A rapid and simple method for identifying Mycobacterium tuberculosis W-Beijing strains based on detection of a unique mutation in Rv0927c by PCR-SSCP

Xin Jiang; Chanyi Lu; Feng Gao; Feifei Wang; Wenhong Zhang; Isabel Portugal; Peng Xu; Honghai Wang; Ying Zhang

doi:10.1016/j.micinf.2009.01.008

A rapid and simple method for identifying Mycobacterium tuberculosis W-Beijing strains based on detection of a unique mutation in Rv0927c by PCR-SSCP

Xin Jiang, Chanyi Lu, Feng Gao, Feifei Wang, Wenhong Zhang, Isabel Portugal, Peng Xu, Honghai Wang, Ying Zhang

Research output: Contribution to journal › Article › peer-review

Abstract

Recently we have found that W-Beijing Mycobacterium tuberculosis strains have a unique in-frame trinucleotide (AGC) deletion at position 421 of Rv0927c and a -127G → A mutation in Rv0927c-pstS3 intergenic region. Based on detecting the 421 trinucleotide deletion of these two mutations which can alter the ssDNA conformation more extensively than the other, we developed a PCR-SSCP method for rapid identification of W-Beijing strains among non-Beijing strains. Altogether, 104 clinical isolates were analyzed, including 68 W-Beijing strains and 36 non-Beijing strains. We found that PCR-SSCP successfully differentiated all the W-Beijing strains from the non-Beijing strains. In addition, we unexpectedly discovered that SDS-PAGE protein gels had better resolving power than conventional TBE polyacrylamide gel in detecting the AGC deletion mutation in the SSCP analysis.

Original language	English (US)
Pages (from-to)	419-423
Number of pages	5
Journal	Microbes and Infection
Volume	11
Issue number	3
DOIs	https://doi.org/10.1016/j.micinf.2009.01.008
State	Published - Mar 2009
Externally published	Yes

Keywords

Mycobacterium tuberculosis
PCR-SSCP
Rv0927c
Strain identification
W-Beijing strain

ASJC Scopus subject areas

Microbiology
Immunology
Infectious Diseases

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10.1016/j.micinf.2009.01.008

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title = "A rapid and simple method for identifying Mycobacterium tuberculosis W-Beijing strains based on detection of a unique mutation in Rv0927c by PCR-SSCP",

abstract = "Recently we have found that W-Beijing Mycobacterium tuberculosis strains have a unique in-frame trinucleotide (AGC) deletion at position 421 of Rv0927c and a -127G → A mutation in Rv0927c-pstS3 intergenic region. Based on detecting the 421 trinucleotide deletion of these two mutations which can alter the ssDNA conformation more extensively than the other, we developed a PCR-SSCP method for rapid identification of W-Beijing strains among non-Beijing strains. Altogether, 104 clinical isolates were analyzed, including 68 W-Beijing strains and 36 non-Beijing strains. We found that PCR-SSCP successfully differentiated all the W-Beijing strains from the non-Beijing strains. In addition, we unexpectedly discovered that SDS-PAGE protein gels had better resolving power than conventional TBE polyacrylamide gel in detecting the AGC deletion mutation in the SSCP analysis.",

keywords = "Mycobacterium tuberculosis, PCR-SSCP, Rv0927c, Strain identification, W-Beijing strain",

author = "Xin Jiang and Chanyi Lu and Feng Gao and Feifei Wang and Wenhong Zhang and Isabel Portugal and Peng Xu and Honghai Wang and Ying Zhang",

note = "Funding Information: Supported by Leading Academic Discipline Project of Shanghai Municipal Education Commission, Project Number:J50301 (Xin Jiang, Shanghai University of T.C.M) and Major State Basic Research (973) Program (No. 2005CB523102). YZ was supported by NIH grant AI44063. ",

year = "2009",

month = mar,

doi = "10.1016/j.micinf.2009.01.008",

language = "English (US)",

volume = "11",

pages = "419--423",

journal = "Microbes and Infection",

issn = "1286-4579",

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TY - JOUR

T1 - A rapid and simple method for identifying Mycobacterium tuberculosis W-Beijing strains based on detection of a unique mutation in Rv0927c by PCR-SSCP

AU - Jiang, Xin

AU - Lu, Chanyi

AU - Gao, Feng

AU - Wang, Feifei

AU - Zhang, Wenhong

AU - Portugal, Isabel

AU - Xu, Peng

AU - Wang, Honghai

AU - Zhang, Ying

N1 - Funding Information: Supported by Leading Academic Discipline Project of Shanghai Municipal Education Commission, Project Number:J50301 (Xin Jiang, Shanghai University of T.C.M) and Major State Basic Research (973) Program (No. 2005CB523102). YZ was supported by NIH grant AI44063.

PY - 2009/3

Y1 - 2009/3

N2 - Recently we have found that W-Beijing Mycobacterium tuberculosis strains have a unique in-frame trinucleotide (AGC) deletion at position 421 of Rv0927c and a -127G → A mutation in Rv0927c-pstS3 intergenic region. Based on detecting the 421 trinucleotide deletion of these two mutations which can alter the ssDNA conformation more extensively than the other, we developed a PCR-SSCP method for rapid identification of W-Beijing strains among non-Beijing strains. Altogether, 104 clinical isolates were analyzed, including 68 W-Beijing strains and 36 non-Beijing strains. We found that PCR-SSCP successfully differentiated all the W-Beijing strains from the non-Beijing strains. In addition, we unexpectedly discovered that SDS-PAGE protein gels had better resolving power than conventional TBE polyacrylamide gel in detecting the AGC deletion mutation in the SSCP analysis.

AB - Recently we have found that W-Beijing Mycobacterium tuberculosis strains have a unique in-frame trinucleotide (AGC) deletion at position 421 of Rv0927c and a -127G → A mutation in Rv0927c-pstS3 intergenic region. Based on detecting the 421 trinucleotide deletion of these two mutations which can alter the ssDNA conformation more extensively than the other, we developed a PCR-SSCP method for rapid identification of W-Beijing strains among non-Beijing strains. Altogether, 104 clinical isolates were analyzed, including 68 W-Beijing strains and 36 non-Beijing strains. We found that PCR-SSCP successfully differentiated all the W-Beijing strains from the non-Beijing strains. In addition, we unexpectedly discovered that SDS-PAGE protein gels had better resolving power than conventional TBE polyacrylamide gel in detecting the AGC deletion mutation in the SSCP analysis.

KW - Mycobacterium tuberculosis

KW - PCR-SSCP

KW - Rv0927c

KW - Strain identification

KW - W-Beijing strain

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A rapid and simple method for identifying Mycobacterium tuberculosis W-Beijing strains based on detection of a unique mutation in Rv0927c by PCR-SSCP

Abstract

Keywords

ASJC Scopus subject areas

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