TY - CHAP
T1 - Rapid identification of mycobacteria and rapid detection of drug resistance in mycobacterium tuberculosis in cultured isolates and in respiratory specimens
AU - Yam, Wing Cheong
AU - Siu, Kit Hang
PY - 2013/1/1
Y1 - 2013/1/1
N2 - 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.
AB - 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.
KW - 16S rDNA sequencing
KW - COBAS TaqMan MTB Test ® ®
KW - IS6110 PCR
KW - Isoniazid resistance
KW - Multiplex allele-specific PCR
KW - Mycobacterium tuberculosis
KW - Ofloxacin resistance
KW - PCR sequencing
KW - Rifampicin resistance
UR - http://www.scopus.com/inward/record.url?scp=84870561676&partnerID=8YFLogxK
U2 - 10.1007/978-1-60327-353-4_12
DO - 10.1007/978-1-60327-353-4_12
M3 - Chapter in an edited book (as author)
C2 - 23104290
SN - 9781603273527
T3 - Methods in Molecular Biology
SP - 171
EP - 199
BT - PCR Detection of Microbial Pathogens
ER -