QuickVue Dipstick Strep A Test PDF

28 INTENDED USE The QuickVue Dipstick Strep A Test is intended for the rapid, qualitative detection of Group A Streptococcal antigen from throat swabs or confirmation of presumptive Group A Streptococcal colonies recovered from culture. The test is to be used to aid in the diagnosis of Group...

28 INTENDED USE The QuickVue Dipstick Strep A Test is intended for the rapid, qualitative detection of Group A Streptococcal antigen from throat swabs or confirmation of presumptive Group A Streptococcal colonies recovered from culture. The test is to be used to aid in the diagnosis of Group A Streptococcal infection. For use by healthcareprofessionals. SUMMARY AND EXPLANATION Group A Streptococcus is one of the most important causes of acute upper respiratory tract infection. Early diagnosis and treatment of Group A Streptococcal pharyngitis has been shown to reduce the severity of symptoms and serious complications such as rheumatic fever and glomerulonephritis.1 Conventional identification procedures for Group A Streptococcus from throat swabs involve the isolation and subsequentidentification of viable pathogen techniques that require 24 to 48 hours or longer for results.2 PRINCIPLE OF THE TEST The QuickVue Dipstick Strep A Test is a lateral-flow immunoassay utilizing Quidel’s patented antibody- labeledparticles. The test detects either viable or nonviable organisms directly from throat swabs or culture colonies within 5 minutes. To perform the test, a Throat Swab specimen is collected. Antigen is extracted from the Swab specimen withReagents A and B. The Dipstick is then added to the extracted sample. If the sample contains Strep A antigen, a pink-to-red Test Line along with a blue procedural Control Line willappear on the Dipstick, indicating a positive result. If Strep A antigen is not present, or present at very low levels, only a blue procedural Control Line will appear. REAGENTS AND MATERIALS SUPPLIED Each kit contains: ◼ Individually packaged Dipsticks (25 or 50): Dipsticks coated with rabbit polyclonal anti- Group AStreptococcus ◼ Extraction Reagent A (1): Contains 4 M sodium nitrite ◼ Extraction Reagent B (1): Contains 0.2 M acetic acid ◼ Sterile Throat Swabs (25 or 50) ◼ Tubes (25 or 50) ◼ Positive Control (1): Heat-inactivated Group A Streptococcus with 0.2% sodium azide ◼ Negative Control (1): Heat-inactivated Group C Streptococcus with 0.2% sodium azide ◼ Package Insert (1) ◼ Procedure Card (1)  29 WARNINGS AND PRECAUTIONS ◼ For in vitro diagnostic use ◼ Do not use beyond the expiration date printed on the outside of the box. ◼ The Dipstick must remain sealed in the protective foil pouch until just prior to use. ◼ Reagent B contains an acidic solution. If the solution contacts the skin or eye, flush with large volumes ofwater. ◼ Use of nitrile or latex gloves is recommended when handling the Extraction Reagents within this kit.3,4 ◼ Do not interchange reagent bottle caps. ◼ If Reagent B is green prior to mixing with Reagent A in the Tube, do not use and contact Technical Support. ◼ To obtain accurate results, you must follow the Package Insert instructions. ◼ Testing should be performed in an area with adequate ventilation. ◼ Dispose of containers and unused contents in accordance with Federal, State and Local regulatoryrequirements. ◼ Wear suitable protective clothing, gloves, and eye/face protection when handling the contents of this kit. ◼ Wash hands thoroughly after handling. ◼ For additional information on hazard symbols, safety, handling and disposal of the components within thiskit, please refer to the Safety Data Sheet (SDS) located at quidel.com. KIT STORAGE AND STABILITY Store the kit at room temperature, 59°F to 86°F (15°C to 30°C), out of direct sunlight. Kit contents are stableuntil the expiration date printed on the outer box. Do not freeze. SPECIMEN COLLECTION AND STORAGE Collect throat swab specimens by standard clinical methods. Consult standard reference procedures such as the collection method described by Miller and Holmes.5 Depress the tongue with a tongue blade or spoon. When swabbing the throat, be careful not to touch the tongue, sides or top of the mouth with the Swab. Rubthe Swab on the back of the throat, on the tonsils and in any other area where there is redness, inflammationor pus. Use rayon tipped swabs to collect throat specimens. Do not use calcium alginate, cotton tipped or woodenshaft swabs. It is recommended that swab specimens be processed as soon as possible after collection. Swabs can be held in any clean, dry plastic tube or sleeve up to 72 hours at room temperature (15°C to 30°C), or refrigerated (2°Cto 8°C) before processing. The use of charcoal or agar medium is not recommended. If a culture is desired, lightly streak the swab on a 5% sheep blood agar plate before using the Swab in the QuickVue Dipstick Strep A Test. Do not perform the QuickVue Dipstick Strep A Test before streaking the Swab,as the Extraction Solution will destroy the bacteria on the swab, thereby rendering the organism incapable of successful culturing. Alternatively, throat swab specimens can be obtained by dual Swabs or by two sequentialSwabs for the culture procedure. CULTURE CONFIRMATION The QuickVue test can be used to confirm the identification of Group A Streptococcus on blood agar plates.Lightly touch a colony using a sterile Swab. Do not sweep the plate. Follow the instructions in the TEST PROCEDURE section to test the Swabs.  30 QUALITY CONTROL Built-in Control Features The QuickVue Dipstick Strep A Test provides three levels of internal procedural controls with each test run. Fordaily quality control, Quidel recommends documenting that these internal controls were checked for the first sample tested each day. ◼ The color of the Extraction Reagent changes from clear to green as the reagents are mixed together. The color change is an internal extraction reagent control and is an indication that the reagents are mixed andfunctioning properly. ◼ The appearance of a blue Control Line is an internal control. The Dipstick must absorb the proper amountof sample and the Dipstick must be working properly for the blue Control Line to appear. Additionally, theappearance of the Control Line indicates that capillary flow occurred. ◼ A clear background is an internal background negative control. If no interfering substances are in the sample and the Dipstick is working properly, the background in the Result area should be white to lightpink within 5 minutes and not interfere with the reading of the test result. External Quality Control Testing External controls may also be used to demonstrate that the reagents and assay procedure perform properly. Quidel recommends that Positive and Negative Controls be run once for each untrained operator, once for each new shipment of kits - provided that each different lot received in the shipment is tested - and as deemedadditionally necessary by your internal quality control procedures, and in accordance with Local, State, and Federal regulations or accreditation requirements. If the controls do not perform as expected, repeat the test or contact Quidel Technical Support before testingpatient specimens. Normal Values: Negative ASSAY PROCEDURE ◼ Do not remove Dipsticks from the foil pouch until ready to perform the assay. ◼ To avoid cross contamination, do not allow the tip of the reagent bottles to come in contact with sampleSwabs.  31 TEST PROCEDURE Important: ◼ Gloves should be worn when handling samples. ◼ Do not use Reagent B if the solution is green prior to mixing with Reagent A in the Tube. If this occurs,contact Technical Support. 1. Just before testing, add 3 DROPS of Reagent A and 3 DROPS of Reagent B into aclean Tube. This solution should turn green. When adding drops, hold bottle vertically so that a complete drop forms. 2. Immediately add the patient Swab sample to the Tube. Squeeze the bottom of theTube so the Swab head is compressed. Rotate the Swab a minimum of 5 times. Keep swab in Tube for 1 minute. 3. Express all liquid from the Swab against the inside of the tube. Squeeze the Swabfirmly as it is removed from the Tube. Discard the Swab. 4. Remove the Dipstick from the foil pouch. Place the Dipstick into the Tube with the arrows of the Dipstick pointing down. Do not handle or move the Dipstick until the testis complete and ready for reading. 5. Read result at 5 minutes. Some positive results may appear sooner.  32 INTERPRETATION OF RESULTS AND REPORTING Positive Result: Positive Any pink to red Test Line along with any shade of a blue procedural Control Line is a positive result for thedetection of Group A Streptococcus antigen. Negative Result: Negative A blue procedural Control Line and no pink Test Line is a presumptive negative result. Invalid Result: The test result is invalid if a blue Control Line is not visible at 5 minutes. If this occurs, retest using a newsample and a new Dipstick or contact Technical Support. Normal Value: Negative QC TESTING PROCEDURE ◼ Follow the instruction procedures in the TEST PROCEDURE to dispense the Extraction Reagents into theTube (step 1). ◼ Vigorously mix the Control Bottles. Add 1 drop of the Negative or Positive Control into the Tube. ◼ Place a clean Swab into the Tube and follow the instructions for testing the patient Swab. LIMITATIONS The contents of this kit are for use in the qualitative detection of Group A Streptococcal antigen from throatswab specimens and culture colonies only. Failure to follow the test procedure and interpretation of test results may adversely affect performance and/or produce invalid results. The test detects both viable and nonviable Group A Streptococci and may yield a positive result in the absenceof living organisms. Respiratory infections, including pharyngitis, can be caused by Streptococcus from serogroups other than Group A as well as other pathogens. The QuickVue Dipstick Strep A Test will not differentiate asymptomatic carriers of Group A Streptococcus from those exhibiting Streptococcal infection.6 Some commercial controls may contain interfering additives and are not recommended for use in theQuickVue test. Test results must always be evaluated with other data available to the physician. A negative test result might occur if the level of extracted antigen in a sample is below the sensitivity of the test or if a poor- quality specimen is obtained. Additional follow-up testing using the culture method is recommended if the QuickVuetest result is negative.  33 Remel RPR CARD TEST INTENDED USE Remel RPR Card Test is a nontreponemal flocculation test intended for detection of reagin (anti-lipoidal antibodies) in human serum for presumptive serological diagnosis of syphilis when used in conjunction with a treponemal test. SUMMARY AND PRINCIPLE RPR Card Test is a nontreponemal test for serological detection of syphilis,recommended for use when venous blood is collected. RPR Antigen consists of antigens derived from sources not directly associated with Treponemal microorganisms1. 3 In this method, carbon-particle cardiolipin antigen detects reagin, a substance present in sera of syphilitic persons and occasionally in sera of persons with other acute or chronic conditions.4 Reagin is an antibody-like substance produced from the reaction of treponemal microorganisms with body tissue. The detection of reagin in the diagnosis of syphilis. A fourfold decrease in titer following syphilis treatment indicates that treatment has been successful; a fourfold increaseindicates either treatment failure or reinfection. In serum containing reagin, flocculation occurs with agglutination of the carbon particles in the RPR Antigen. Black clumps appear against a whitebackground which can be read macroscopically. In contrast, nonreactive specimens appear to have a uniform light-gray color. PRECAUTIONS This product is for /n Vitro diagnostic use and should be used by properly trained individuals. Precautions should be taken against the dangers of microbiological hazards by properly sterilizing specimens, containers, and test materials after use. Carefully read the entire procedure prior to performing any tests. 1. Potential Biohazardous Material: All human serum should be considered potentially infectious and handled accordingly. Refer to the current edition of Biosafety in Microbiological and Biomedical Laboratories for information on handling human specimens.8 2. RPR Antigen contains thimerosal as a preservative, which may be toxic if ingested. 3. Refer to Material Safety Data Sheet for detailed information on reagent chemicals. STORAGE Store product in its original container at 2-8°C until used. Once the kit is opened, store the RPR Antigen at 2- 8°C and all other kit components in a dry place at room temperature. Once the RPR Antigen is placed in the plastic dispensing bottle it is stable for 3 months or until the expiration date on the kit, provided it is stored at 2-8°C. Do not freeze or expose to temperature extremes. PRODUCT DETERIORATION This product should not be used if (1) the appearance of the reagents has changed, (2) there is evidence of contamination, (3) the expiration date has passed, or (4) there are other signs of deterioration. SPECIMEN COLLECTION AND HANDLING Use serum prepared from whole blood collected without anticoagulant. Allow blood to fully clot before centrifuging. Serum should be clear and separated from cells as soon after collection as possible. Hemolyzed specimens are not acceptable for testing when printed matter cannot be read through them.  34 MATERIALS AND REAGENTS PROVIDED Tests/Kit Components: 150 500 RPR Antigen: 0.003%Cardiolipin, 0.09% Cholesterol, 1 x3 ml 3x3 ml 0.021% Lecithin, 0.0125 M EDTA, 0.01 M Na2HP0 4, 0.01 M KH2P0 4, 0.01875% Charcoal, 0.1% Thimerosal (preservative), 10.0% Choline Chloride, w/v, Demineralized Water Plastic Dispensing Bottle 1 each 1 each 20 Gauge, Galvanized Needle, Blunt Cut 1 each 1 each White. 10 well Test Cards 15 each 50 each Pioette/Stirrers. 50 ul 150 each 500 each REF Number R16302 R16303 MATERIALS REQUIRED BUT NOT SUPPLIED (1) Mechanical rotator with fixed speed or adjusted to 100 rpm, circumscribing a ¾" circle, (2) Humidifier cover with moistened blotter or sponge, (3) 1 ml pipette, (4) High intensity incandescent lamp, (5) RPR Liquid Controls {REF R16307) or suitable alternative, (6) Calibrated pipette, 100 µI, 50 µI, (7) Test lubes (12 X 75), (8) Normal saline (0.85%), (9) Human serum, nonreactive for syphilis (for quantitative procedure). PROCEDURE Preparation of Reagents: Check the delivery of the needle by placing it firmly on a syringe. Fill the pipette with 1ml of water. Hold it in a vertical position and dispense 1.0 ml of water while simultaneously counting the drops. Aneedle delivery rate of 60 drops +/- 2 drops is acceptable. If the needle does not meet this specification it should not be used. Allow RPR Antigen to equilibrate to room temperature. Gently shake the bottle for 10-15 seconds to resuspend the antigen. Attach the needle (provided) to the tapered fitting on d i s p e n s i n g b o t t l e. Upon completion of the daily tests, remove the needle from the dispensing bottle and rinse with demineralized water. This will help maintain clear passage of the suspension. Do not wipe the needle as this may affect the accuracy of the antigen drop as it is dispensed. Allow RPR Antigen, RPR Liquid Controls, and test specimens to equilibrate to room temperature prior to use. The temperature of the room and all test materials, including specimens, must be maintained in the range of 23-29°C. Temperatures below this range will cause false-negative reactions and lower titers, while temperatures above the range have the opposite effect.7 Verify the speed of the mechanical rotator (100 +/- 2 rpm) to ensure reproducible results. Handle test cards by grasping the edge of the card; do not touch the surface of the test wells. RPR controls with established patterns of reactivity should be included in each test run. RPR Liquid Controls are available under a separate reference number (R16307).  35 Qualitative Test Procedure: 1. Label the test wells on the card with the specimen identification 2. Use a separate pipette/stirrer for each test specimen or control. Pre- squeeze the pipette/stirrer and draw up the specimen or control. Dispense 1 free-falling drop (50 µI) into the appropriate well. 3. Using the opposite, flattened end of the pipette/stirrer gently spread the specimen or control over the entire circle using a circular motion. 4. Gently shake RPR Antigen suspension in the dispensing bottle. Holding the bottle in a vertical position, dispense several drops into the cap to verify the needle passage is clear. Dispense 1 free-falling drop into each well containing specimen or control. Do not stir; mixing of the antigen suspension and the sample is accomplished during rotation. 5. Immediately place the test card on the mechanical rotator, cover withthe humidifier cover, and rotate for 8 minutes at 100 rpm. Note: False-positive reactions may occur due to evaporation if samples are not properly covered during rotation. 6. Following the 8-minute rotation, briefly rotate and tilt the card back and forth by hand 3-4 x to aid in differentiating nonreactive from minimally reactive results. Immediately read the card macroscopically in the wet state under a high intensity incandescent lamp. Avoid glare when reading reactions. Reading Report Small to large clumps (R) or slight but definite clumps (Rm) Reactive (R) No clumping or very slight roughness Nonreactive (NR) Note: All specimens with a reactive or rough reaction in the Qualitative Test should be tested according to the Quantitative Test procedure to provide a baseline from which changes can be determined, particularly for evaluating the efficiency of treatment\. Initial reports should only be made on specimens that are nonreactive. QUALITY CONTROL RPR controls with established patterns of reactivity should be included in each test run. A test run can be defined as a period of approximately 24 hours. Use reactive, minimally reactive, and nonreactive controls and test according to the Qualitative Test procedure. Remel RPR Liquid Controls can be obtained under a separate reference number (REF R16307). Each laboratory should establish endpoint titers for the quantitative controls used. If controls do not perform as expected, patient results should not be reported. Quality control testing should be performed according to established laboratory quality control procedures following the guidelines and recommendations of applicable federal, state, and local regulatory agencies. LIMITATIONS OF PROCEDURE 1. A diagnosis of syphilis should not be based on a single reactive serologic test. All historical information, clinical findings, and laboratory results should be taken into consideration. · 2. Biologic false-positive (BFP) reactions occurring with nontreponemal tests may be acute or chronic. Acute false- positives (titers lasting 6 months) have been observed with connective tissue diseases (e.g., systemic lupus erythematosus), narcotic addiction, aging, leprosy, and malignancy. Unusually high false-positive titers may be seen in patients diagnosed with lymphoma. 3. The RPR test cannot be used to test cerebrospinal fluid.