| Title | Rapid diagnosis of isoniazid resistant mycobacterium tuberculosis by high resolution melting (HRM) assay |
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| Authors | Chan, Ming-yan
陳明恩 |
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| Issue Date | 2012 |
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| Publisher | The University of Hong Kong (Pokfulam, Hong Kong) |
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| Abstract | Mycobacterium tuberculosis (MTB) is a major infective agent causing human tuberculosis (TB) in the worldwide. Although tuberculosis can be treated by a six-month course of antibiotics, the prevalence of extensively drug-resistance TB (XDR-TB) made the disease becomes a global health problem. In addition to the conventional MTB detection methods, molecular methods become significant in drug resistant MTB detection which can enhance effective drug treatment.
In this study, 200 MTB respiratory specimens were collected from patients with suspected tuberculosis in Tuen Mun Hospital in Hong Kong. Based on the culture method as a gold standard for MTB detection, the presence of MTB in clinical samples was determined by IS6110single tube nested real-time PCR. In addition, by using High Resolution Melting (HRM) analysis, the presence of mutant type KatG315 gene for detecting isoniazid resistant MTB was determined.
Among 66 MTB culture positive samples, 10 samples had positive acid fast bacilli (AFB) smears giving the diagnostic sensitivity 15.1%. IS6110 single tube nested PCR was amplified in 51 specimens giving 77.2% MTB detection sensitivity and 97.8% specificity. Among 51 samples positive for IS6110 PCR, 66.7% showed successful amplification in subsequent KatG-HRM assay. Two samples were confirmed to be isoniazid (INH) resistance in Public Health Laboratory Centre (PHLC). However, there was only one sample showing detectable KatG315 mutation in clinical specimen by using HRM while the other was only detected in the corresponding culture isolate.
From the result of this study, single tube nested real-time PCR demonstrated MTB detection in clinical samples and INH resistant strain with KatG315mutationcan be detected by HRM analysis. Early detection of mycobacteria allow earlier treatment of the patient, thus transmission of the disease can be controlled. |
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| Degree | Master of Medical Sciences |
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| Subject | Mycobacterium tuberculosis - Diagnosis. |
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| Dept/Program | Microbiology |
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| DC Field | Value |
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| dc.contributor.author | Chan, Ming-yan |
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| dc.contributor.author | 陳明恩 |
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| dc.date.hkucongregation | 2012 |
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| dc.date.issued | 2012 |
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| dc.description.abstract | Mycobacterium tuberculosis (MTB) is a major infective agent causing human tuberculosis (TB) in the worldwide. Although tuberculosis can be treated by a six-month course of antibiotics, the prevalence of extensively drug-resistance TB (XDR-TB) made the disease becomes a global health problem. In addition to the conventional MTB detection methods, molecular methods become significant in drug resistant MTB detection which can enhance effective drug treatment.
In this study, 200 MTB respiratory specimens were collected from patients with suspected tuberculosis in Tuen Mun Hospital in Hong Kong. Based on the culture method as a gold standard for MTB detection, the presence of MTB in clinical samples was determined by IS6110single tube nested real-time PCR. In addition, by using High Resolution Melting (HRM) analysis, the presence of mutant type KatG315 gene for detecting isoniazid resistant MTB was determined.
Among 66 MTB culture positive samples, 10 samples had positive acid fast bacilli (AFB) smears giving the diagnostic sensitivity 15.1%. IS6110 single tube nested PCR was amplified in 51 specimens giving 77.2% MTB detection sensitivity and 97.8% specificity. Among 51 samples positive for IS6110 PCR, 66.7% showed successful amplification in subsequent KatG-HRM assay. Two samples were confirmed to be isoniazid (INH) resistance in Public Health Laboratory Centre (PHLC). However, there was only one sample showing detectable KatG315 mutation in clinical specimen by using HRM while the other was only detected in the corresponding culture isolate.
From the result of this study, single tube nested real-time PCR demonstrated MTB detection in clinical samples and INH resistant strain with KatG315mutationcan be detected by HRM analysis. Early detection of mycobacteria allow earlier treatment of the patient, thus transmission of the disease can be controlled. |
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| dc.description.nature | published_or_final_version |
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| dc.description.thesisdiscipline | Microbiology |
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| dc.description.thesislevel | master's |
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| dc.description.thesisname | Master of Medical Sciences |
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| dc.identifier.hkul | b4833340 |
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| dc.language | eng |
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| dc.publisher | The University of Hong Kong (Pokfulam, Hong Kong) |
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| dc.relation.ispartof | HKU Theses Online (HKUTO) |
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| dc.rights | The author retains all proprietary rights, (such as patent rights) and the right to use in future works. |
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| dc.rights | Creative Commons: Attribution 3.0 Hong Kong License |
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| dc.source.uri | http://hub.hku.hk/bib/B48333402 |
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| dc.subject.lcsh | Mycobacterium tuberculosis - Diagnosis. |
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| dc.title | Rapid diagnosis of isoniazid resistant mycobacterium tuberculosis by high resolution melting (HRM) assay |
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| dc.type | PG_Thesis |
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