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Evaluation of efficiency of nested multiplex allele‐specific PCR assay for detection of multidrug resistant tuberculosis directly from sputum samples
Author(s) -
Mistri S.K.,
Sultana M.,
Kamal S.M.M.,
Alam M.M.,
Irin F.,
Nessa J.,
Ahsan C.R.,
Yasmin M.
Publication year - 2016
Publication title -
letters in applied microbiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.698
H-Index - 110
eISSN - 1472-765X
pISSN - 0266-8254
DOI - 10.1111/lam.12564
Subject(s) - sputum , nested polymerase chain reaction , tuberculosis , rpob , mycobacterium tuberculosis , multiplex , polymerase chain reaction , biology , multiplex polymerase chain reaction , virology , drug resistance , microbiology and biotechnology , medicine , gene , genetics , pathology
Abstract For an effective control of tuberculosis, rapid detection of multidrug resistant tuberculosis ( MDR ‐ TB ) is necessary. Therefore, we developed a modified nested multiplex allele‐specific polymerase chain reaction ( MAS ‐ PCR ) method that enables rapid MDR ‐ TB detection directly from sputum samples. The efficacy of this method was evaluated using 79 sputum samples collected from suspected tuberculosis patients. The performance of nested MAS ‐ PCR method was compared with other MDR ‐ TB detection methods like drug susceptibility testing ( DST ) and DNA sequencing. As rifampicin ( RIF ) resistance conforms to MDR ‐ TB in greater than 90% cases, only the presence of RIF ‐associated mutations in rpoB gene was determined by DNA sequencing and nested MAS ‐ PCR to detect MDR ‐ TB . The concordance between nested MAS ‐ PCR and DNA sequencing results was found to be 96·3%. When compared with DST , the sensitivity and specificity of nested MAS ‐ PCR for RIF ‐resistance detection were determined to be 92·9 and 100% respectively. Significance and Impact of the Study For developing‐ and high‐ TB burden countries, molecular‐based tests have been recommended by the World Health Organization for rapid detection of MDR ‐ TB . The results of this study indicate that, nested MAS ‐ PCR assay might be a practical and relatively cost effective molecular method for rapid detection of MDR ‐ TB from suspected sputum samples in developing countries with resource poor settings.

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