STREP A

VCUHSMCV Hospital and PhysiciansPoint of Care TestingPage 1 of 6SUBJECT: STREP A: BIOSTAR ACCEAVA STREP A Original Date: 3/1999IMMUNOASSAY TEST FOR THE QUALITATIVE Effective Date: 9/2007DETECTION OF GROUP A STREPTOCOCCUSI. PURPOSE:1. The Biostar Acceava Strep A Test allows for the rapid detection of Group A streptococcal antigendirectly from patient throat swab specimens. The test is intended for use as an aid in the diagnosis ofgroup A streptococcal infection.2. Group A Streptococci are organisms that typically cause illnesses such as tonsillitis, pharyngitis andscarlet fever. These infections can lead to serious complications, including rheumatic fever and acuteglomerulonephritis. Rapid diagnosis and appropriate antibiotic therapy of Group A streptococcalinfections appear to be the best means of preventing these complications.3. Group A Streptococci are responsible for about 19% of all upper respiratory tract infections, but theincidence varies by clinical setting. Streptococcal pharyngitis is seasonal in nature with the highestprevalence found during the winter and early spring. The highest incidence of this disease is found incrowded populations such as military bases and in school-aged children, and is evenly distributedbetween males and females.II.DEFINITION:The traditional means of detecting group A streptococcal infection involves 24-48 hour culture of throatswab specimens or other exudates. It is also possible to identify Group A Streptococci by immunologicalmeans. The cell walls of Group A Streptococci share a common, group-specific, antigenic determinant.This determinant is a carbohydrate consisting of polyrhamnose chains terminating in N-acetyl glucosamineresidues. Antibodies against this carbohydrate antigen have been used in a variety of tests to identify thegroup A organism.The Acceava Strep A Test uses color immunochromatographic dipstick technology with rabbit antibodiescoated on the nitrocellouse membrane. In the test procedure, a throat swab is subjected to a chemicalextraction of carbohydrate antigen unique to Group A Streptococcus. The Test Device is then placed in theextraction mixture and the mixture migrates along the membrane. If Group A Streptococcus is present inthe sample, it will form a complex with the anti-Group A Streptococcus antibody conjugated colorparticles. The complex will be bound to the anti-Group A Streptococcus capture antibody and a visible blueTest line will appear to indicate a positive result.A red control line will also be visible on the strip to validate the performance of each individual test device.A proprietary protein at the bottom of the strip migrates up the strip with the patient sample. Once thispropriety antigen reacts with the antibody embedded on the designed portion of the test device thechromagen is activated and a red line appears. If there is not enough sample or if the sample fails tomigrate, there will be no control line. Also if there is an interfering substance, which binds the proprietyantigen, there will be no control line.III.POLICY:A. GENERAL1. Staff who has documented evidence of training and competency may perform Strep A testing.2. New personnel will be oriented to the procedures for performing Strep A testing duringorientation.

VCUHSMCV Hospital and PhysiciansPoint of Care TestingPage 2 of 63. Skill validation in the testing procedure will occur annually. Individual records documentingcurrent satisfactory performance will be maintained in each testing area.4. Patient specimens, and all materials that come in contact with them, should be handled aspotentially infectious and disposed of according to Standard Precautions guidelines.B. SPECIMEN1. Throat swab specimens must be collected by standard clinical methods using the sterile rayonswabs supplied with the kit.2. Use only rayon tipped swabs with solid plastic shaft or dacron swabs to collect the specimen. Donot use swabs that have cotton tips or wooden shafts. Do not use calcium alginate swabs.3. Process specimens as soon as possible after collection. The swabs can be held in any clean, dryplastic tube or sleeve for up to 24 hours if refrigerated (2-8 C) before processing. The swabs andthe test kit must be at room temperature before performing test. Performance with transport mediahas not been assessed. Modified Stuart’s Transport Media in an acceptable liquid media. The useof charcoal or semisolid transport media collection systems not recommended.C. REAGENT AND MATERIALSBiostar Acceava Strep A Kit• Catalog # ACCGAS (50-test)• Store kit at room temperature, 58-86 F (15-30 C), out of direct sunlight. Kit contents arestable until the expiration date printed on the outer box. Do not use beyond the Expirationdate printed on the outside of the box. Do not freeze.Kit contains:1. 50 Test Devices-Rabbit polyclonal antibody to strep A (Test Line) and a proprietary protein antigen at the bottomof the device, which migrates up the strip and binds to the antibody embedded on the strip(Control Line).-The Test Cassette must remain sealed in the protective foil pouch until just prior to use2. 50 Test Tubes-Where extraction of Strep antigen from the swab takes place.3. 50 individually packaged sterile rayon-tipped swabs on solid shafts4. Reagent 1 (2 M Sodium Nitrate) Caution: Toxic5. Reagent 2 (0.3 M Acetic Acid) Caution: CorrosiveIf Reagent 2 comes into contact with skin or eyes, flush with large volumes of water.6. 1 Positive Control solutionNonviable Group A Streptococci, 0.1% Sodium Azide*7. 1 Negative Control solutionNonviable Group C Streptococci, 0.1% Sodium Azide*

VCUHSMCV Hospital and PhysiciansPoint of Care TestingPage 4 of 6being transferred to the patient chart. Once the result is transferred to the chart, the persontranscribing the result should reconfirm the result. If a patient test is performed by anyone otherthan a licensed professional, the test result must be initialed by a licensed professional when it istransferred to the patient chart.3. If an error is detected by a licensed professional upon review of the test result, corrective actionsmust be initiated immediately. The corrective action involves making the correction to the patientchart and initialing it.4. Point-of-care testing results that do not correlate with patient condition should be followed up byconfirmatory testing in the main laboratory.5. Strep A Logs will be reviewed weekly by the Nurse Manager or designee and monthly bylaboratory personnel. Weekly/Monthly review is documented on the Strep A Log.IV.PROCEDURES:PROCEDURE A: PATIENT TESTING/DAILY QUALITY CONTROLRequisites:Biostar Acceava Test DeviceBiostar Acceava Test TubesBiostar Acceava Reagent 1 (2M Sodium Nitrite)Biostar Reagent 2 (0.3 M Acetic Acid)Biostar Acceava Rayon tipped swabTongue Blade or SpoonGlovesTiming DeviceTesting Procedure:1. Verify patient identification and explain the procedure to patient.2. Wash hands with soap and friction for 15 seconds.3. Put on gloves.4. Using a rayon-tipped swab provided in the kit, collect a throat swab specimen. Depress the tongue with a tongueblade or spoon. Be careful not to touch the tongue, sides or top of the mouth with the swab. Rub the swab on theback of the throat, on the tonsils, and in any other area where there is redness, inflammation or pus.5. Remove a Test Tube from the kit and place it upright in the plastic rack provided by the kit.6. Just before testing, add four drops Reagent 1 (pink) and four drops Reagent 2 to the test tube. With the additionof Reagent 2 the solution should turn from pink to clear.7. Immediately place the swab in the tube. Vigorously mix the solution by rotating the swab forcefully against theside of the plastic tube ten times. Best results are obtained when the specimen is vigorously extracted in thesolution.8. Let the swab stand in the solution for one minute.9. After one minute express as much liquid as possible from the swab by squeezing the sides of the tube as theswab is withdrawn.10. Discard the swab.

VCUHSMCV Hospital and PhysiciansPoint of Care TestingPage 5 of 611. Remove a Test Device from the container, and recap the container immediately.12. Place the absorbent end of the Test Device into the extracted sample.13. Read results at 5 minutes. Some positive results may be seen earlier.Interpretation of Results:Positive Result – A blue or red line that appears uneven in color density is considered a valid result. In cases ofmoderate or high positive specimens, some blue color behind the line may be seen; as long as the Test Line andControl Line are visible results are valid.Negative Result – The appearance of only the red Control Line but no blue Test Line is a presumptive negativeresult.Invalid Result – If after ten minutes, no red Control Line appears or background color makes it difficult to readthe red Control Line, the result is invalid. If this occurs, repeat the test on a new stick.14. Document all patient test results, qualify control results (Control Line), and quality control corrective actions onthe Strep A Log.15. Dispose of patient specimens and all materials that come in contact with them according to StandardPrecautions guidelines.PROCEDURE B: QUALITY CONTROL TESTING (When Opening A New Kit)Requisites:Acceava Positive Control SwabAcceava Negative Control SwabAcceava Strep A Test CassetteAcceava Strep A Extraction Solution BottleGlovesTiming DeviceQuality Control:1. Wash hands and put on gloves.2. Remove two plastic tubes from kit and place upright in the plastic rack provided in the kit.3. Dispense four drops of Reagent 1 and four drops of Reagent 2 into test tubes.4. Vigorously mix the control contents. Add 1 free falling drop of Control from the dropper bottle.5. Place a clean swab into the test tube.6. Vigorously mix the solution by rotating the swab forcibly against the side of the tube at least ten times. Bestresults are obtained when the specimen is vigorously extracted in the solution.7. Let stand for one minute.8. Express as much of the liquid as possible from the swab by squeezing the sides of the Tube as the swab iswithdrawn.9. Discard the swab.

VCUHSMCV Hospital and PhysiciansPoint of Care TestingPage 6 of 610. Remove Test Devices from the container; re-cap container immediately.11. Place the Absorbent end of the Test Device into the Extracted Sample.12. Read results at 5 minutes (some positives may be seen earlier).13. See Procedure A: Patient Testing – Interpretation of Results for interpretation. If quality controls do notperform as expected:Do not use the kit for patient testing. Consult the Charge Nurse, Clinical Coordinator, Nurse Manager, or Pointof-CareTesting Supervisor.14. Document quality control results and quality control corrective actions on the Strep A Log.15. Dispose of all materials according to Standard Precautions guidelines.APPENDIX B: ABOUT THE TESTReference Range:NegativeProcedural Limitations Provided by Manufacturer:1. The Biostar Acceava Strep A Test has been categorized as CLIA waived only for the application of qualitativedetection of Group A Streptococcal Antigen from throat swabs. The application for the confirmation ofpresumptive Group A Streptococcal colonies recovered from culture is not waived.2. Respiratory infections, including pharyngitis, can be caused by Streptococcus from serogroups other than groupA as well as other pathogens. The Biostar Acceava Strep A Test will not differentiate asymptomatic carriers ofGroup A Streptococcus from those exhibiting streptococcal infection.3. Test results must always be evaluated with other data available to the physician. A negative test result occurs ifthe level of extracted antigen in a sample is below the sensitivity of the test or if the specimen is inadequate.Additional follow-up testing using the culture method is recommended if the Biostar Acceava Strep A test resultis negative. This is the recommendation of the American Association of Pediatrics as referenced in the PediatricRedbook (1994).4. The Acceava Strep A test should only be used with throat swabs, or colonies taken directly from a plate. Theuse of swab specimens taken from other sites or the use of other samples such as saliva, sputum, or urine as notbeen established. The quality of the test depends on the quality of the sample: proper throat swab specimensmust be obtained.V. REFERENCES:Biostar Acceava Strep A Test Package Insert. Biostar, Inc., Boulder CO 2006.VI.RESOURCES:Department of Clinical Pathology