Procedure: Whole Blood anti-HIV Antibody by OraQuickPRINCIPLECLINICAL SIGNIFICANCETEST PROCEDURE: EXTERNAL QUALITY CONTROL SOLUTIONSTEST PROCEDUREU M D N J _ R O B E R T W . J O H N S O N M E D I C A L S C H O O L P O C T P R O G R A MWhole Blood HIV-1 Antibody by OraQuickC H A P T E R 1 0 : Q U A L I T A T I V E H I V - 1 S C R E E N I N G B Y O R A Q U I C K1 of 7Chapter10POCT Program PROCEDUREProcedure: Whole Blood anti-HIV Antibody by OraQuickPRINCIPLEThe OraQuick Rapid HIV-1 Antibody Test is a single-use, qualitative immunoassay to detect antibodies to Human Immunodeficiency Virus Type 1 (HIV-1) in fingerstick whole blood specimens. The OraQuick is a manually performed, visually read, 20-40 minute immunoassay system. It is comprised of a single-use test device and a dingle use vial containing a pre-measured amount of a buffered developer solution. The OraQuick test utilizes a lateral flow procedure. The assay test strip contains synthetic peptides representing the HIV envelope regionin the Test zone, and a goat anti-human IgG procedural control in the Control zone, immobilized onto a nitrocellulose membrane.A fingerstick whole blood specimen is collected and transferred into a vial of developer solution, followed by insertion of the test device into the developer vial. The developer solution facilitates the flow of the specimen into the device and onto the test strip. As the diluted specimen flows through the devices, it rehydrates the protein-A gold colorimetric reagent contained in the device. As the specimen continues to migrate up the strip, it encounters the T zone. If the specimen contains antibodies that react with the antigens immobilized on the nitrocellulose membrane, a reddish-purple line will appear, indicating the presence of antibodies to HIV-1 in thespecimen. Further up the strip, the sample will encounter the C zone. A reddish-purple line at the C zone serves to demonstrate that a specimen containing antibodies was added to the vial and that the fluid has migrated adequately through the test device.CLINICAL SIGNIFICANCEAcquired Immune Deficiency Syndrome (AIDS), Aids related complex (ARC) and pre-AIDS are thought to be caused by the Human Immunodeficiency Virus (HIV). The first AIDS-related virus, HIV-1, has been isolated from patients with AIDS, and from healthy persons at high risk for AIDS. To date, eight HIV-1 subtypes have been identified world-wide, in addition to the highly divergent HIV-1 isolates form AIDS patients in Cameroon. A closely related but distinct second virus, designated HIV-2, has been isolated from West African patients with AIDS. Despite sharing a number of conserved genetic sequences, serological cross-reactivity between HIV-1 and HIV-2 is very variable from sample to sample. Using a rapid HIV test increased the number of HIV-infected persons who may be diagnoses. the CDC estimates that nearly one third of the estimated 900,000 HIV-infected persons in the United States do not know their HIV status. They cannot benefit from early intervention with effective antiviral therapy. The OraQuick test provides a rapid preliminary result and is suitable for use in multi-test algorithms designed for statistical validation of rapid HIV test results.SPECIMEN REQUIREMENTSNo special patient preparation is needed. However, all test subjects should have received the “Subject Information” pamphlet prior to specimen collection.A fingerstick blood sample should be collected using antiseptic technique. The sample must be collected immediately with the collection loop provided in the test kit, and must be tested immediately.REAGENTSOraQuick® Rapid HIV-1 Antibody Test Kit -- Catalog #: 1001-0052 (25 tests/kit), orOraQuick® Rapid HIV-1 Antibody Test Kit -- Catalog #: 1001-0051 (100 tests/kit)Vendor: OraSure Technologies, Inc., Bethlehem, PA, 800-672-7873REAGENTS ARE TO BE OBTAINED BY CONTACTING RWJMS POCT PROGRAM (SEE CONTACT LIST)E A C H K I T C O N T A I N S- 25 (or 100) Divided pouches, each containing a test device, developer solution vial and absorbent packet- 5 (or 10) Reusable test stands- 25 (or 100) Specimen collection loops- 25 (or 100) Subject information pamphletsM A T E R I A L S R E Q U I R E D B U T N O T P R O V I D E D :• Timer or watch for timing 20-40 minutes• Latex, vinyl or nitrile disposable gloves• Biohazard waste container• Clean, disposable, absorbent workspace coverThe test devices are stable until the expiration date on the box and pouch. They should be stored at room temperature, 35-80°F. If stored refrigerated, the pouch must be brought to room temperature before opening.QUALITY CONTROLEXTERNAL QUALITY CONTROL:OraQuick® Rapid HIV-1 Antibody Test Kit Controls – Catalog #:1001-0049Vendor: OraSure Technologies, Inc., Bethlehem, PA, 800-672-7873E A C H K I T C O N T A I N S- Positive Control, 1 vial (Black Cap, 0.2 mL)- Negative Control, 1 vial (White cap, 0.2 mL)The controls are human, plasma-based reagents. The positive control contains antibodies that will show a REACTIVE result and the Negative Control will show a Non-Reactive result when run with the Orasure Rapid HIV-1 Antibody Test. The controls are liquid, ready to use and require no reconstitution or dilution. Both controls contain .2 mL of photochemically inactivated human plasma which is either positive or negative for antibodies to HIV-1 diluted by a defibrinated pool of normal human plasma. Both specimens are negative for Hepatitis B surface antigen and Hepatitis C antibody.The controls should be stored at room temperature (2 - 8-C) and are stable until the expiration date stated on the the box label. HOWEVER, ONCE OPENED THE CONTROLS EXPIRE IN 21 DAYS. This expiration date should be clearly marked on the box containing the controls or on the individual vialsExternal quality control is to be performed:- Whenever a new shipment of reagents is put into use, both positive and negative controls shall be run on the first new box used from that shipment.- At the beginning of each week, at least one control must be run, to make sure that the reagents have not deteriorated in storage.- Each month, each person doing testing must run at least one control as a means of demonstrating continued competency.- The quality control requirements listed above can overlap:1. If more than one person will be doing testing, they may divide up the quality control, with one person running the negative