Polymerase Chain Reaction using the MPB64 fragment for detection of mycobacterium tuberculosis complex DNA in suspected TB cases

Abstract

Background: Various primers have been used for detection of Mycobacterium tuberculosis complex (MTBC) DNA using Polymerase chain reaction (PCR). The MPB64 gene has been demonstrated to be a highly specific target for MTBC detection. Aims & Objectives: To detect MTBC DNA using MPB64 primers in clinical specimens of Nepalese patients suspected to have tuberculosis (TB). Materials and Methods: DNA was extracted from 44 clinical specimens of patients suspected of having TB and MPB64 targeted PCR was performed. Results: Bands comprising 240 base pair (bp) of MPB64 sequence were present in 15 of the 44 clinical samples, of which 4 were pulmonary and the remainders were extra-pulmonary samples. The overall positivity of MPB64 targeted PCR was 34.1%. Conclusion: PCR targeting the MPB64 fragment has a potential of detecting the MTBC DNA and has potentially valuable clinical applications in early detection of TB in Nepal.

Modified genome comparison method: a new approach for identification of specific targets in molecular diagnostic tests using Mycobacterium tuberculosis complex as an example

“…Another diagnostic target sequence which many studies are based on, is mpb64 [ 28 , 37 , 38 ]. This sequence has 687 bp length [ 39 ].…”

Evaluation of multiplex PCR using MPB64 and IS6110 primers for rapid diagnosis of tuberculous meningitis

“…Similarly, MPB64 has been demonstrated to be highly specific for Mycobacterium tuberculosis complex [23] and other studies have reported sensitivity and specificity of 75e90% and 100% respectively [9,10,19,23]. The literature regarding the evaluation of MPB64 in patients of TBM in our region is scanty [9,24].…”

“…MPB64 primer targets and amplifies the gene MPB64, producing 240 bp product and IS6110 primer amplifies insertion sequence IS6110 of M. tuberculosis complex, which produces 123 bp product. Primers for both sequences were opted from previous studies [6,9,15] and the PCR conditions were optimized for multiplex PCR. The primers were analyzed using NCBI BLAST with insertion sequence IS6110 (Gene bank ID KP844721.1) and MPB64 gene (Gene bank ID 885925) before using them.…”