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Article: Comparison of two automated DNA amplification systems with a manual one- tube nested PCR assay for diagnosis of pulmonary tuberculosis

TitleComparison of two automated DNA amplification systems with a manual one- tube nested PCR assay for diagnosis of pulmonary tuberculosis
Authors
Issue Date1997
PublisherAmerican Society for Microbiology.
Citation
Journal Of Clinical Microbiology, 1997, v. 35 n. 6, p. 1385-1389 How to Cite?
AbstractEighty-four specimens of respiratory secretions culture positive for mycobacteria (70 positive for Mycobacterium tuberculosis and 14 positive for nontuberculous mycobacteria) and 120 culture-negative specimens were evaluated by three DNA amplification techniques: a manual in-house single- tube nested PCR (nPCR) and two commercial automated assays (the Cobas Amplicor System [aPCR-h] from Roche Diagnostic Systems and the Abbott LCx Probe System [aLCx-p] from Abbott Laboratories). The overall diagnostic sensitivities of the nPCR, aPCR-h, and aLCx-p were 77.1, 84.3, and 77.1%, respectively, and the sensitivities were 57.9, 57.9, and 36.8%, respectively, for smear-negative specimens. Specimens culture positive for nontuberculous mycobacteria were negative by all three assays. Eight culture-negative specimens which were positive by one or more assays had previously been documented by culture to be positive for M. tuberculosis and were taken from patients who were treated with antituberculosis agents. Retesting of specimens negative by one assay by the other two assays revealed that each test had its unique group of negative specimens. When considering the DNA extraction and amplification steps of these assays separately, it was found that extracts from aPCR-h and aLCx-p were compatible with nPCR amplication, while the two automated assays could only amplify extracts processed with their own reagents. Limiting dilution analysis revealed that the order of analytical sensitivity was nPCR, followed by aLCx-p and then aPCR-h. Comparison of the work flow of each assay revealed that although the aPCR-h demands the least specimen handling, the turnaround time of aLCx-p is the most favorable.
Persistent Identifierhttp://hdl.handle.net/10722/49182
ISSN
2015 Impact Factor: 3.631
2015 SCImago Journal Rankings: 2.151
PubMed Central ID
ISI Accession Number ID
References

 

DC FieldValueLanguage
dc.contributor.authorYuen, KYen_HK
dc.contributor.authorYam, WCen_HK
dc.contributor.authorWong, LPOen_HK
dc.contributor.authorSeto, WHen_HK
dc.date.accessioned2008-06-12T06:36:14Z-
dc.date.available2008-06-12T06:36:14Z-
dc.date.issued1997en_HK
dc.identifier.citationJournal Of Clinical Microbiology, 1997, v. 35 n. 6, p. 1385-1389en_HK
dc.identifier.issn0095-1137en_HK
dc.identifier.urihttp://hdl.handle.net/10722/49182-
dc.description.abstractEighty-four specimens of respiratory secretions culture positive for mycobacteria (70 positive for Mycobacterium tuberculosis and 14 positive for nontuberculous mycobacteria) and 120 culture-negative specimens were evaluated by three DNA amplification techniques: a manual in-house single- tube nested PCR (nPCR) and two commercial automated assays (the Cobas Amplicor System [aPCR-h] from Roche Diagnostic Systems and the Abbott LCx Probe System [aLCx-p] from Abbott Laboratories). The overall diagnostic sensitivities of the nPCR, aPCR-h, and aLCx-p were 77.1, 84.3, and 77.1%, respectively, and the sensitivities were 57.9, 57.9, and 36.8%, respectively, for smear-negative specimens. Specimens culture positive for nontuberculous mycobacteria were negative by all three assays. Eight culture-negative specimens which were positive by one or more assays had previously been documented by culture to be positive for M. tuberculosis and were taken from patients who were treated with antituberculosis agents. Retesting of specimens negative by one assay by the other two assays revealed that each test had its unique group of negative specimens. When considering the DNA extraction and amplification steps of these assays separately, it was found that extracts from aPCR-h and aLCx-p were compatible with nPCR amplication, while the two automated assays could only amplify extracts processed with their own reagents. Limiting dilution analysis revealed that the order of analytical sensitivity was nPCR, followed by aLCx-p and then aPCR-h. Comparison of the work flow of each assay revealed that although the aPCR-h demands the least specimen handling, the turnaround time of aLCx-p is the most favorable.en_HK
dc.format.extent418 bytes-
dc.format.mimetypetext/html-
dc.languageengen_HK
dc.publisherAmerican Society for Microbiology.en_HK
dc.relation.ispartofJournal of Clinical Microbiologyen_HK
dc.rightsCreative Commons: Attribution 3.0 Hong Kong License-
dc.rightsJournal of Clinical Microbiology. Copyright © American Society for Microbiology.en_HK
dc.rightsCopyright © American Society for Microbiology, Journal of Clinical Microbiology, 1997, v. 35 n. 6, p. 1385-1389en_HK
dc.subject.meshDNA, Bacterial - analysisen_HK
dc.subject.meshLipoproteinsen_HK
dc.subject.meshNucleic Acid Amplification Techniquesen_HK
dc.subject.meshPolymerase Chain Reaction - methodsen_HK
dc.subject.meshTuberculosis, Pulmonary - diagnosis - microbiologyen_HK
dc.titleComparison of two automated DNA amplification systems with a manual one- tube nested PCR assay for diagnosis of pulmonary tuberculosisen_HK
dc.typeArticleen_HK
dc.identifier.openurlhttp://library.hku.hk:4550/resserv?sid=HKU:IR&issn=0095-1137&volume=35&issue=6&spage=1385&epage=1389&date=1997&atitle=Comparison+of+two+automated+DNA+amplification+systems+with+a+manual+one-tube+nested+PCR+assay+for+diagnosis+of+pulmonary+tuberculosisen_HK
dc.identifier.emailYuen, KY:kyyuen@hkucc.hku.hken_HK
dc.identifier.emailYam, WC:wcyam@hkucc.hku.hken_HK
dc.identifier.authorityYuen, KY=rp00366en_HK
dc.identifier.authorityYam, WC=rp00313en_HK
dc.description.naturepublished_or_final_versionen_HK
dc.identifier.pmid9163449-
dc.identifier.pmcidPMC229754-
dc.identifier.scopuseid_2-s2.0-0030978299en_HK
dc.identifier.hkuros27574-
dc.relation.referenceshttp://www.scopus.com/mlt/select.url?eid=2-s2.0-0030978299&selection=ref&src=s&origin=recordpageen_HK
dc.identifier.volume35en_HK
dc.identifier.issue6en_HK
dc.identifier.spage1385en_HK
dc.identifier.epage1389en_HK
dc.identifier.isiWOS:A1997XA75500018-
dc.publisher.placeUnited Statesen_HK
dc.identifier.scopusauthoridYuen, KY=36078079100en_HK
dc.identifier.scopusauthoridYam, WC=7004281720en_HK
dc.identifier.scopusauthoridWong, LPO=36947363200en_HK
dc.identifier.scopusauthoridSeto, WH=7005799377en_HK

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