IVATECT STREPTO A (20 Tests) Reference : IVN 25410 Insert version : 03-2003 For Qualitative Detection of Group A Streptococcal Antigen from Throat Swabs or Confirmation of Presumptive Group A Streptococcal Colonies from Culture I-SUMMARY AND EXPLANATION Among the beta-hemolytic streptococci causing infections in humans, the A, B, C and G groups figure most prominently (5). Group A streptococci continue to be a focus of interest not only because of their causal role in acute streptococcal pharyngitis and other pyogenic infections but also because of their association with post streptococcal sequelae, specifically acute rheumatic fever and acute glomerulonephritis (1,2). In order to properly treat the disease using antibiotic therapy, it is important to use an accurate diagnostic method to identify the pathologic agent. For the screening of group A streptococcal infection several methods are currently used including susceptibility of the organism to a bacitracin disc placed on a sheep blood agar plate, latex agglutination and enzyme immunoassay (3,4). II-PRINCIPLE The IVATECT STREPTO A test is a rapid qualitative one step assay for the detection of Streptococcus group A antigen from Throat swabs specimens. The method employs a unique combination of monoclonal-dye conjugate and polyclonal solid phase antibodies to selectively identify streptococcus A with a high degree of sensitivity. As the test samples flows through the absorbent pad, the labelled antibody-dye conjugate binds to the Strep-A antigen forming an antibody antigen complex. This complex binds to the Anti-Strep-A antibody in the test zone producing a pink-rose color band. In the absence of Strep-A, there is no line in the test zone. The reaction mixture continues flowing through the solid phase. Unbound conjugate binds to the reagents in the control zone producing a pink-rose color band, demonstrating that the reagents are functioning correctly. VIII-INTERPRETATION OF RESULTS B B- Positive : In addition to the control band, a clearly distinguishable band will appear near to the “bottom” indicating presence of Strep A in the sample. C- Inconclusive : The test is inconclusive in the absence of distinct band at the top and bottom areas of the reaction strip. It is recommended that in this case the test should be repeated. IX-PERFORMANCE DATA 1. ACCURACY A study was performed using a total of 375 patients. Samples were collected on duplicate throat swabs from patients complaining of a save throat. One of the swabs was tested using the IVATECT STREPTO A and the other swab was cultured on sheep blood agar plates containing a bacitracin disc. After 24 hours of incubation, all samples showing a zone of inhibition around the bacitracin disc were considered as positive. III- IVATECT STREPTO A COMPONENTS Each kit contains everything needed to perform 20 tests : - 20 Strep A Strip tests - 1 bottle of extraction solution 1 (6.5 ml) - 1 bottle of extraction solution 2 (6.5 ml) - Swabs are available optionnaly A A- Negative : Only 1 couloured band near to the “top” (control band) will appear. IVATECT STREPTO A CULTURE + - + - 36 2 1 336 IV-STORAGE AND STABILITY SENSITIVITY 36/38 94,7 % Strep A has to be stored at room temperature (4°C to 30°C) in the sealed pouch. Do not freeze the test kit. SPECIFICITY 336/337 99,7 % AGREEMENT 372/375 99,2 % V-PRECAUTIONS 1. 2. 3. 4. This test is designed for “IN VITRO” diagnostic use only Extraction reagents are caustic and may cause irritation to skin, eyes and mucus membranes. Wash off immediatly if extraction reagent came in contact with skin. Read carefully instruction notice using this test. Do not use beyond expiry date which appears in the package label. 2. SPECIFICITY Different strains of bacteria were tested using the IVATECT STREPTO A in order to determinate if they may cause non specific reaction. The following panel of organisms was obtained from the Center for Disease Control (CDC) and the American type Culture Collection (ATCC). SPECIES VI-SPECIMEN COLLECTION To obtain the best results, specimens should be collected using standard throat swabs collection methods. Plastic shafted swabs with rayon or dacron tips may be used. Do not use swabs with cotton or calcium alginate tips, with wooden shafts or impregnated with charcoal or transport media containing agar or gelatin. Patient samples are best performed immediatly following specimen collection. If immediate testing is not possible, the patient samples should be placed in a dry plastic tube and stored refrigerated at 2-8°C. VII-ASSAY PROCEDURE 1. 2. 3. 4. 5. Remove the strip test from the aluminium pouch. Place the specimen swab in a plastic tube (12 x 55 mm or comparable). Add 6 drops of extraction reagent 1 (300 µl) and 6 drops of extraction reagent 2 (300 µl). Twirl swab to mix the extraction reagents thoroughly. Incubate at room temperature for 2 mn minimum and 5 mn maximum. At the end of the incubation time, squeeze the swab firmly against the side of the tube in order to remove as such as possible liquid from the swab. Discard the swab. Dip the reaction strip into the tube containing the extract solution. Do not dip over the blue mark. The reaction strip can be left vertically until the end of the reaction time. Read results of the test between 5 to 10 mn after dipping. Streptococcus group A Streptococcus group B Streptococcus group C Ungrouped Streptococcus S. mutans S. epidermidis STRAIN RESULT CDC-799 CDC-1073 CDC-660 CDC-763 ATCC-27351 ATCC-155 Positive Negative Negative Negative Negative Negative X-LIMITATION OF THE TEST 1- IVATECT STREPTO A is a screening test for the presence of Group A streptococci. 2-As it is true with any diagnostic procedure, the physician should evaluate data obtained by the use of this kit in light of other clinical information, including culture, if results are inconsistent with clinical presentation. 3-This test will not differenciate between colonized and infected individuals. 4-Pharyngitis can be caused by organisms other than Group A streptococci XI-BIBLIOGRAPHY 1. Appleton, R.S., B.E. Victoria, D.Tamer, and E.M. Ayoub, J.Lab clin. Med, 105 : 114-119,1985. 2. Ayoub, E.M. and L.W. Wannamaker, Pediatrics, 29 : 527-538, 1962. 3. Bisuo, A.L. and I Ofek, J.dis; Child, 127 : 676-681,1974. 4. Levinson, M.L. and P.F. Franck, J.Bacteriol, 69 : 284-287, 1955. Manufactured by : IVAgen S.A. Parc Scientifique et Technique G. Besse Espace Innovation 2 110, allée C. Babbage 30035 NIMES Cedex FRANCE