Detection of Mycobacterium tuberculosis complex in formalin-fixed, paraffin-embedded tissue by Multiplex Polymerase Chain Reaction Assay
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Abstract
Tuberculosis (TB) is a common infectious disease and a serious public health problem in Thailand. The causative agents of TB are a group of closely related bacteria known as the Mycobacterium tuberculosis complex (MTBC). One accepted method for the diagnosis of tuberculous infection is the detection of their DNA by conventional polymerase chain reaction (C-PCR) assay. However, this method is complicated and time-consuming, and thus unsuitable for mass screening. A simpler and faster multiplex PCR (M-PCR) assay has been developed to overcome these problems. The objective of this study is to prove that detection of MTBC DNA from formalin-fixed, paraffin-embedded (FFPE) tissue by M-PCR assay can be acceptable in comparison to the previous C-PCR assay. Paraffin-embedded tissue samples of one hundred and fifty suspected cases of tuberculosis referred to the Institute of Pathology, Department of Medical Services were retrieved for DNA extraction. IS6110 and B-globin gene were examined by M-PCR and C-PCR assays for the detection of M. tuberculosis complex. The results of the M-PCR assay agreed with the C-PCR assay (K=1.00, 95% CI 1.00-1.00). In conclusion, M-PCR assay is a simpler, faster, and less costly method that can be an efficient and effective alternative to C-PCR assay for the DNA detection of M. tuberculosis complex from formalinfixed, paraffin-embedded (FFPE) tissue.
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