Volume 28 Issue 1
Jan.  2015
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GUO Qian, YU Yan, ZHU Yan Ling, ZHAO Xiu Qin, LIU Zhi Guang, ZHANG Yuan Yuan, LI Gui Lian, WEI Jian Hao, WU Yi Mou, WAN Kang Lin. Rapid Detection of Rifampin-resistant Clinical Isolates of Mycobacterium tuberculosis by Reverse Dot Blot Hybridization[J]. Biomedical and Environmental Sciences, 2015, 28(1): 25-35. doi: 10.3967/bes2015.003
Citation: GUO Qian, YU Yan, ZHU Yan Ling, ZHAO Xiu Qin, LIU Zhi Guang, ZHANG Yuan Yuan, LI Gui Lian, WEI Jian Hao, WU Yi Mou, WAN Kang Lin. Rapid Detection of Rifampin-resistant Clinical Isolates of Mycobacterium tuberculosis by Reverse Dot Blot Hybridization[J]. Biomedical and Environmental Sciences, 2015, 28(1): 25-35. doi: 10.3967/bes2015.003

Rapid Detection of Rifampin-resistant Clinical Isolates of Mycobacterium tuberculosis by Reverse Dot Blot Hybridization

doi: 10.3967/bes2015.003
Funds:  This work was funded by the projects 2013ZX10003002-001 and 2013ZX10003006-002-001 of Chinese National Key Program of Mega Infectious Disease of the National 12th Five-Year Plan
  • Objective A PCR-reverse dot blot hybridization (RDBH) assay was developed for rapid detection of rpoB gene mutations in‘hot mutation region’ of Mycobacterium tuberculosis (M. tuberculosis).
    Methods 12 oligonucleotide probes based on the wild-type and mutant genotype rpoB sequences of M. tuberculosis were designed to screen the most frequent wild-type and mutant genotypes for diagnosing RIF resistance. 300 M. tuberculosis clinical isolates were detected by RDBH, conventional drug-susceptibility testing (DST) and DNA sequencing to evaluate the RDBH assay.
    Results The sensitivity and specificity of the RDBH assay were 91.2%(165/181) and 98.3%(117/119), respectively, as compared to DST. When compared with DNA sequencing, the accuracy, positive predictive value (PPV) and negative predictive value (NPV) of the RDBH assay were 97.7%(293/300), 98.2%(164/167), and 97.0%(129/133), respectively. Furthermore, the results indicated that the most common mutations were in codons 531 (48.6%), 526 (25.4%), 516 (8.8%), and 511 (6.6%), and the combinative mutation rate was 15 (8.3%). One and two strains of insertion and deletion were found among all strains, respectively.
    Conclusion Our findings demonstrate that the RDBH assay is a rapid, simple and sensitive method for diagnosing RIF-resistant tuberculosis.
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通讯作者: 陈斌, bchen63@163.com
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    沈阳化工大学材料科学与工程学院 沈阳 110142

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Rapid Detection of Rifampin-resistant Clinical Isolates of Mycobacterium tuberculosis by Reverse Dot Blot Hybridization

doi: 10.3967/bes2015.003
Funds:  This work was funded by the projects 2013ZX10003002-001 and 2013ZX10003006-002-001 of Chinese National Key Program of Mega Infectious Disease of the National 12th Five-Year Plan

Abstract: Objective A PCR-reverse dot blot hybridization (RDBH) assay was developed for rapid detection of rpoB gene mutations in‘hot mutation region’ of Mycobacterium tuberculosis (M. tuberculosis).
Methods 12 oligonucleotide probes based on the wild-type and mutant genotype rpoB sequences of M. tuberculosis were designed to screen the most frequent wild-type and mutant genotypes for diagnosing RIF resistance. 300 M. tuberculosis clinical isolates were detected by RDBH, conventional drug-susceptibility testing (DST) and DNA sequencing to evaluate the RDBH assay.
Results The sensitivity and specificity of the RDBH assay were 91.2%(165/181) and 98.3%(117/119), respectively, as compared to DST. When compared with DNA sequencing, the accuracy, positive predictive value (PPV) and negative predictive value (NPV) of the RDBH assay were 97.7%(293/300), 98.2%(164/167), and 97.0%(129/133), respectively. Furthermore, the results indicated that the most common mutations were in codons 531 (48.6%), 526 (25.4%), 516 (8.8%), and 511 (6.6%), and the combinative mutation rate was 15 (8.3%). One and two strains of insertion and deletion were found among all strains, respectively.
Conclusion Our findings demonstrate that the RDBH assay is a rapid, simple and sensitive method for diagnosing RIF-resistant tuberculosis.

GUO Qian, YU Yan, ZHU Yan Ling, ZHAO Xiu Qin, LIU Zhi Guang, ZHANG Yuan Yuan, LI Gui Lian, WEI Jian Hao, WU Yi Mou, WAN Kang Lin. Rapid Detection of Rifampin-resistant Clinical Isolates of Mycobacterium tuberculosis by Reverse Dot Blot Hybridization[J]. Biomedical and Environmental Sciences, 2015, 28(1): 25-35. doi: 10.3967/bes2015.003
Citation: GUO Qian, YU Yan, ZHU Yan Ling, ZHAO Xiu Qin, LIU Zhi Guang, ZHANG Yuan Yuan, LI Gui Lian, WEI Jian Hao, WU Yi Mou, WAN Kang Lin. Rapid Detection of Rifampin-resistant Clinical Isolates of Mycobacterium tuberculosis by Reverse Dot Blot Hybridization[J]. Biomedical and Environmental Sciences, 2015, 28(1): 25-35. doi: 10.3967/bes2015.003

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