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Article: Rapid detection of rifampicin- and isoniazid-resistant mycobacterium tuberculosis by high-resolution melting analysis

TitleRapid detection of rifampicin- and isoniazid-resistant mycobacterium tuberculosis by high-resolution melting analysis
Authors
Issue Date2010
PublisherAmerican Society for Microbiology.
Citation
Journal Of Clinical Microbiology, 2010, v. 48 n. 4, p. 1047-1054 How to Cite?
AbstractWe have developed a high-resolution melting (HRM) assay to scan for mutations in the rpoB, inhA, ahpC, and katG genes and/or promoter regions for the detection of rifampin and isoniazid resistance in Mycobacterium tuberculosis. For assay development, 23 drug-resistant isolates of M. tuberculosis having 29 different mutations, together with 40 drug-susceptible isolates, were utilized. All 29 mutations were accurately detected by our assay. We further validated the assay with a series of 59 samples tested in a blind manner. All sequence alterations that were within the regions targeted by the HRM assay were correctly identified. Compared against results of DNA sequencing, the sensitivity and specificity of our HRM assay were 100%. For the blinded samples, the specificities and sensitivities were 89.3% and 100%, respectively, for detecting rifampin resistance and 98.1% and 83.3%, respectively, for detecting isoniazid resistance, as isolates with mutations in regions not encompassed by our assay were not detected. A C-to-T sequence alteration at position -15 of the ahpC regulatory region, which was previously reported to be associated with isoniazid resistance, may possibly be a polymorphism, as it was detected in an isoniazid-susceptible M. tuberculosis isolate. HRM is a rapid, accurate, simple, closed-tube, and low-cost method. It is thus an ideal assay to be used in countries with a high prevalence of drug-resistant M. tuberculosis and where cost-effectiveness is essential. As a mutation-scanning assay for detecting drug-resistant M. tuberculosis, it can potentially lead to better treatment outcomes resulting from earlier treatment with the appropriate antibiotics. Copyright © 2010, American society ror Microbiology. All Rights Reserved.
Persistent Identifierhttp://hdl.handle.net/10722/125140
ISSN
2015 Impact Factor: 3.631
2015 SCImago Journal Rankings: 2.151
PubMed Central ID
ISI Accession Number ID
Funding AgencyGrant Number
Biomedical Research Council (BMRC) of SingaporeBMRC 07/1/31/19/511
Funding Information:

This work was supported by a grant from the Biomedical Research Council (BMRC) of Singapore (BMRC 07/1/31/19/511).

References

 

DC FieldValueLanguage
dc.contributor.authorOng, DCTen_HK
dc.contributor.authorYam, WCen_HK
dc.contributor.authorSiu, GKHen_HK
dc.contributor.authorLee, ASGen_HK
dc.date.accessioned2010-10-31T11:13:40Z-
dc.date.available2010-10-31T11:13:40Z-
dc.date.issued2010en_HK
dc.identifier.citationJournal Of Clinical Microbiology, 2010, v. 48 n. 4, p. 1047-1054en_HK
dc.identifier.issn0095-1137en_HK
dc.identifier.urihttp://hdl.handle.net/10722/125140-
dc.description.abstractWe have developed a high-resolution melting (HRM) assay to scan for mutations in the rpoB, inhA, ahpC, and katG genes and/or promoter regions for the detection of rifampin and isoniazid resistance in Mycobacterium tuberculosis. For assay development, 23 drug-resistant isolates of M. tuberculosis having 29 different mutations, together with 40 drug-susceptible isolates, were utilized. All 29 mutations were accurately detected by our assay. We further validated the assay with a series of 59 samples tested in a blind manner. All sequence alterations that were within the regions targeted by the HRM assay were correctly identified. Compared against results of DNA sequencing, the sensitivity and specificity of our HRM assay were 100%. For the blinded samples, the specificities and sensitivities were 89.3% and 100%, respectively, for detecting rifampin resistance and 98.1% and 83.3%, respectively, for detecting isoniazid resistance, as isolates with mutations in regions not encompassed by our assay were not detected. A C-to-T sequence alteration at position -15 of the ahpC regulatory region, which was previously reported to be associated with isoniazid resistance, may possibly be a polymorphism, as it was detected in an isoniazid-susceptible M. tuberculosis isolate. HRM is a rapid, accurate, simple, closed-tube, and low-cost method. It is thus an ideal assay to be used in countries with a high prevalence of drug-resistant M. tuberculosis and where cost-effectiveness is essential. As a mutation-scanning assay for detecting drug-resistant M. tuberculosis, it can potentially lead to better treatment outcomes resulting from earlier treatment with the appropriate antibiotics. Copyright © 2010, American society ror Microbiology. All Rights Reserved.en_HK
dc.languageengen_HK
dc.publisherAmerican Society for Microbiology.-
dc.relation.ispartofJournal of Clinical Microbiologyen_HK
dc.rightsJournal of Clinical Microbiology. Copyright © American Society for Microbiology.-
dc.rightsCopyright © American Society for Microbiology, [insert journal name, volume number, page numbers, and year]-
dc.subject.meshAntitubercular Agents - pharmacology-
dc.subject.meshDNA, Bacterial - chemistry - genetics-
dc.subject.meshIsoniazid - pharmacology-
dc.subject.meshMycobacterium tuberculosis - drug effects - genetics - isolation and purification-
dc.subject.meshRifampin - pharmacology-
dc.titleRapid detection of rifampicin- and isoniazid-resistant mycobacterium tuberculosis by high-resolution melting analysisen_HK
dc.typeArticleen_HK
dc.identifier.openurlhttp://library.hku.hk:4550/resserv?sid=HKU:IR&issn=0095-1137&volume=48&issue=4&spage=1047&epage=1054&date=2010&atitle=Rapid+detection+of+rifampicin-+and+isoniazid-resistant+Mycobacterium+tuberculosis+by+high-resolution+melting+analysisen_HK
dc.identifier.emailYam, WC:wcyam@hkucc.hku.hken_HK
dc.identifier.authorityYam, WC=rp00313en_HK
dc.description.naturelink_to_OA_fulltext-
dc.identifier.doi10.1128/JCM.02036-09en_HK
dc.identifier.pmid20164280en_HK
dc.identifier.pmcidPMC2849564-
dc.identifier.scopuseid_2-s2.0-77950494237en_HK
dc.identifier.hkuros179789en_HK
dc.relation.referenceshttp://www.scopus.com/mlt/select.url?eid=2-s2.0-77950494237&selection=ref&src=s&origin=recordpageen_HK
dc.identifier.volume48en_HK
dc.identifier.issue4en_HK
dc.identifier.spage1047en_HK
dc.identifier.epage1054en_HK
dc.identifier.isiWOS:000276153200005-
dc.publisher.placeUnited Statesen_HK
dc.identifier.scopusauthoridOng, DCT=23486508800en_HK
dc.identifier.scopusauthoridYam, WC=7004281720en_HK
dc.identifier.scopusauthoridSiu, GKH=35485473100en_HK
dc.identifier.scopusauthoridLee, ASG=24329929200en_HK

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