[Establishment of the standard operation program on multiple loci variable numbers tandem repeats analysis typing on Mycobacterium tuberculosis].
OBJECTIVE
To establish and evaluate the standardized protocol of multiple loci variable numbers tandem repeats (VNTR) analysis (MLVA) for genotyping Mycobacterium tuberculosis (M. tuberculosis).


METHODS
15 VNTR loci were chosen for genotyping 54 Chinese M. tuberculosis strains by PCR-electrophoresis-based VNTR analysis and the results were analyzed by software BioNumerics (Version 5.0).


RESULTS
MLVA method was successfully established and standardized, including the standard protocol for bacterial culture, DNA isolation,PCR and agarose gel electrophoresis and the software analysis. 15 VNTR loci were confirmed, including ETRA, ETRB, ETRC, ETRD, ETRE, MIRU10, MIRU16, MIRU23, MIRU26, MIRU27, MIRU39, MIRU40, Mtub21, Mtub30 and Mtub39, to be suitable for MLVA analysis of M. tuberculosis.


CONCLUSIONS
The standardized MLVA method has been established successfully. This method is simple and has powerful capacity for genotyping and strain differentiation, can be used for the network surveillance on pathogens of M. tuberculosis, and the data are comparable between laboratories. It is valuable for tracing the source and studying the trend of prevalence during investigation of M. tuberculosis infections.
Lu
Bing
B
State Key Laboratory for Infectious Diseases Prevention and Control, National Institute for Communicable Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing 102206, China.
Pourcel
Christine
C
Liu
Jing-Hua
JH
Dong
Hai-Yan
HY
Zhang
Yuan-Yuan
YY
Jiang
Yi
Y
Liu
Zhi-Guang
ZG
Zhao
Xiu-Qin
XQ
Wan
Kang-Lin
KL
D015373:Bacterial Typing Techniques / Q000379:methods*; D005838:Genotype; D018598:Minisatellite Repeats; D009169:Mycobacterium tuberculosis / Q000145:classification / Q000235:genetics* / Q000302:isolation & purification*; D012150:Polymorphism, Restriction Fragment Length