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Article: Direct detection of isoniazid-resistant Mycobacterium tuberculosis in respiratory specimens by multiplex allele-specific polymerase chain reaction

TitleDirect detection of isoniazid-resistant Mycobacterium tuberculosis in respiratory specimens by multiplex allele-specific polymerase chain reaction
Authors
Issue Date2011
PublisherElsevier Inc. The Journal's web site is located at http://www.elsevier.com/locate/diagmicrobio
Citation
Diagnostic Microbiology And Infectious Disease, 2011, v. 69 n. 1, p. 51-58 How to Cite?
AbstractThis study evaluated the feasibility of using 2 multiplex allele-specific polymerase chain reaction (MAS-PCR) assays targeting 2 mutations (codon 315 of the katG gene and the 15th nucleotide preceding the mabA-inhA operon) to directly detect isoniazid (INH)-resistant Mycobacterium tuberculosis in cultured isolates and respiratory specimens. A total of 203 M. tuberculosis isolates and 487 respiratory specimens were investigated. The MAS-PCR assays successfully amplified all M. tuberculosis isolates and acid-fast bacilli smear-positive specimens while only 49.2% of the smear-negative specimens exhibited positive MAS-PCR results. The MAS-PCR assays identified 83.4% and 79.2% of the resistant strains in the culture isolates and respiratory specimens, respectively. All the inferred genotypes were in complete accordance with subsequent DNA sequence analyses. This study suggested the application of our improved MAS-PCR protocols to provide the rapid identification of INH-resistant M. tuberculosis directly in respiratory specimens. The technical simplicity, short turnaround time, and low cost of this molecular strategy should facilitate routine diagnostic services in developing areas with a high prevalence of drug-resistant tuberculosis. © 2011 Elsevier Inc.
Persistent Identifierhttp://hdl.handle.net/10722/157610
ISSN
2015 Impact Factor: 2.45
2015 SCImago Journal Rankings: 1.142
ISI Accession Number ID
Funding AgencyGrant Number
Health, Welfare and Food Bureau of the Hong Kong SAR Government04050032
Funding Information:

The work was supported by a research grant from the Research Fund for the Control of Infectious Diseases (RFCID) of the Health, Welfare and Food Bureau of the Hong Kong SAR Government (project no. 04050032).

References
Grants

 

DC FieldValueLanguage
dc.contributor.authorSiu, GKHen_US
dc.contributor.authorTam, YHen_US
dc.contributor.authorHo, PLen_US
dc.contributor.authorLee, ASGen_US
dc.contributor.authorQue, TLen_US
dc.contributor.authorTse, CWSen_US
dc.contributor.authorYip, KTen_US
dc.contributor.authorLam, JTHen_US
dc.contributor.authorCheng, VCCen_US
dc.contributor.authorYuen, KYen_US
dc.contributor.authorYam, WCen_US
dc.date.accessioned2012-08-08T08:51:40Z-
dc.date.available2012-08-08T08:51:40Z-
dc.date.issued2011en_US
dc.identifier.citationDiagnostic Microbiology And Infectious Disease, 2011, v. 69 n. 1, p. 51-58en_US
dc.identifier.issn0732-8893en_US
dc.identifier.urihttp://hdl.handle.net/10722/157610-
dc.description.abstractThis study evaluated the feasibility of using 2 multiplex allele-specific polymerase chain reaction (MAS-PCR) assays targeting 2 mutations (codon 315 of the katG gene and the 15th nucleotide preceding the mabA-inhA operon) to directly detect isoniazid (INH)-resistant Mycobacterium tuberculosis in cultured isolates and respiratory specimens. A total of 203 M. tuberculosis isolates and 487 respiratory specimens were investigated. The MAS-PCR assays successfully amplified all M. tuberculosis isolates and acid-fast bacilli smear-positive specimens while only 49.2% of the smear-negative specimens exhibited positive MAS-PCR results. The MAS-PCR assays identified 83.4% and 79.2% of the resistant strains in the culture isolates and respiratory specimens, respectively. All the inferred genotypes were in complete accordance with subsequent DNA sequence analyses. This study suggested the application of our improved MAS-PCR protocols to provide the rapid identification of INH-resistant M. tuberculosis directly in respiratory specimens. The technical simplicity, short turnaround time, and low cost of this molecular strategy should facilitate routine diagnostic services in developing areas with a high prevalence of drug-resistant tuberculosis. © 2011 Elsevier Inc.en_US
dc.languageengen_US
dc.publisherElsevier Inc. The Journal's web site is located at http://www.elsevier.com/locate/diagmicrobioen_US
dc.relation.ispartofDiagnostic Microbiology and Infectious Diseaseen_US
dc.subject.meshAllelesen_US
dc.subject.meshAmino Acid Substitution - Geneticsen_US
dc.subject.meshBacterial Proteins - Geneticsen_US
dc.subject.meshCatalase - Geneticsen_US
dc.subject.meshDna, Bacterial - Geneticsen_US
dc.subject.meshDrug Resistance, Bacterialen_US
dc.subject.meshHumansen_US
dc.subject.meshIsoniazid - Pharmacologyen_US
dc.subject.meshMicrobial Sensitivity Tests - Methodsen_US
dc.subject.meshMutation, Missenseen_US
dc.subject.meshMycobacterium Tuberculosis - Drug Effects - Genetics - Isolation & Purificationen_US
dc.subject.meshOxidoreductases - Geneticsen_US
dc.subject.meshPolymerase Chain Reaction - Methodsen_US
dc.subject.meshSputum - Microbiologyen_US
dc.subject.meshTime Factorsen_US
dc.subject.meshTuberculosis - Microbiologyen_US
dc.titleDirect detection of isoniazid-resistant Mycobacterium tuberculosis in respiratory specimens by multiplex allele-specific polymerase chain reactionen_US
dc.typeArticleen_US
dc.identifier.emailHo, PL:plho@hkucc.hku.hken_US
dc.identifier.emailYuen, KY:kyyuen@hkucc.hku.hken_US
dc.identifier.emailYam, WC:wcyam@hkucc.hku.hken_US
dc.identifier.authorityHo, PL=rp00406en_US
dc.identifier.authorityYuen, KY=rp00366en_US
dc.identifier.authorityYam, WC=rp00313en_US
dc.description.naturelink_to_subscribed_fulltexten_US
dc.identifier.doi10.1016/j.diagmicrobio.2010.08.021en_US
dc.identifier.pmid21146714-
dc.identifier.scopuseid_2-s2.0-78649958683en_US
dc.relation.referenceshttp://www.scopus.com/mlt/select.url?eid=2-s2.0-78649958683&selection=ref&src=s&origin=recordpageen_US
dc.identifier.volume69en_US
dc.identifier.issue1en_US
dc.identifier.spage51en_US
dc.identifier.epage58en_US
dc.identifier.isiWOS:000285570500008-
dc.publisher.placeUnited Statesen_US
dc.relation.projectEvaluation of molecular tests for the rapid detection of rifampicin and isoniazid resistance in Mycobacterium tuberculosis and their impact on patient management-
dc.identifier.scopusauthoridSiu, GKH=35485473100en_US
dc.identifier.scopusauthoridTam, YH=37057992700en_US
dc.identifier.scopusauthoridHo, PL=7402211363en_US
dc.identifier.scopusauthoridLee, ASG=24329929200en_US
dc.identifier.scopusauthoridQue, TL=7003786628en_US
dc.identifier.scopusauthoridTse, CWS=7103295064en_US
dc.identifier.scopusauthoridYip, KT=7101909925en_US
dc.identifier.scopusauthoridLam, JTH=22941763000en_US
dc.identifier.scopusauthoridCheng, VCC=23670479400en_US
dc.identifier.scopusauthoridYuen, KY=36078079100en_US
dc.identifier.scopusauthoridYam, WC=7004281720en_US
dc.identifier.citeulike8447788-

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