Rapid identification of mycobacteria and rapid detection of drug resistance in mycobacterium tuberculosis in cultured isolates and in respiratory specimens

Wing Cheong Yam, Kit Hang Siu

Research output: Chapter in book / Conference proceedingChapter in an edited book (as author)Academic researchpeer-review

4 Citations (Scopus)

Abstract

Recent advances in molecular biology and better understanding of the genetic basis of drug resistance have allowed rapid identification of mycobacteria and rapid detection of drug resistance of Mycobacterium tuberculosis present in cultured isolates or in respiratory specimens. In this chapter, several simple nucleic acid amplification-based techniques are introduced as molecular approach for clinical diagnosis of tuberculosis. A one-tube nested IS6110-based polymerase chain reaction (PCR) is used for M. tuberculosis complex identification; the use of a multiplex allele-specific PCR is demonstrated to detect the isoniazid resistance; PCR-sequencing assays are applied for rifampicin and ofloxacin resistance detection and 16S rDNA sequencing is utilized for identification of mycobacterial species from cultures of acid fast bacilli (AFB). Despite the high specificity and sensitivity of the molecular techniques, mycobacterial culture remains the Gold Standard for tuberculosis diagnosis. Negative results of molecular tests never preclude the infection or the presence of drug resistance. These technological advancements are, therefore, not intended to replace the conventional tests, but rather have major complementary roles in tuberculosis diagnosis.
Original languageEnglish
Title of host publicationPCR Detection of Microbial Pathogens
Pages171-199
Number of pages29
DOIs
Publication statusPublished - 1 Jan 2013
Externally publishedYes

Publication series

NameMethods in Molecular Biology
Volume943
ISSN (Print)1064-3745

Keywords

  • 16S rDNA sequencing
  • COBAS TaqMan MTB Test ® ®
  • IS6110 PCR
  • Isoniazid resistance
  • Multiplex allele-specific PCR
  • Mycobacterium tuberculosis
  • Ofloxacin resistance
  • PCR sequencing
  • Rifampicin resistance

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

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