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Article: Direct detection of rifampin-resistant Mycobacterium tuberculosis in respiratory specimens by PCR-DNA sequencing

TitleDirect detection of rifampin-resistant Mycobacterium tuberculosis in respiratory specimens by PCR-DNA sequencing
Authors
Issue Date2004
PublisherAmerican Society for Microbiology.
Citation
Journal Of Clinical Microbiology, 2004, v. 42 n. 10, p. 4438-4443 How to Cite?
AbstractThis study evaluated the feasibility of a molecular strategy based on identification of Mycobacterium tuberculosis by IS6110 PCR or Cobas Amplicor PCR, and rpoB PCR-DNA sequencing of the 81-bp rifampin resistance determining region (RRDR) for direct detection of rifampin resistance in respiratory specimens. A collection of 2,138 respiratory specimens and 352 nonduplicate M. tuberculosis isolates (including 233 isolates from the evaluated respiratory specimens and an additional collection of 119 stored isolates) from Southern China was investigated. Using culture as the reference test, the overall diagnostic sensitivities of an acid-fast bacillus (AFB) smear, Cobas Amplicor PCR, IS6110 PCR were 54.5% (156 of 286), 86.7% (248 of 286), and 89.2% (255 of 286), respectively. The sensitivities of the rpoB PCR for the specimens with positive AFB smears and with positive PCR results in the IS6110 PCR and/or Cobas Amplicor PCR were 100% (156 of 156) and 92.3% (239 of 259), respectively. Of the 352 nonduplicate M. tuberculosis isolates, the agar proportion method for rifampin reported 39 resistant strains. Full agreement (352 of 352) was found with the agar proportion method and the genotype inferred from the rpoB DNA sequencing data for rifampin. Thirty-nine mutations of nine distinct kinds, eight point mutations, and one deletion within the RRBR were found in the 39 resistant strains. For the direct DNA sequencing performed on rpoB PCR-positive respiratory specimens, the concordance with the agar proportion method and the subsequent PCR-sequencing for the culture isolate was 100%. This strategy has potential application for direct and rapid diagnosis of rifampin-resistant M. tuberculosis in IS6110 PCR or Cobas Amplicor PCR-positive respiratory specimens.
Persistent Identifierhttp://hdl.handle.net/10722/49150
ISSN
2015 Impact Factor: 3.631
2015 SCImago Journal Rankings: 2.151
PubMed Central ID
ISI Accession Number ID
References

 

DC FieldValueLanguage
dc.contributor.authorYam, WCen_HK
dc.contributor.authorTam, CMen_HK
dc.contributor.authorLeung, CCen_HK
dc.contributor.authorTong, HLen_HK
dc.contributor.authorChan, KHen_HK
dc.contributor.authorLeung, ETYen_HK
dc.contributor.authorWong, KCen_HK
dc.contributor.authorYew, WWen_HK
dc.contributor.authorSeto, WHen_HK
dc.contributor.authorYuen, KYen_HK
dc.contributor.authorHo, PLen_HK
dc.date.accessioned2008-06-12T06:35:33Z-
dc.date.available2008-06-12T06:35:33Z-
dc.date.issued2004en_HK
dc.identifier.citationJournal Of Clinical Microbiology, 2004, v. 42 n. 10, p. 4438-4443en_HK
dc.identifier.issn0095-1137en_HK
dc.identifier.urihttp://hdl.handle.net/10722/49150-
dc.description.abstractThis study evaluated the feasibility of a molecular strategy based on identification of Mycobacterium tuberculosis by IS6110 PCR or Cobas Amplicor PCR, and rpoB PCR-DNA sequencing of the 81-bp rifampin resistance determining region (RRDR) for direct detection of rifampin resistance in respiratory specimens. A collection of 2,138 respiratory specimens and 352 nonduplicate M. tuberculosis isolates (including 233 isolates from the evaluated respiratory specimens and an additional collection of 119 stored isolates) from Southern China was investigated. Using culture as the reference test, the overall diagnostic sensitivities of an acid-fast bacillus (AFB) smear, Cobas Amplicor PCR, IS6110 PCR were 54.5% (156 of 286), 86.7% (248 of 286), and 89.2% (255 of 286), respectively. The sensitivities of the rpoB PCR for the specimens with positive AFB smears and with positive PCR results in the IS6110 PCR and/or Cobas Amplicor PCR were 100% (156 of 156) and 92.3% (239 of 259), respectively. Of the 352 nonduplicate M. tuberculosis isolates, the agar proportion method for rifampin reported 39 resistant strains. Full agreement (352 of 352) was found with the agar proportion method and the genotype inferred from the rpoB DNA sequencing data for rifampin. Thirty-nine mutations of nine distinct kinds, eight point mutations, and one deletion within the RRBR were found in the 39 resistant strains. For the direct DNA sequencing performed on rpoB PCR-positive respiratory specimens, the concordance with the agar proportion method and the subsequent PCR-sequencing for the culture isolate was 100%. This strategy has potential application for direct and rapid diagnosis of rifampin-resistant M. tuberculosis in IS6110 PCR or Cobas Amplicor PCR-positive respiratory specimens.en_HK
dc.format.extent386 bytes-
dc.format.mimetypetext/html-
dc.languageengen_HK
dc.publisherAmerican Society for Microbiology.en_HK
dc.relation.ispartofJournal of Clinical Microbiologyen_HK
dc.rightsJournal of Clinical Microbiology. Copyright © American Society for Microbiology.en_HK
dc.rightsCreative Commons: Attribution 3.0 Hong Kong License-
dc.rightsCopyright © American Society for Microbiology, Journal of Clinical Microbiology, 2004, v. 42 n. 10, p. 4438-4443en_HK
dc.subject.meshAntibiotics, Antitubercular - pharmacologyen_HK
dc.subject.meshDrug Resistance, Bacterial - geneticsen_HK
dc.subject.meshMycobacterium tuberculosis - drug effects - isolation & purificationen_HK
dc.subject.meshPolymerase Chain Reaction - methodsen_HK
dc.subject.meshRifampin - pharmacologyen_HK
dc.titleDirect detection of rifampin-resistant Mycobacterium tuberculosis in respiratory specimens by PCR-DNA sequencingen_HK
dc.typeArticleen_HK
dc.identifier.openurlhttp://library.hku.hk:4550/resserv?sid=HKU:IR&issn=0095-1137&volume=42&issue=10&spage=4438&epage=4443&date=2004&atitle=Direct+detection+of+rifampin-resistant+Mycobacterium+tuberculosis+in+respiratory+specimens+by+PCR-DNA+sequencingen_HK
dc.identifier.emailYam, WC:wcyam@hkucc.hku.hken_HK
dc.identifier.emailYuen, KY:kyyuen@hkucc.hku.hken_HK
dc.identifier.emailHo, PL:plho@hkucc.hku.hken_HK
dc.identifier.authorityYam, WC=rp00313en_HK
dc.identifier.authorityYuen, KY=rp00366en_HK
dc.identifier.authorityHo, PL=rp00406en_HK
dc.description.naturepublished_or_final_versionen_HK
dc.identifier.doi10.1128/JCM.42.10.4438-4443.2004en_HK
dc.identifier.pmid15472290en_HK
dc.identifier.pmcidPMC522342en_HK
dc.identifier.scopuseid_2-s2.0-5444273459en_HK
dc.identifier.hkuros103797-
dc.relation.referenceshttp://www.scopus.com/mlt/select.url?eid=2-s2.0-5444273459&selection=ref&src=s&origin=recordpageen_HK
dc.identifier.volume42en_HK
dc.identifier.issue10en_HK
dc.identifier.spage4438en_HK
dc.identifier.epage4443en_HK
dc.identifier.isiWOS:000224473000003-
dc.publisher.placeUnited Statesen_HK
dc.identifier.scopusauthoridYam, WC=7004281720en_HK
dc.identifier.scopusauthoridTam, CM=7201442997en_HK
dc.identifier.scopusauthoridLeung, CC=7402612644en_HK
dc.identifier.scopusauthoridTong, HL=36780567800en_HK
dc.identifier.scopusauthoridChan, KH=35338760600en_HK
dc.identifier.scopusauthoridLeung, ETY=26661142200en_HK
dc.identifier.scopusauthoridWong, KC=7404759813en_HK
dc.identifier.scopusauthoridYew, WW=7005934631en_HK
dc.identifier.scopusauthoridSeto, WH=7005799377en_HK
dc.identifier.scopusauthoridYuen, KY=36078079100en_HK
dc.identifier.scopusauthoridHo, PL=7402211363en_HK

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