Evaluation of PCR-ELISA method for Detection of Mycobacterium tuberculosis in Clinical Specimens

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Objective: Tuberculosis (TB) is one of the major causes of death worldwide. In the controlling of Tuberculosis (TB), early diagnosis of disease caused by Mycobacterium tuberculosis complex (MTBC) is vital. Hence, nucleic acid amplification tests (NAAT) for rapid diagnosis of TB came to be used widely in recent years. PCR-ELISA is a method which based on nucleic acid amplification based methods that are used for the direct detection of MTBC in clinical samples. The aim of study, was to evaluate the performance of ProDect MYCOBACTERIUM TUBERCULOSIS (bcs Biotect S.p.A., Cagliari, Italia), a commercial test based on PCR-ELISA for the detection of MTBC in pulmonary and extrapulmonary clinical samples. Material and Methods: 100 clinical samples (76 pulmonary and 24 extrapulmonary) from patients suspected of TB were tested by microscopy, culture and ProDect MYCOBACTERIUM TUBERCULOSIS PCR-ELISA methods. In this test, detection of the amplified product is performed on microplates using DNA enzyme immunoassay. The DNA enzyme immunoassay is based on the hybridization of amplified DNA with a single-stranded DNA probe, coated on the wall of the microtitre plate wells by a streptavidin-biotin bond. The hybrid between probe and DNA being examined is detected by using an anti-DNA mouse monoclonal antibody. Results: When PCR-ELISA test results were evaluated according to culture results which is taken as the gold standard, the sensitivity and, specificity were 68.2% and 94.1% , respectively, for whole specimens. The sensitivity, specificity were 77.1% , 93.5% for pulmonary specimens and 61.9% , 100% for extrapulmonary specimens, respectively. For smear positive pulmonary specimens, the sensitivity and specificity were found to be 83.3% and 100% , respectively. Conclusion: PCR-ELISA method can be utilized cost-effectively, providing results with the use of less equipment, as a routine nucleic amplification test for the rapid diagnosis of tuberculosis in a mycobacteriology laboratory, specifically for smear positive pulmonary samples.

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Mycobacterium tuberculosis , PCR , ELISA , tuberculosis , diagnosis

Kocatepe Medical Journal

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Kocatepe Medical Journal