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

Bloomberg School of Public Health

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

Keywords

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

ASJC Scopus subject areas

Microbiology
Immunology
Infectious Diseases

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

In: Microbes and Infection, Vol. 11, No. 3, 03.2009, p. 419-423.

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

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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",

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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

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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.

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