Amplification of Mycobacterium tuberculosis in clinical samples and detection using the EIA format

Abstract

A test based on the polymerase chain reaction was used for the detection of Mycobacterium tuberculosis in clinical samples. A 400 bp sequence of the mtp40 that was specific for M. tuberculosis was amplified and detected using the enzyme-linked immunosorbent assay (ElA) detection format. We tested 61 clinical samples of various types from 44 patients with suspected tuberculosis. These samples included cerebrospinal fluids, pleural tap/fluids, sputum, nasopharyngeal aspirates, gastric lavage, peritoneal fluids, urine, tracheal aspirates, serum and blood. Only sputum samples were first decontaminated. All samples were heated to lyse the cells and the extracted DNA was cleaned with phenol-chloroform. The specificity of the probe was tested with amplified products of primers for IS986. In this study, 41 samples were positive with the EIA format whereas 32 samples were positive based on analysis by gel electrophoresis.