps clinicians assess patient use of opioids for chronic pain management or compliance with pharmacotherapy for opioid maintenance treatment for opioid use disorders.

Helps the clinician assess the efficacy of the treatment plan and the current level of care for chronic pain management and substance use disorders (SUDs).

Prevents dangerous medication interactions during surgery or other medical procedures.

Aids in screening, assessing, and diagnosing an SUD, although drug testing is not a definitive indication of an SUD.

Identifies women who are pregnant, or who want to become pregnant, and are using drugs or alcohol.

Monitors abstinence in a patient with a known SUD.

Verifies, contradicts, or adds to a patient’s self-report or family member’s report of substance use.
Identifies a relapse to substance use.

-adapted from:  Substance Abuse and Mental Health Services Administration. Clinical Drug Testing in Primary Care. Technical Assistance Publication (TAP) 32. HHS Publication No. (SMA) 12-4668. Rockville, MD: Substance Abuse and Mental Health Services Administration, 2012.




At the initial Counseling and Psychiatry visit, inquire about the student’s alcohol and other drug use.  Review the student’s self-report in the Student Questionnaire.

The establishment of a culturally sensitive and confidential clinical environment is crucial to open discussions on alcohol and other drug use.  Inquire about the student’s beliefs and perception of alcohol and other drug use. 

At subsequent visits, inquire about the student’s alcohol and drug use if the student is exhibiting physical and behavioral signs, e.g., slurring of speech, lack of coordination, responding to hallucinations, or hyperkinetic movements. 

If the student is not willing or capable of discussing alcohol and other drug use, considering ordering a urine drug test to be completed at the Student Health Center laboratory or off-campus.  CAPS staff can ask Psychiatry staff to order a urine drug test.    

After the initial urine drug test, consider ordering urine drug tests during follow-up visits if the student continues to exhibit signs of substance abuse or is not engaging in treatment planning for substance abuse.

Whether students engage or not in treatment planning may involve trust in the provider.  The provider should actively create a confidential clinical environment that encourages honest reporting by the student.  The provider should also ask how the student’s ethnicity and culture may affect his or her understanding of substance abuse and the implications of drug testing. 

Key Points: 

Students should be screened for alcohol and other drug use at their initial visit, yearly, and as clinically indicated. 

Urine drug testing may yield important information to aid diagnosis and to ensure safety.

Urine drug testing should NOT take the place of discussions on how the student is using alcohol and other drugs. 

The establishment of a culturally competent and safe clinical environment by the provider allows for possibility of open discussions on alcohol and other drug use.


Effective Date: 
Mon, 07/24/2017
Mon, 02/05/2018
Mon, 02/05/2018

This policy provides the guideline for the cleaning and disinfecting of the patient care areas in CAPS, as per the SHC's Infection Prevention and Control Program.


The reception/waiting area and the counseling offices are subject to use by students, some of whom may be  ill or carriers of infectious/contagious conditions.  By nature of the patient traffic volume, fomite surfaces in these areas can be contaminated and become sources of infection to other patients and staff.

Basic cleaning is provided by the custodial staff from the Campus Facilities Department.  The cleaning contract specifies that the custodial staff will provide the following weekday services :

  • All restrooms are cleaned using approved cleaning agents/products
  • Floors are swept, mopped or vacuumed
  • Containers of non-hazardous trash are emptied
  • Less frequent cleaning, such as shampooing upholstery or cleaning window blinds is arranged by the SHC manager in conjunction with the Facilities staff

CAPS Reception staff are responsible for cleaning surface areas and equipment used in patient care. This cleaning is done daily, and as needed through out the day. Approved cleaning products are listed on the CAPS Cleaning Log and are in accordance with the SHC's Infection Contol Committee cleaning guidelines.

Daily cleaning includes but is not limited to: horizontal Reception surfaces (such as keyboard, mouse and desk), clipboards and pens, computer touch screens and touch pads, vital signs station equipment.

Private offices and other CAPS areas (such as kitchen, group room, scanning room, etc.) can be cleaned as needed, using the approved cleaning products listed on the log.

Key Points: 
  • In accordance with the SHC's Infection Prevention and Contol Program, daily cleaning in CAPS areas is done using approved cleaning products.
  • See Cleaning Log for details.
Attached File: 


Effective Date: 
Mon, 06/01/2015
Mon, 05/07/2018



 Let's Talk is an outreach program designed to engage students by providing informal walk-in consultations with CAPS counselors at sites across campus.

Let’s Talk is intended to reach students in distress who might be unlikely to seek traditional mental health services at CAPS. This service is called "informal consultation" and is different from formal counseling. One difference relates to its accessibility. There is no clinical paperwork to fill out, no formal intake, no appointments, and no fees. Students are encouraged to drop by and talk about whatever is important to them, much as they might talk with a TA, residence hall director, or academic advisor and students can choose to remain anonymous if they prefer.  Let’s Talk is advertised to students as a 10-15 minute consultation.  However, CAPS staff may use their clinical judgment to lengthen the time of meeting with the student if necessary.  When the meeting lasts much longer than 15 minutes, staff should let the student know that this is an exception, since Let’s Talk is understood to be a brief consultation.

The goal of a Let’s Talk consultation in general is engagement. The clinician listens, empathizes, problem-solves, provides mental health and general health information, conducts informal needs assessments, offers advocacy and referrals, and -- most importantly -- plants the seeds of a relationship to facilitate the student seeking additional help if necessary. It is akin to the kind of pre-counseling conversation one might have with a student after giving a stress management presentation on campus. As such, it is not considered a "clinical" service.

A number of students are served by a one-time consultation. Others benefit from intermittent, as-needed visits. Some are referred to CAPS for treatment following a brief assessment. Having made a positive, informal first contact with a clinician is usually sufficient to mitigate any lingering barriers to accessing mental health services at CAPS.

Though Let's Talk is designed to be a short-term intervention, occasionally a student may visit more than once or twice when barriers are robust. For example, some students need more help than one visit can provide but find accessing services at the health center very uncomfortable. They may need multiple visits at Let's Talk to be ready to accept a referral. However, Let's Talk is not a substitute for regular counseling and should not be treated as such.

Though the primary mission of Let's Talk is to reach students who do not conventionally seek mental health services, many other students come simply because of the convenience and immediacy. A Let's Talk consultation can often head off a crisis before it happens, facilitate a quick referral to CAPS, and, in many cases, prevent the unnecessary use of CAPS intakes for students who need a simple, brief intervention.


Let’s Talk is held at different locations across campus, with emphasis on reaching student communities who may have difficulties accessing traditional mental health services or have a hard time accessing CAPS because of tight schedules or geographic location.  Examples of target of student communities include students of color, first-generation college students, and students in Science, Technology, Engineering, and Math.  Despite the emphasis on hosting Let’s Talk at sites convenient to particular communities, every site is open to all UCSC students. Indeed, many students access a site for other reasons, including fit with one's schedule and interest in talking with a particular counselor. Students find out about counselors by reading their biographies and seeing their pictures on the Let's Talk website.

For a complete list of current Let's Talk sites and times, refer to the CAPS web site: http:


Material Necessary when Conducting Let’s Talk:

Telephone. Most sites provide one for you.
Clock or watch for monitoring time.
Emergency phone numbers: CAPS (9-2628 for the front desk) and UCSC police (9-2345 Emergency; 9-4856 Dispatch).
CAPS laptop with remote access Point and Click
"Let's Talk FAQ"
“Let’s Talk Sign-in Sheet
"Let's Talk Busy Notice"*
"Let's Talk Leaving Early Notice"*
Billboards pointing students to location
5150 procedures, Crisis assessment form, 5150 form, informed consent form to be used only in an emergency.
Site Contact Person for support and to hang notices on door PRN
Advertising Post Card or Brochure

Information to Obtain from Students at Let’s Talk:

Ask students for first and last name at beginning of consultation time, and inform them of option to meet anonymously if they prefer.
After consultation is complete, if counselor has a student’s full name and it seems clinically appropriate, may check the student’s PNC record to see if any other action is warranted.   (For example, it might be appropriate to notify other CAPS staff of the student’s consultation with Let’s Talk. ROI is not needed since let’s talk is part of the CAPS services).


No documentation is required for Let’s Talk consultation meetings with a student.  Remember that let’s talk is considered an informal consultation and not a formal counseling session.
If the Let’s Talk counselor receives information about a student from a third party who has contacted Let’s Talk that may be useful to document in PnC, encourage that person to call the CAPS Crisis Counselor to relay that information.  The CAPS Crisis Counselor can then document in PnC using the Third Party Consultation note.
Let’s Talk counselors do not provide written documentation or case management for students. If a student is requesting written documentation, they should be preferred to phone triage and a formal intake.
Let’s talk counselor collects demographic data about each individual scene for consultation (see demographic sheet) which is aggregated at end of year.

Transferring Care to CAPS:

Once students have engaged with you, they may be interested in becoming a regular client at CAPS.

From the Let’s Talk Meeting, the counselor can schedule the student for a Phone Triage appointment in PnC. In the phone triage block on PnC, please note the student was referred from Let’s Talk.
A student must not be referred directly to an Intake appointment from Let’s talk. They must complete a phone triage because risk questions are not asked during a Let’s Talk contact.
A student may be scheduled for an in-person phone triage (versus phone triage) with the Lets Talk counselor at their office (not at the Let’s Talk location).
If the Let’s Talk Counselor is unable to schedule the Phone Triage appointment (via PNC or by phone with the front office), the student can contact the Central Office to schedule a Phone Triage appointment.

Informed Consent:

Because we do not consider Let's Talk a clinical service, students are not required to sign a "consent for treatment" and "limitations of confidentiality" statement.  Let’s Talk counselors do not have formal conversations about confidentiality at the beginning of the Let’s Talk consultation, as Let's Talk is akin to a conversation one might have with a student after a stress management outreach presentation

Students coming into Let’s Talk are provided a Professional Disclosure statement which describes the service and guidelines for confidentiality, but the student is not asked to sign this form. 
Students are encouraged to read the "Frequently Asked Questions" section of the Let's Talk website, which explains the difference between Let's Talk and formal counseling and outlines the limits of confidentiality. Counselors are also provided with a paper copy of the "Let's Talk FAQ"* to give to students as necessary.
It may be necessary to discuss and clarify the limits of confidentiality with a student utilizing a Let’s Talk consultation in certain circumstances. For example, if a student was on the verge of disclosing something that might require breaking confidentiality, the clinician would stop the process to discuss the issue.

Repeated Visits:

On average, the yearly average of Let’s Talk visits is one-to consultations per year.  Occasionally, a student may be reluctant to accept a referral to more formal counseling.

If a student continues to use Let’s Talk repeatedly (more than 2x per year):

Gently discourage the student from using Let’s Talk as a substitute for counseling
If a period of more extensive engagement with Let’s Talk let’s talk may be warranted, please consult with a member of the Let’s Talk team or management.

Assessing Risk:

Students who utilize Let’s Talk are not asked directly about suicide unless clinical judgment indicates it is warranted. Let’s Talk is not a clinical service and its goal is to provide access to students who may be reluctant to seek traditional mental health services because of its associated stigma.
For students who voluntarily reveal imminent risk, the student is referred to the Central Office for assessment in same day Crisis Services and asked to walk to the Central office at that time (the Let’s Talk counselor should call ahead and alert the on-call CAPS clinician).
In rare cases, the Let’s Talk Counselor may need to walk the student over, or call the Central Office for a CAPS clinician to come to the Let’s Talk location to assist in an on-site assessment.  Calling the campus Police is also an option in urgent cases.


If the student is psychotic, actively suicidal or homicidal, or might warrant emergency help for other reasons, the following options can be utilized:

If the student needs immediate counseling follow-up but does not need hospitalization. Make a short-term safety and coping plan with the student and schedule a Priority CAPS Intake.
If the student may need hospitalization. If you are concerned that the student needs to be hospitalized:

Call the CAPS Central Office for consultation with management
The Let’s Talk counselor may need to walk the student over to the CAPS Central Office of Campus Police can be contacted for transport
Call the Central Office for a CAPS clinician or management to come to the Let’s Talk location to assist in an on-site assessment and possible hospitalization. .

Unexpectedly Leaving Early from Let’s Talk:

If you have to leave the site early, either to facilitate a hospitalization or for any other reason, leave a copy of the "Let's Talk Leaving Early Notice" on the door.

Absences from Let’s Talk:

Planned absences:

Contact your site to inform them you will be absent and forward a copy of the "Let's Talk Absence Notice" *.  Please have your contact person post the notice on the door of your site during the scheduled Let’s Talk hours

Unplanned absences:

When you call the CAPS front desk to inform them you will be out for the day, please ask them to contact your Let's Talk site. The front desk has a list of all sites and contact names.

Front desk guidelines for unplanned absences:

In the event a Let's Talk clinician calls in sick, please call the person listed under "contact in the event of absence" on the "Let's Talk Master List"* and inform them that Let’s Talk” will not be offered that day.

Follow up with an e-mail with the "Let's Talk Absence Notice"* attached. The contact person should post this on the door.

Responsibilities of Campus Offices Which Host Sites:

Provide an office for the scheduled Let’s Talk time;
Provide a phone, unless arrangements have been made for you to use your own cell phone
Have someone nearby in case of an emergency.  It's occasionally helpful to remind site personnel of this need; we can be easily forgotten in our offices as people go home for the day.

Let's Talk Documents:

All documents are also located in the "Let's Talk Administration" folder on the K drive.  Since the K: drive is not accessible at Let’s Talk sites, it is important that you bring these materials with you to the site.


Effective Date: 
Tue, 03/25/2014
Tue, 02/04/2020

Purpose and Scope:
To describe the steps for manually entering and approving lab results in the LIS (Orchard Harvest).





  1. Log on to Harvest Orchard by selecting your username from the drop down box. Enter your password in the appropriate space.
  2. Select the Work in Progress box from the Work Center menu. (Alternatively, Work in Progress can be selected from the drop down menu under Laboratory).
  3. Double click on the patient’s test that is to be resulted. Enter the appropriate numerical result in the Result column or select a text result by clicking the down arrow on the Text Result column.
  4. Click the Approve button on the top right of the screen. This will release results to the medical record.
  5. If results are present from an instrument interface, review results for accuracy, enter any additional results or comments and click the Approve button to release to the medical record.
Key Points: 


Orchard Harvest LIS Help Manual Version 8.5  Copyright 2009


Effective Date: 
Thu, 10/17/2013
Wed, 02/05/2020
Tue, 04/17/2018

The clinical laboratory generates materials that are considered hazardous and/or biohazardous waste. The handling of and storage of hazardous materials and hazardous waste will be monitored regularly according to University requirements.

The clinical laboratory completes and maintains a "Storage Area Self-Inspection Form" for Hazardous Materials/Hazardous Waste. The form is completed weekly. The form includes: Inspection date, Area free of spills or leaks, Proper secondary containment, Contaminant clean, Containers labeled, Incompatibles segregated, Waste tagged and dated, Waste within storage time, Emergency procedure posting, and Inspectors initials and Comments/Corrective Actions.

Staff are trained on proper identification, management, handling, transport, and disposal of all hazardous materials and wastes.



Hazardous Waste

Wright Stain and Gram Stain washings are considered hazardous waste. The washings are collected in e-tagged (UCSC online hazardous waste program WASTe) leakproof containers and stored until two thirds full or 90 days whichever comes first. They are subsequently picked up by UCSC EHS department.

Leftover reagents from the Sysmex XN-430 analyzer are not considered hazardous and may be diluted with water and disposed of down the drain.

Biohazardous Waste

The biohazardous waste the lab generates consists of blood tubes and any visibly bloody tissues, papers or containers. Blood tubes are disposed of in sharps containers and then deposited in the biohazardous waste receptacle located in the lab storage closet. Bloody materials are put in the red bags lining the self closing waste cans under the counters in the lab. The red bags are then deposited in the biohazardous waste receptacle located in the closet.



Effective Date: 
Mon, 09/23/2013
Wed, 02/05/2020
Wed, 10/23/2019


The quality assurance plan outlines written policies, procedures and activities designed to monitor and evaluate the quality of testing processes (preanalytic,analytic and postanalytic). Policies and procedures insure accurate reliable and prompt reporting of test results, as well as help to meet standards from regulating agencies. Ongoing quality assurance activities can detect errors, procedural lapses or divergences from goals, while also suggesting changes need in procedures or training.

The plan includes policies and procedures that describe quality control, proficiency testing, personnel training and competencies, management of test results, handling of test specimens, and the verification of accuracy of instrumentation and methodologies.

The plan also includes activities to detect prompt result reporting, to detect mishandling of specimens and to monitor the proficiency of testing personnel.

Quality Control

Controls are used according to manufacturer’s recommendations or as required by accrediting agencies for each testing procedure performed. The medical director designates the lab supervisor to monitor and review all quality control data and/or charts on a timely basis. See the QUALITY CONTROL policy located in Section VI of the LAB MANUAL for specific descriptions of quality control materials and activities.


Proficiency samples are provided by the American Proficiency Institute (API) on a quarterly schedule. Proficiency samples are handled in the same manner as patient samples. All testing personnel participate in analyzing proficiency samples. The medical director reviews all results and corrective actions (if needed). See the PROFICIENCY TESTING policy located in Section VI of the LAB MANUAL for more details of proficiency testing activities and monitoring.

Training and Competency

All personnel are required to undergo an extensive training period before performing testing. Training checklists and performance reviews are reviewed by the lab supervisor. Performance reviews and competency checks are performed at 6 months for new staff, and annually thereafter. Moderate to highly complex testing is only performed by licensed clinical lab scientists, while waived testing is only performed by personnel that have successfully completed training. Phlebotomy is only performed by licensed phlebotomists or clinical lab scientists. See the LABORATORY STAFF ORIENTATION, TRAINING AND ASSESSMENT policy located in Section VI of the LAB MANUAL for more details.

Test Results

All test results performed in-house are reviewed and released by clinical laboratory scientists. Abnormal or unexpected results are verified as needed. Any results questioned by the clinician are repeated and verified if possible. Critical results are called to clinicians within 30 minutes of final result. Critical result phone calls follow the format of using two forms of patient identification and having the recipient read-back the result. Documentation of the call is included in the medical record with the result. Corrected test results are called to the clinician as soon as possible. Documentation of corrected result calls is included in the medical record with the results. See the REVIEW OF TEST RESULTS policy located in Section VI of the LAB MANUAL for more details.

Lab Specimens

All lab specimens are labeled with Point and Click (MIS) labels. Hand labeled specimens may be acceptable but must have at least two forms of identification.

Unlabeled or inadequately labeled specimens are rejected and discarded by Lab staff.  In this case, the Lab Supervisor notifies the Medical Director, or their designee, who messages all clinicians potentially affected by the discarded specimen.

Compromised specimens (clotted or hemolyzed) may be recollected if possible.

All specimens with lab orders are accessioned into Point and Click (MIS) which sends them to Orchard Harvest (LIS) system.


Accuracy of Instruments and Methods

All instruments and methodologies are validated before being adopted into use for routine patient testing. The validation consists of precision testing, correlation with a previously verified method and verification of linearity (if quantitative results are involved). Established instruments and methods are verified to be accurate through the use of Interlaboratory Quality Assurance Programs (IQAP) when available, quarterly proficiency testing and biannual calibration verification (where applicable). See procedures, EVALUATION OF AUTOMATED TEST METHODS, LINEARITY TESTING (REPORTABLE RANGE) CALIBRATION VERIFICATION, PROFICIENCY TESTING and QUALITY CONTROL AND ASSESSMENT in section VI of the Laboratory Manual for further information.



Quality assurance activities are the ongoing studies or monitoring of lab processes in order to measure the performance or effectiveness of protocols in relation to goals set. These activities may be ongoing or may change periodically to meet the needs and the goals of lab management.

Turn around times

Turn around times or TAT is usually perceived as the time it takes once a specimen is received in the lab to the time a final result is generated. It is desirable to decrease the TAT for testing that is needed urgently (STAT) so as to expedite patient care. Monitoring TAT can help facilitate corrective actions where the TAT does not meet goals and can lead to overall changes in procedures and personnel in order to maximize efficiencies. The lab will monitor STAT in-house CBCs and STAT urine dipsticks for TAT during the current year.

Unlabeled or mislabeled specimens

Unlabeled or incorrectly labeled specimens will be discarded by Lab staff. The Lab Supervisor will notify the Medical Director, or their designee, who will message all clinicians potentially afffected by the discarded specimen.

Accuracy of Test Results

Inaccurate or erroneous lab results can lead to serious errors in patient care. Erroneous lab results can be caused by typographical errors, mislabeled specimens and/or instrument malfunction. All erroneous lab results must be corrected and the clinician notified promptly. Such errors are documented in the problem log.

Quality Improvement

Projects are to be undertaken to assess the quality of certain processes and procedures during the course of the year. The projects will ascertain to measure the quality of the process in terms of sensitivity, specificity or timeliness. It will also be assessed as to whether the process can be improved by implementing various changes or having staff focus on the issue. Examples of such past QI projects are the Wet Mount vs Trichomonas Culture project (improve sensitivity) or the  Urine Leukocyte Esterase vs Manual WBC Count project (improve accuracy, sensitivity and utilization).


Effective Date: 
Fri, 02/01/2019
Tue, 02/04/2020
Tue, 01/29/2019



The Sysmex XN-450/XN-430 is a quantitative automated hematology analyzer for in vitro diagnostic use in determining 23 whole blood diagnostic parameters (WBC, RBC, HGB, HCT, MCV, MCH, MCHC, PLT, RDW-SD, RDW-CV, MPV, NEUT#, LYMPH#, MONO#, EO#, BASO#, IG#, NEUT%, LYMPH%, MONO%, EO%,BASO%, IG%).  Examination of the numerical and/or morphological findings of the complete blood count by the physician are useful in the diagnosis of disease states such as anemias, leukemias, allergic reactions, viral, bacterial, and parasitic infections.

This device performs hematology analyses based on the hydrodynamically focused impedance measurement, the flow cytometry method (using a semiconductor laser) and the SLS-hemoglobin method.

The device counts and sizes red blood cells (RBC) and platelets (PLT) using hydrodynamic impedance counting.  At the same time the hematocrit (HCT) is calculated via the RBC pulse height detection method.   Cytometry is used to analyze physiological and chemical characteristics of cells and other biological particles. Flow cytometry is a method used to analyze those cells and particles as they pass through extremely small flow cells.

Specimen Collection and Handling

WARNING:  All patient specimens should be considered potentially infectious and must be handled with precautions used for human blood, as described in CDC recommendations and in compliance with the Federal OSHA Bloodborne Pathogen Standard, 29CFR part 1910.1030.  Follow specimen handling as outlined by laboratory safety policy.

Recommended:   Wear gloves and a lab coat.  Wear safety glasses if there is a risk of splashing.

A. Required specimen

  1.  Whole blood should be collected in EDTA-2K or EDTA-3K anticoagulant.

B.  Specimen volumes required

  1. Optimal draw is a 13 x 75 tube filled to capacity
  2. A minimum of 1 mL whole blood is required for sample analysis
  3. Manual analysis whole blood mode
  • Closed tube – 1 mL
  • Open tube – 300 μL

C.  Unacceptable specimens including those listed below must be redrawn:

  • Clotted samples or those containing clots, fibrin strands, or platelet clumps.  All specimens will be checked visually for obvious clots prior to sampling by the analyzer.
  • Grossly hemolyzed samples.
  • Samples drawn above an IV line.

D.  Characteristics that may affect test results: lipemia, icterus, and cold agglutinins.

E.  Stored Specimen Stability

  •  EDTA blood samples should be analyzed within 4 hours.
  • If analysis cannot be performed as instructed in the Sysmex Instructions for Use, store in refrigerator at 2-8°C.
  • Allow refrigerated samples to come to room temperature and mix well before analysis.
  • Do not place CBC and Diff samples on a mechanical rocker.  Constant rocking may alter white cell membranes, resulting in false interpretive messages.



Equipment Performance Parameters
The SYSMEX 430-XN Analyzer has operating parameters of ambient temperature 15-35 deg C,  of humidity 20-85%.


A.  Supplies

  • Lint-free lab wipes
  • Gauze
  • Test tubes
  • Pipettes
  • Sysmex reagents
  • Commercial controls: XN-L CHECK TM

B.  Sysmex Reagents

  • Sysmex reagents and CELLCLEAN AUTO are used on all Sysmex XN-L Series modules.
  • All reagents are used at room temperature and are to be used within the manufacturer’s expiration date on each container.
  • Record date received and date opened on container.
  • All reagents are azide free and are intended for in vitro diagnostic use only.  Do not ingest.

              XN-L REAGENTS                                    OPEN EXPIRATION

               CELLPACK DCL                                            60 Days

               SULFOLYSER                                               60 Days (1.5L)

               LYSERCELL WDF                                           90 Days

               FLUOROCELL WDF                                         90 Days


C.  Diluents

1.  CELLPACK DCL: Whole blood diluent for use in hematology analyzers.                         CELL PACK STORAGE

  • Store at 2o-35oC away from direct sunlight.
  • If frozen, thaw and mix thoroughly before using.
  • CELLPACK DCL is clear and colorless.  If it is showing signs of contamination or instability such as cloudiness or discoloration, replace container.

                  CELLPACK DCL Stability

  • Unopened, it is stable until expiration date printed on the container.
  • Opened, stable for 60 Days.

                  CELLPACK DCL Hazard Risk

The OSHA Hazard Communication Standard of 29CFR part 1910.1200 requires SDS documentation of ingredients which have been determined to be health hazards, comprise 1% or greater of the composition, are physical hazards, are capable of release to exceed permissible exposure limit/threshold limit values or have been identified as carcinogens.  CELLPACK does not have ingredients with those characteristics.




D.  Lysing Reagents

  1. SULFOLYSER (SLS): Reagent for the automated determination of hemoglobin concentration of blood.  Sulfolyser is lysing reagent that releases the hemoglobin to be measured by the SLS hemoglobin method.

      SULFOLYSER Storage

  • Store at 1o-30oC away from direct sunlight.
  • Allow the container to equilibrate to environmental temperature (15-35o) prior to use.
  • Replace the reagent if it is showing signs of contamination or instability such as cloudiness or discoloration.

         SULFOLYSER Stability

  • Unopened, it is stable until expiration date printed on the container.
  • Opened, stable for 60 Days (1.5L) or 90 Days (5L).

         SULFOLYSER Hazard Risk

The OSHA Hazard Communication Standard of 29CFR part 1910.1200 requires SDS documentation of ingredients which have been determined to be health hazards, comprise 1% or greater of the composition, are physical hazards, are capable of release to exceed permissible exposure limit/threshold limit values or have been identified as carcinogens.  SULFOLYSER does not have ingredients with those characteristics.


2. Lysercell WDF: Reagent product to be combined and used with Fluorocell WDF.  By hemolyzing red blood cells with Lysercell WDF and dying the white blood cell component with Fluorocell WDF, the counts and percentages of neutrophils, lymphocytes, monocytes, eosinophils and basophils are analyzed.

         Lysercell WDF Storage

  • Store at 2o-35oC away from direct sunlight.
  • Use at an environmental temperature (15-35o).
  • Do not use the reagent if it is suspected to have frozen.
  • Replace the reagent if it is showing signs of contamination or instability such as cloudiness or discoloration.

        Lysercell WDF Stability

  • Unopened, it is stable until expiration date printed on the container.
  • Opened, stable for 90 Days.

        Lysercell WDF Hazard Risk

The OSHA Hazard Communication Standard of 29CFR part 1910.1200 requires SDS documentation of ingredients which have been determined to be health hazards, comprise 1% or greater of the composition, are physical hazards, are capable of release to exceed permissible exposure limit/threshold limit values or have been identified as carcinogens.  Lysercell WDF does not have ingredients with those characteristics.

E.  Staining Reagents

1.  Fluorocell WDF: Used to stain the leukocytes in diluted and lysed blood samples for determination of differential count in blood.

        Fluorocell WDF Storage

  • Store at 2o-35oC in a dark place.
  • Do not use the reagent if it is suspected to have frozen.
  • Fluorocell WDF Stability
  • Unopened, it is stable until expiration date printed on the container.
  • Opened, stable for 90 Days.

       Fluorocell WDF Hazard Risk

The OSHA Hazard Communication Standard of 29CFR part 1910.1200 requires SDS documentation of ingredients which have been determined to be health hazards, comprise 1% or greater of the composition, are physical hazards, are capable of release to exceed permissible exposure limit/threshold limit values or have been identified as carcinogens.  Refer to the SDS.

F.  Cleaning Agent

CELLCLEAN AUTO: Detergent for fully automated hematology analyzer.  To be used as a strong alkaline detergent to remove lysing reagents, cellular residuals, and blood proteins remaining in the hydraulics of the analyzer on XN series/XN-L series automated hematology analyzers.

         CELLCLEAN AUTO Storage

  • Store at 1-30o C, away from direct sunlight.
  • Do not use the reagent if it is suspected to have frozen.

         CELLCLEAN AUTO Stability

  • Unopened, it is stable until expiration date printed on the container.

         CELLCLEAN AUTO Hazard Risk

     The OSHA Hazard Communication Standard of 29CFR part 1910.1200 requires SDS documentation of ingredients which have been determined to be health hazards, comprise 1% or greater of the composition, are physical hazards, are capable of release to exceed permissible exposure limit/threshold limit values or have been identified as carcinogens.  Refer to the SDS, CELLCLEAN AUTO is corrosive and may cause burns to skin.

G.  Commercial Control Material for XN-430 Analyzers

  1. XN- L CHECK
  • Manufactured by Streck, available as a tri-level package.
  • Whole blood commercial control used to monitor performance of all XN-450/XN-430 analyzer platforms.
  • Formulation
  1.  XN-L CHECK consists of human and/or animal red and white blood cells with a platelet component suspended in fluid medium.
  2. Each vial contains 3 mL of control material.
  • Storage
  1. Store vials at 2-8oC.
  2. Do not freeze or expose to excessive heat.
  • Stability
  1. Unopened and properly stored, XN-L CHECK is stable until the expiration date printed on the unopened vial.
  2. Open vial stability is 15 days for XN-L CHECK  when promptly refrigerated after each use.
  3. Record the date on each vial upon opening or cap piercing.
  4. Heat or freezing can damage XN-L CHECK without gross visible changes.  Moderate hemolysis can be normal.  Deterioration is suspected when the mean of the control results is not within the assay expected ranges after appropriate troubleshooting.
  5. If deterioration is suspected, call the Sysmex Technical Assistance Center.  1-888-879-7639 (1-888-8SYSMEX)


The human blood used in XN-L CHECK is non-reactive for Hepatitis B Surface Antigen and negative for antibodies to HIV-1, HIV-2, and Hepatitis C Virus using FDA specified techniques.  However, no current tests can assure the absence of these pathogens.  XN-L CHECK should be considered potentially infectious and must be handled with precautions used for human blood as described in CDC recommendations and in compliance with the Federal OSHA Bloodborne Pathogen Standard, 29CFR, 1910.1030.

H.  XN-430 Reagent Replacement

  1. When the replacement of reagent is required, an error message appears. Promptly acknowledge the error message by clicking execute to enter the reagent replace dialog box and proceed to replace the indicated reagent. Verify that “CAPS LOCK is off.
  2. Replacing a new diluent / hemolytic agent
  • Touch the name of the reagent to be replaced.
  • Place a check-mark next to ‘Replace the reagent,’ then place the cursor in the reagent code text box. 
  •   Using the hand-held reader, scan the reagent code on the new reagent container. NOTE: Scan Reagent Code 2 which is on the top of the container.
  • Remove the cap from the expired/empty continer and carefully remove the spout.
  • Pull out the dispensing set straight up.
  • Insert the dispensing set straight into the new reagent container and close the cap.
  • Select [Execute]
  • Reagent replacement starts.  When complete, the dialog bos closes automatically.

3.  Replacing Dye

  • Display the [Reagent Replacement] dialog box.
  • Prepare the new reagent cartridge.
  • Confirm the reagent has not expired.
  • Pull out the dye holder drawer.
  • Slowly remove the dye cover, taking care that dye does not drip.
  • Remove the entire dye holder.
  • When the dye holder is removed, a Help dialog box appears in the IPU screen.
  • Remove the old reagent cartridge from its holder.
  • Install the new reagent cartridge into the holder.
  •      Make sure the color of the label on the new reagent cartridge matches the color  of the dye cover and install.  Analyzer will beep as confirmation of new reagent installation.
  •       If the wrong reagent is installed, the analyzer beeps repeatedly and the Help dialog box appears in the IPU screen.
  • Place the dye cover.
  •        Place into dye holder drawer.
  •        The ID of the new reagent is read automatically and the information is registered.
  • Close the dye holder drawer.
  •        Reagent replacement starts.
  •        When complete, the reagent replacement window closes automatically.


The process for checking the accuracy and precision of an analyzer at regular timed intervals is known as calibration verification.  Historically, a calibrator has been used on a semi-annual basis as a "check" of daily QC procedures.  Since our instrument uses a BeyondCare Quality Monitor application, calibration is verified each time QC is analyzed and the results fall within algorithm specification guidelins developed for the BeyondCare Quality Monitor program.  This means that calibration verification will be verified every 24 hours.  In the "Intended Use" section of the control package inserts, it states that the controls are intended for quality control and calibration verification.  If physical recalibration is necessay, Sysmex technicians will come to the lab and perform the calibration on site.

BCQM supplies the laboratory with a laptop which can be accessed to provide calibration history at any time.  QC and Calibration Verification will be checked by the Supervisor weekly


QC Frequency
Test 3 levels of QC samples once per day of testing using XN-L Commercial Control, manufactured by Streck.

QC Procedure

  • Remove vials from refrigerator and allow them to come to room temperature (18-25oC), for approximately 15 minutes.
  • Mix vials by rolling each vial between the palms of the hands for 15 seconds.
  • Invert each vial by holding the vial at the ends between the thumb and finger.
  • Invert end-over-end using a very quick turning motion of the wrist approximately once a second for one minute.
  • Perform a close visual inspection of each vial confirming the cell button is completely removed from the bottom of the vial and cellular elements are uniformly suspended with no aggregates.
  • Continue inversions as needed to re-suspend the vial contents.
  • Using the mouse, choose manual mode.
  • Barcode the control.
  • Press OK.
  • Place in tube holder and press the transport button.
  1. Check the analysis results in dialog box.
  2. If analyzer displays GREEN, this indicates QC passed and analyzer is ready to process samples. Touch [Accept].
  3. If analyzer displays YELLOW, more information is needed or QC is overdue. The resolve button becomes active if there is a QC value outside of limits.
  4. Resolve is activated: If a QC error has been detected, resolve button becomes active and dynamic troubleshooting prompt guides the end user to the next course of action. The instructions button gives details on how to perform the troubleshooting action.
  5. QC is overdue: End user needs to analyze QC since it exceeds the timeframe from the preferences screen.
  6. If analyzer displays RED, QC failed analysis and analyzer is determined out of service with a reference to a service call.

For a calendar view of whether the QC passed or failed, access the Summary report in the BCQM lap top (from the dashboard icon) which will also display the background status.  Along the bottom of the screen, the codes appearing in the chart boxes will be explained.

New QC Lot crossover or parallel studies

Five days before the last tubes of the current lot expire, the new lot is analyzed two times/day in conjunction with the current QC. The BeyondCare Quality Monitor program establishes the target and limit values for the new QC lot as soon as the first vial of each level gets analyzed.

Reviewing Quality Control Results in BeyondCare Quality Monitor

  • If analyzer displays GREEN, this indicates QC passed and analyzer is ready to process samples.
  • If analyzer displays YELLOW, more information is needed or QC is overdue.
  • More information is needed: If a QC error has been detected, a dynamic  troubleshooting prompt guides the end user to the next course of action. A video display is also a selectable feature.
  • QC is overdue: End user needs to analyze QC since it exceeds the timeframe from the preferences screen.
  • If analyzer displays RED, QC failed analysis and analyzer is determined out of service with a reference to a service call.

Recording and Storage of QC Data

The BeyondCare Quality Monitor application stores the last 2.5 years of QC data on demand. All QC data older than 2.5 years is archived.

Review of QC Data

The following reports will be reviewed biweekly by the Supervisor: Insight Report, Detailed Daily Verification Report, and Continuous Calibration Verification Certificate.

Once these reports have been reviewed through the BeyondCare Quality Monitor application, they can be accessed by going to Activity - Reviewed Documents.
Reports are archived for 2.5 years in BeyondCare Quality Monitor. Reports that are older than 2.5 years can be attained by contacting Sysmex Technical Assistance Center.




Start-Up Procedure

1.  Checks prior to turning on:

  • Visual inspections of analyzer / system / reagents
  •        Verify network/host connections are working properly
  •        Verify sufficient reagent supply is nearby.

2. Turning ON the entire system

  • Verify that all power switches for the device are in the ON position
  • Press the Green power buttton on the front on the analyzer to power ON the entire system

3.  Log on to the XN-430 IPU

  • When the logon dialog box appears, enter the user name as 'm145m'.

4.  Analyzer self checks

  • XN-430 :  Initialization of the mechanical parts: Rinse; Temperature stabilizaion; Background Check (up to 3 times)  Background check results are found in  the BCQM laptop.

5.  Analyze Control Material

Patient Sample Processing

  1. Manual Analysis
  • Check the status of the analyzer.  Confirm the analyzer is ready.
  • If the sample tube holder is not ejected, press the sampl tube holder open/close switch
  • Select Manual Analysis button the the control menu.
  • Scan the barcode on the tube or sample ID can be entered maually.
  • Select OK.
  • Properly mix the specimen and place in tube holder
  • Press the switch on the analyzer
  • The tube holder will slide in and the sample will be aspirated
  • When the analysis is complete, the tube holder slides out.
  • Remove the sample, repeat steps for additional samples
  • Review the IPU to determine whether repeat or reflex testing is required.
  • Rerun sample if required.  Make smear if required.



Test Results

WBC - White Blood Cell or leukocyte count
              LY#          Lymphocyte number
              LY%         Lymphocyte percent (or ratio)
              MO#         Mononuclear cell number
              MO%        Mononuclear cell percent (or ratio)
              EOS#        Eosinophil number  
              EOS %       Eosinophil percent (or ratio)
               BASO#      Basophil number
               BASO %     Basophil percent (or ratio)
               IG#            Immature granulocytes number
               IG%           Immature granulocyte percent (or ratio)
               GR#          Granulocyte number
               GR%         Granulocyte percent (or ratio)
RBC - Red Blood Cell or erythrocyte count
Hgb - Hemoglobin concentration
Hct - Hematocrit (relative volume of erythrocytes)
MCV - Mean Corpuscular (erythrocyte) Volume
MCH - Mean Corpuscular (erythrocyte) Hemoglobin
MCHC - Mean Corpuscular (erythrocyte) Hemoglobin Concentration
Plt - Platelet or thrombocyte count
RDW - Red Cell (erythrocyte volume) Distribution Width
MPV - Mean Platelet (thrombocyte) Volume

Flagged Results



Exceeds linearity

Dilute and re-run


<4.0 or  >15,000

Slide review


<100,000  or >500,000

Slide review


<75 or >105 fl

Slide review


> 36

Check for cold agglutinin by placing in 37C incubator for intervals of 15 min; check for lipemia, hemolysis.



Slide review

Neut #

<1.0 or >8.0

Slide review

 Neut %


Slide rev. for bands

Bands, nRBCs

>5%  on slide rev.

Manual differential

Metas, Myelo, Promylocytes, Bands   Manual differential
Eosinophils >10% on slide rev. Manual differential
Basophils >5% on slide rev. Manual differential


> 50.0

Slide review



Slide review

IG flag   Slide review
Left Shift?   Slide review
Blasts/Abn Lympho?   Slide review - Manual diff if Blasts are seen
Atypical Lympho?   Slide review
NRBC?   Slide review -  If more than 5 NRBC's/100 WBC's - Correct the WBC count See Manual Diff Review


Results over range for Plt, WBC, RBC, HGB, HCT GRAN, LYM

 Make a dilution with normal saline


<50 or >130

Use slide rev to verify - order RBC Morphology and record significant morphology seen.

Reportable Ranges
The Linearity Limit listed below is the range of results over which the Sysmex XN-430 instrument displays, prints and transmits results.  Parameters that exceed these limits are flaggged with @ beside the result.  The sample must be diluted, rerun and multiplied by the dilution factor. Note the use of dilution for linearity on the patient report.


Linearity Limit/

Reportable Range




x 103 cells/ mL



x 106 cells/m L






x 103 cells/ mL

Critical Values

Critical/Alert Values are those results demonstrating such variance from normal as torepresent a pathophysiological state with potential of being life threatening unless action is taken quickly. These results must be immediately reported to the care provider and be documented in the test record as to who was contacted, the time of contact, the person making contact, and that the results were read back.

WBCs K/mm3 <3.0 or >15.0
HGB g/dl <8.0 or > 20.0
HCT % <25 or >60
Platelets  K/mm3 <100 or >600

Laboratory Reference Ranges



Our Reference Ranges


x103 cells/ uL

<21 yrs 4.5-13.2

>21 yrs 3.4-10.0


x106 cells/ uL

Males 4.4-5.9

Females 4.0-5.2



Males 13.5-17.5

Females 12.0-15.5



Males 41-53.0

Females 36-46.0











x103 cells/ uL




<21 yrs 1.0-6.1

>21 yrs 1.0-3.4



<21 yrs 0-1.4

>21 yrs 0-0.8



<21 yrs 1.8-8.0

>21 yrs 1.8-6.8

EOS x109/L 0-0.24
EOS % 0.2-3.0
BASO x109/L 0-0.2
BASO % 0-1.0

Limitations of the Procedure

 K2EDTA is the recommended anticoagulant. K3EDTA is also acceptable. Use of other anticoagulants can yield misleading results.


Interfering Substances

  • Specimens must be free of clots and fibrin strands.
  • Marked changes in plasma constituents (e.g., low sodium, extremely elevated glucose) may cause cells to swell or shrink.  The blood to anticoagulant ratio is important.
  • Red cell fragments, microcytic RBCs or white cell cytoplasmic fragments may interfere with automated platelet counts.
  • Cold agglutinins produce spurious macrocytosis, elevated MCHs, MCHCs, falsely decreased RBC counts and HCTs.  Rare warm agglutinins produce the same spurious results as a cold agglutinin.
  • Extremely elevated WBCs may cause turbidity and falsely increase the hemoglobin, in addition to RBC and HCT values.
  • Severely hemolyzed samples (in vitro) falsely decrease RBC and hematocrit.  Recollect hemolyzed specimens.
  • Giant platelets and clumped platelets may falsely elevate the WBC count and falsely decrease the platelet count.  Platelet clumping and/or "platelet satellitism" can occur in specimens collected in EDTA.  This may falsely elevate the WBC count and falsely decrease the platelet count.   Vortex and reanalyze.
  • Severely icteric samples may falsely elevate the HGB value and related indices.  Make a 1:5 dilution with CELLPACK DCL.
  • Rocking specimen excessively, may affect the WBC differential.
  • Megakaryocytes may falsely increase WBC counts on automated hematology analyzers.


XN-430 Shutdown -performed daily

  • Confirm analyzer and sample unit are at ready
  • If sample tube holder is not ejected, press the sample tube holder open/close switch
  • If any tubes remain in holder, remove.
  • Touch [Menu] on Toolbar.
  • Touch [Shutdown]. Touch [OK]
  •        XN on-board maintenance history will auto-populate Shutdown
  •        IPU will automatically shut off at the conclusion.
  •        Press Green power putton to restart IPU

XN-430 Routine Cleaning - perfomed weekly

CellClean Auto is used to shut down the entire system.  Refer to the XN-L Series Troubleshooting Manual for detailed, illustrated procedures.

  • Confirm analyzer is at ready.
  • Touch the [Maintenance] icon in the Menu screen.
  • Touch [Rinse Instrument].
  • Touch [Routine Cleaning]
  •        If sample tube holder is not ejected, press the sample tube holder open/close  switch and place CELLCLEAN AUTO in tube holder.
  •         Press start switch.
  •         XN-430 on board maintenance will auto-populate Routine Cleaning.


  • Use 1 vial of CELLCLEAN AUTO for a single cleaning.  Do not reuse CELLCLEAN AITO that has previously been used
  • During Shurdown, other sample tubes are not accepted.


Key Points: 


Use 'XN-530/XN430/XN-330 Basic Operation' or 'Beyond a Better Box Quick Guide' for:

  • How to safely use the analyzer
  •  Getting started and running the instrument day‑to‑day
  • Reviewing unusual results (how to read a result report and what flags mean)
  • Performing  procedures such as cleaning, or replacing a reagent

Use the XN-530/XN-430/XN Troubleshooting for troubleshooting problems with your instrument.

Use 'XN-530/XN-430/XN330 General Information' for:

  • What the instrument does and methods it uses
  • Instrument specifications and requirements

Use 'BeyondCare Quality Monitor User Manual' and the 'Beyondcare Quality Monitor Quick Guide' for understanding the quality assurance program:

  • Access and Login
  • Setup
  • Dashboard
  • Activity Tab
  • Reviewed documents
  • Manage QC
  • Calibration History


  1. Sysmex XN-450/XN-430 Automated Hematology Analyzer CLSI Procedure.  Document Number 1235-LSS, Rev. 4, September 2018
  2. Beyondcare Quality Monitor User Manual. Document number: 1346MKT, Rev. 1, June2017



Effective Date: 
Mon, 08/12/2013
Tue, 02/04/2020


Purpose and Scope:

The bacitracin susceptibility test is used to distinguish Group A streptococci, from other streptococci. When grown on blood agar, Group A streptococci are sensitive to (killed by) the antibiotic bacitracin. A sterile disk impregnated with bacitracin (also known as “Taxo A disk”) is placed on the first sector of an isolation plate before incubation. A zone of inhibition (area with no growth) will be seen around the disk after incubation if the organism is a Group A beta-hemolytic Streptococcus. Other beta-hemolytic streptococci are resistant to (not killed by) bacitracin. Their colonies will thus grow right up to the disk of bacitracin.


Reagents and Supplies:

  • Hardy Taxo A disks (0.04 IU Bacitracin)
  • 5% Sheep Blood  w/TSA plates
  • Sterile inoculating loops
  • 37 C Incubator


Reagent Storage

  • On receipt, store at -20 to +8°C. After use, store vial or cartridge to protect product integrity at 2 to 8°C.
  • Use oldest discs first and discard expired discs. Allow containers to come to room temperature before opening.
  • Return unused discs to the refrigerator when application of discs has been completed.
  • Vials and cartridges from which discs have been frequently removed during one week and discs left out overnight in the laboratory should be discarded, or the discs should be tested for performance with control organisms prior to continued use.

 Quality Control:

 QC is performed weekly by inoculating one half of a blood agar plate with S pyogenes ATCC 19615 and the other half with S agalactiae ATCC 12386. After 24 hr incubation the S pyogenes should show a distinct zone of inhibition while the S agalactiae should not.

Record result in Bacitracin test QC log.



  1. With a sterile inoculating loop, obtain a portion of an isolated colony of the strepcococci being tested.
  2. Streak the plate for isolation.
  3. With sterile forceps, obtain a bacitracin disk and place inon the inoculated agar at the intersection of the primary and secondary streaks. Tap the disk with forceps to ensure adherence to the agar surface.
  4. Incubate the blood agar plate in ambient air at 35°C for 18 to 24 hours.
  5. After the incubation period, examine the blood agar for a zone of growth inhibition around the bacitracin disk.


Interpretation:     Susceptible= any zone of growth inhibition

                             Resistant=no zone of growth inhibition            



HardyDisk Bacitracin Differentation Disks package insert 3/31/2016

Levinson, M.L and P.F. Frank 1955 Differentiation of Group A from other beta-hemolytic streptococci with bacitracin. J. Bacteriol. 69:284-287


Effective Date: 
Fri, 08/09/2013
Tue, 02/04/2020


Purpose and Scope:

The HardyDisk™ Novobiocin Differentiation Disks are useful in presumptively distinguishing S. saprophyticus from other coagulase-negataive Staphlococci. Other human staphylococcal species that are novobiocin-resistant (S. cohnii, S. xylosus, S. pulvereri) are rarely isolated from patients.

Coagulase-negative staphylococci  are among those organisms that have traditionally been considered skin contaminants, and their recovery from cultures doesn't always indicate presence of disease. Therefore, little attention had been paid to the pathogenic potential of this group of bacteria until recently. By the mid-1970's, microbiologists were becoming aware that coagulase-negataive staphlococci could indeed be pathogenic, especially in compromised hosts.

 Today, S. saprophyticus has proven to be an important uropathogen. It is second only to E. coli as the most common cause of cystitis and acute urinary tract infection (UTI) in healthy, young adult women. S. saprophyticus tends to adhere to uroepithelial cells more often and more successfully than other staphylococcal species, this is believed to partially explain the organism's frequent role in urinary tract infections.


Reagents and Supplies:

  • HardyDisk™ Novobiocin Differentiation Disks 5ug
  • Blood Agar plates
  • BD  BBL Prompt Inoculation System


Reagent Storage

  • Upon receipt store at -20 to +8 degrees C. away from direct light.
  • The disks should not be used if there are any signs of deterioration, discoloration, or if the expiration date has passed.
  • Protect from light, excessive heat, and moisture.
  • The expiration date applies to the product in its intact packaging when stored as directed.


Quality Control:


QC is performed on each day the test is performed. Use procedure with stock culture of Staph saprophyticus ATCC 15305  (Novobiocin resistant) and Staph epidermidis ATCC 12228  (Novobiocin sensitive).

Record QC results on Novobiocin Test QC Log.



  1. Allow disks to equilibrate to room temperature.
  2. Using a pure 18-24 hour culture, prepare a suspension using the Prompt inoculation system. (See Kirby Bauer Sensitivities Procedure SOP Micro-06 for instructions on using Prompt Inoculation system.)
  3. Use one or two colonies to inoculate a BAP.  Streak for isolation or just use a primary streak on the plate surface.
  4. Aseptically apply one novobiocin disk onto the primary streak of the inoculated agar surface and lightly press down to ensure full contact with the medium.
  5. Incubate aerobically for 18-24 hours at 35-37 degrees C.
  6. Measure (in millimeters) the diameter of the zone of inhibition around the novobiocin disk, and record as susceptible or resistant.

Sensitive - A zone of inhibition greater than 16mm.
Resistant - A zone of inhibition less than or equal to 16mm.



It is recommended that biochemical and/or serological tests be performed on colonies from pure culture for complete identification.

The novobiocin disk is not helpful and can give misleading results if it is performed on isolates other that those from urinary specimens.

Occasional human isolates that are not S. saprophyticus, S. cohnii subsp., or S. xylosis may also be resistant to novobiocin.








HardyDisk™ Novobiocin Differentiation Disks Package Insert Copyright 2013

Murray, P.R., et al. 2003. Manual of Clinical Microbiology, 8th ed. American Society for Microbiology, Washington, D.C.

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