RPR TEST KITS. Newmarket Laboratories Ltd Lanwades Business Park, Kentford, Newmarket, CB8 7PN UK

Transcription

1 Product Code: RPR TEST KITS Kits for the qualitative and semi-quantitative detection of syphilis antibody in human serum or plasma by macroscopic agglutination on disposable test cards. Newmarket Laboratories Ltd Lanwades Business Park, Kentford, Newmarket, CB8 7PN UK Distributed by: Bio-Rad 3 Boulevard Raymond Poincaré Marnes - la - coquette - France

2 RPR 100 RPR 500. Code : 100 tests Code : 500 tests Kits for the qualitative and semi-quantitative detection of syphilis antibody in human serum or plasma by macroscopic agglutination on disposable test cards. All manufactured and commercialised reagents are under complete quality system starting from reception of raw material to the final commercialisation of the product. Each lot is submitted to a quality control and is only released when conforming to the acceptance criteria. The records relating to production and control of each single lot are kept within our company. Clinical Background. Syphilis is a chronic infection that progresses through distinct stages of infection: primary, secondary, tertiary, and quaternary. These stages produce diverse clinical symptoms, typically producing initial chancres then syphilitic rash followed by long periods of dormancy. Untreated infection may eventually result in cardiovascular problems and neurosyphilis. Caused by the spirochaete Treponema pallidum, infection is usually acquired by sexual contact, although the disease may be transmitted by transfusion of infected blood. Intra-uterine infection also occurs. The organism has proved virtually impossible to culture in artificial media, and diagnosis of the infection usually depends on the demonstration of antibodies in the blood, which appear soon after initial infection, and may persist for many years. The RPR is a non-treponemal test, in that the antibodies detected are not specific for T pallidum, although their presence in patient s serum or plasma is strongly associated with infection by the organism. This type of test measures antibody (IgG and IgM) produced in response to lipoidal material released from damaged host cells as well as to lipoprotein-like material released from the spirochaetes. These antibodies tend to disappear after successful cure of the infection. Intended Use These kits are intended for use by appropriately trained and qualified personnel for the detection of syphilis-associated antibodies in human serum and plasma. Principle of the test The RPR 100 & 500 kits use carbon particles coated with a mixture of lipid antigens, which will combine with antibody present in patient s serum or plasma. The particles are suspended in a medium containing components to eliminate non - specific reactions. Positive reactions are shown by macroscopic aggregation of the particles. Although the kit is intended for use primarily as a qualitative test, antibody levels may be titrated by doubling dilution. Agglutination patterns are interpreted by eye. Contents tests 500 tests R1 RPR antigen 1 x 2mL 5 x 2 ml R2 Positive control 1 x 1mL 2 x 1 ml R3 Negative control 1 x 1mL 2 x 1mL Dispensing bottle 1 2 Dispensing needle 1 2 Test cards (10 circles) Directions for use 2

3 Warnings and precautions For in - vitro diagnostic use only. All reagents contain sodium azide (less than 0.1% w/v). Waste fluids arising from use of the kit must be flushed with large quantities of water to avoid accumulation of potentially explosive compounds in laboratory plumbing. The control materials supplied are derived from human serum. They have been tested at donor level and found negative for Hepatitis B and C, and for HIV 1 and 2. However, they should be treated as if capable of transmitting disease. Specimens of human serum and plasma should be treated as microbiologically hazardous, and handled in accordance with the applicable regulations. Do not use the kit after its expiry date. Do not combine or interchange reagents from kits with different lot numbers. Storage. Store at 2 8 degrees C when not in use. Store bottles upright. Do not freeze. Shelf life is valid until date stated on kit label. Equipment required Properly calibrated and maintained pipette (or disposable droppers) to deliver a volume of 50 microlitres. Rotator to rotate test cards at 100 rpm, in a circle of diameter approximately 1 cm. Specimens Serum or plasma specimens should be free of blood cells and of obvious microbial contamination, gross haemolysis, turbidity, and lipaemia. They may be stored at 2-8 degrees C for up to 7 days before testing. Specimens needing longer storage should be frozen at 20 degrees C or lower. Frozen specimens should be thawed and well mixed before testing. Test method. Bring all reagents, controls, and specimens to room temperature prior to use. Do not use specimens that are grossly contaminated, excessively haemolysed, extremely turbid or lipaemic. Qualitative Test. 1. The kit positive and negative controls must be run with each run of tests. 2. Place 50µl of specimen or control into a circle on the test card. 3. Spread the specimen evenly over the test circle area. 4. Shake the vial of RPR antigen to ensure thorough mixing. 5. Attach the dropping needle to the plastic dropping bottle and take up the RPR antigen by suction. 6. Invert the dropping bottle and gently squeeze to expel air from the needle. 7. Holding the dropping bottle vertically over the test specimen, dispense a single drop of antigen. 8. Place test card on a card rotator and rotate at 100 rpm for 8 minutes. 9. Read and interpret results visually in good light. (See Interpretation. ) 10. Return unused antigen from dropper bottle to glass vial. 11. Clean out dropper bottle and needle with distilled water and allow to dry before re-using. 3

4 Semi-Quantitative Test. 1. Make doubling dilutions from undiluted to 1:16 in normal saline. 2. Place 50µl of each dilution in to a separate circle on the test card. 3. Spread each dilution evenly over the test circle. 4. Continue as from Qualitative Test section 3. The titre of the specimen is expressed as the reciprocal of the highest dilution showing aggregation of the carbon particles. Interpretation and assay validation Strong Reactive (SR): Large clumps of carbon particles with a clear background. Reactive (R) : Weak Reactive (WR): Large clumps of carbon particles somewhat more dispersed than in Strong Reactive. Small clumps of carbon particles with light grey background. Trace Reactive (TR): Slight clumping of carbon particles typically seen as a button of aggregates in the centre of the test circle or dispersed around the edge of the test circle. Non-Reactive (NR): Typically a smooth grey pattern or a button of non-aggregated carbon particles in the centre of the test circle. Reactive samples should be recorded as antibody positive and must (in view of the non-specific nature of the antibodies detected) be subjected to further tests to determine the presence or absence of specific antitreponemal antibody. For the assay to be valid the positive control provided should give a strong positive pattern and the negative control provided should give a clearly negative result. Performance characteristics. The following materials were independently tested to compare the performance of RPR 100 & 500 (3 different lots) with that of a reference reagent from CDC Atlanta, and a reagent from another manufacturer. Reference plasma panel (n=20) from CDC Atlanta Positive Test panel from a UK hospital (n=50) Normal donor plasma (n=50) WHO control All specimens gave 100% concordant qualitative results with all reagents. (95% confidence %) Titres of positive samples did not vary between reagents by more than one doubling dilution Precision and Accuracy For N = 10 assays of a positive sample %CV = 0% Accuracy = +/- 0% 4

5 Bibliography 1. Larsen SA.,Pettit, et coll., EDTA treated plasma in the rapid plasm regain card test and the toluidine red unheated serum test for serodiagnosis syphilis. J Clin Microbiology 1983;17; Portnoy J. Modifications of the rapid plasma regain (RPR) card test for syphilis, for use in large-scale testing.am J Clin Pathol;1963;40; Larsen S.A., Pope V., et coll., A manual of Tests for Syphilis 9 th Edition ;1998;

6 UK Use By DE Verwendbar bis ES Fecha de caducidad IT Utilizzare entro FR Utiliser jusque S Utgångsdatum DK Holdbar til P Fecha de caducidad UK Manufacturer DE Hersteller ES Fabricante IT Fabbricante FR Fabricant S Tillverkat av DK Producent P Fabricante UK In Vitro Diagnostic Medical Device DE In Vitro Diagnostikum ES Producto sanitario para diagnóstico in vitro IT Dispositivo medico-diagnostico in vitro FR Dispositif médical de diagnostic in vitro S In vitro-diagnostik DK Medicinsk udstyr til in vitro-diagnostik P Producto sanitario para diagnóstico in vitro UK Temperature limitation DE Zulässiger Temperaturbereich ES Limite de temperatura IT Limiti di temperatura FR Limites de température S Förvaring i temperaturområde DK Temperaturbegrænsning P Limite de temperatura UK Consult Instructions for Use- DE Gebrauchsanweisung beachten ES Consulte las instrucciones de uso IT Consultare le istruzioni per l'uso FR Consulter les instructions d'utilisation S Se bruksanvisningen DK Se brugsanvisning P Consulte las instrucciones de uso UK Batch code DE Chargenbezeichnung ES Codigo de lote IT Codice del lotto FR Code du lot S Lot nummer DK Lotnummer P Codigo de lote CE MOD1 2003/10 33