Malaria Pf/pan antigen Rapid Test

Transcription

1 Malaria Pf/pan antigen Rapid Test Cat. No.:IVDTS003 Pkg.Size:10T/50T Intended use The Malaria Pf/pan antigen Rapid Test is a self-performing, qualitative, sandwich immunoassay, utilizing whole blood for the detection of P.falciparum specific histidine rich protein-2 (P.f HRP-2) and Pan specific pldh. The test may be used for differentiation of P.falciparum and other malarial species in human blood samples and for the follow up of anti-malarial therapy as well as for malaria endemic areas screening. For in vitro diagnostic use only. General Description Malaria remains one of the most serious tropical and subtropical diseases in many countries of the world. It is rampant in most areas of the tropics. Malaria is caused by a parasite that is transmitted from one person to another by the bite of infected Anopheles mosquitoes. There are four kinds of malaria that can infect humans: Plasmodium falciparum, P.vivax, P.ovale and P.malariae. Malaria also has been reported from blood transfusions or congenitally from mother to child. It is estimated to affect more than 500 million people causing between one and three million deaths every year. Principle Of The Test The Malaria Pf/pan antigen Rapid Test utilizes the principle of immunochoromatograhpy. As the test sample flows through the membrane assembly of the device after addition of the buffer, the marked monoclonal P.f specific HRP-2, Pan specific pldh colloidal gold conjugate antibodies combine the proteins in the lysed sample. This complex moves further on the membrane to the test region where it is immobilized by the P.f specific HRP-2 antibody/pan specific pldh which will lead to a formation of purple color line/s. While both the lines will appear at the test region in falciparum positive samples, only one line would appear in non-falciparum malaria positive sample. Absence of this color line/s in the test region indicates negative test result. The unreacted conjugate along with the rabbit globulin colloidal gold conjugate and unbound complex if any, move further on the membrane and are subsequently immobilized by anti-rabbit antibodies coated on the membrane at the control region, forming a pink/purple line. This control line serves to validate the test performance. Reagents And Materials Provided 1. Individual pouches, each containing: - Test kit (Strip/Cassette) - Desiccant pouch - Sample dropper (5µl) 2. Sample dilute (one bottle for each 25 tests, or one bottle for 10 test package) 3. Instructions for use Materials Required But Not Supplied 1. Alcohol pads 2. Lancets 3. Tube 1

2 Storage 1. Stored at 4 to 30 in the sealed foil pouch up to the expiration date. 2. Keep away from sunlight, moisture and heat. 3. DO NOT FREEZE. Specimen Collection And Preparation Collection by venipuncture 1. Collect the whole blood into the collection tube (using the suitable anti-coagulant) by venipuncture. 2. If the specimens are not immediately tested, they should be refrigerated at 2 C ~ 8 C. For storage periods greater than 24 hours, freezing is recommended. The frozen specimen more than 7 days is not recommended to use. The samples should be brought to room temperature prior to use. Collection using a lancet 1. Clean the area to be lanced with an alcohol swab. 2. Squeeze the end of the fingertip (or earlobe) and pierce with a sterile lancet. Wipe away the first drop of blood with sterile cotton. 3. Use a sample dropper, while gently squeezing the tube, immerse the open end in the blood drop and then gently release the pressure to draw blood into the dropper. Assay Procedure Allow the test kit, dilute and specimen to equilibrate to room temperature (10 C ~30 C) prior to testing. 1. Remove the test kit (Strip/Cassette) from the foil pouch by tearing at the notch and place it on a level surface. 2. Slowly vertically add 5 µl of whole blood to the sample well and then vertically add 4 drops (about 80ul) of dilute to the buffer well. 3. As the test begins to work, you will see purple color move across the strip/cassette in the center of the strip/cassette. 4. The best results are read in 15 minutes. The results are invalid after 15 minutes. Figure 1. Caution: 1. When adding lysed buffer, the bottle can not be level and touch the well. 2. The above interpreting time is based on reading the test results at room temperature of 10 ~30. If your room temperature is significantly lower than 10, then the interpreting time should be properly increased. 3. The test kit should be used in 60 minutes after open the pouch, in void of losing efficacy. Quality Control Though there is an internal procedural control line in the test device of control region, the use of external controls is strongly recommended as good laboratory testing practice to confirm the test procedure and to verify proper test performance. Positive and negative control should give the expected results. When testing the positive and negative control, the same assay procedure should be adopted. 2

3 Validation of The Test Test one specimen by 10 kits from one batch in the same time, operated strictly under the user guide, by one operator in the same lab, the color to be consistent, no obvious different. Interpretation of Results Positive: 1) The presence of two color line ( T1 and C ) indicates a positive result for a infection with P. falciparum. 2) The presence of three color lines ( T1, T2 and C ) indicates a positive result for P. falciparum monoinfection or mixed infection among the four kinds of plasmodium. 3) The presence of two color lines ( T2 and C ) indicates a negative result for P. falciparum and a positive result for Non-P. falciparum (P. vivax, P. malariae or P.ovale) monoinfection or mixed infection. Figure 2. Negative: The presence of only one line ( C ) indicates a negative result. Figure 3. Invalid: If the color line ( C ) is not visible within the reading window after performing the test, the result is considered invalid. The directions may not have been followed correctly or the test may have deteriorated. It is recommended that the specimen be retested. Figure 4. 3

4 Notes: 1. The used whole blood sample may appear a red background in the reading window, if it does not cover the test line, the result is valid. 2. The pink line may appear color shades in test region (T), provided within the observation time, the result should be determined positive. 3. When the plasmodium at the higher level, the test line will be strong, but the control line is weak, this result should be considered acceptable. 4. If the sample with high level of plasmodium, it will be caused hook effect, the results might be false negative, in this situation, the sample should be diluted. Detection Range The Detection limit is plasmodium/μl (Whold Blood). Sensitivity and Specificity The samples term: 30 falciparum malaria whole blood samples, 30 Non falciparum malaria whold blood sample, 100 health whole blood sample. The level of the plasmodium had been confirmed by staining microscopic examination method. The results as below: Pf positive coincidence rate is 100%, Pan positive coincidence rate is 100%, the Negative coincidence rate is 97%. Table 1. Precautions 1. This kit is for in vitro use only. Do not swallow. 2. Discard after first use. The kit cannot be used more than once. 3. Do not use the kit beyond the expiration date. 4. The device and the buffer of different lots must not be mixed and used. 5. Do not use the kit if the pouch is punctured or not well sealed. 6. Decontaminate and dispose of all specimens, reaction kits and potentially contaminated materials, as if they were infectious waste, in a biohazard container. The used-kit has the infectious risk. The process of disposing the diagnostics must follow the local infectious disposal law or laboratory regulation 7. Wear protective gloves while handling specimens. Wash hands thoroughly afterwards. Avoid splashing or aerosol formation. Clean up the spills 4

5 8. Keep out of the reach of children. Limitations The test is limited to the qualitative detection of P. falciparum specific histidine rich protein-2 (P.f HRP-2) and Pan specific pldh. Although the test is accurate in detecting antigens of Malaria P.f/Pan, a low incidence of false results can occur. Other clinically available test are required if questionable results are obtained. As with all diagnostic tests, a definitive clinical diagnosis should not be based on the results of a single test, but should only be made by the physician after all clinical and laboratory findings have been evaluated. REFERENCES 1. The Diagnostic Criteria of Malaria, Ministry of Health of The People's Republic of China, the Healthy Industry Standard of the People's Republic of China, WS The Research Progress of Detection Technique to Malaria,. Yu Dan, South China J Prev Med, December 2006, Vol 32,No The Research Progress of Diagnostic Kit for Malaria, Li Xueping, Lab Med Clin, January 2009,Vol.6,No.1. 5