Authors : Imadeldin Aradaib, Mohamed B. Elamin and Abdelatif Ashmaig
Abstract: Rapid and accurate diagnosis of Mycobacterium tuberculosis complex is essential for the treatment and outcome of the disease as well as prevention of its further transmission. In the present study, a nested Polymerase Chain Reaction (nPCR)-based detection assay is described for rapid detection of tuberculosis. The IS6110 sequence of the M. tuberculosis complex was used as a target gene for PCR in two amplification steps. The first round of amplification produced a 567 bp primary PCR product. The second round of amplification, using internal (nested) primers, resulted in a 310 bp nested PCR product. The nPCR provides rapid, sensitive and specific detection of the organism in clinical samples and a variety of different body fluids and aspirates. Clinical samples were collected from infected patients admitted to the National Ribat University Medical Teaching Hospital, Khartoum, Sudan. The sensitivity of the assay indicated that the nPCR detected as little as a single copy of the target DNA. However, application of this nPCR to DNA from other related bacteria including M. flavescens, M. duvalii and M. paratuberculosis did not amplify the primary or the nested PCR products. The described nPCR could be used as a valuable tool for early differential diagnosis of tuberculosis in biological fluids from suspected patients. In addition, the assay should be recommended for inclusion during epidemiological surveys, treatment and control programs of the disease.
Imadeldin Aradaib, Mohamed B. Elamin and Abdelatif Ashmaig, 2009. Application of Polymerase Chain Reaction for Rapid Detection of Mycobacterium tuberculosis Complex in Biological Fluids. International Journal of Tropical Medicine, 4: 123-128.