File Download

There are no files associated with this item.

  Links for fulltext
     (May Require Subscription)
Supplementary

Article: Direct detection of Mycobacterium tuberculosis in clinical specimens using single-tube biotinylated nested polymerase chain reaction-enzyme linked immunoassay (PCR-ELISA)

TitleDirect detection of Mycobacterium tuberculosis in clinical specimens using single-tube biotinylated nested polymerase chain reaction-enzyme linked immunoassay (PCR-ELISA)
Authors
Issue Date2004
PublisherElsevier Inc. The Journal's web site is located at http://www.elsevier.com/locate/diagmicrobio
Citation
Diagnostic Microbiology And Infectious Disease, 2004, v. 48 n. 4, p. 271-275 How to Cite?
AbstractA biotinylated single-tube nested polymerase chain reaction (PCR) assay with microwell hybridization assay (bPCR-ELISA) was developed for detection of Mycobacterium tuberculosis in clinical specimens. A total of 659 specimens (601 respiratory specimens and 58 nonrespiratory specimens) were collected for evaluation using three DNA amplification techniques: newly designed bPCR-ELISA, in-house single-tube nested PCR for IS6110 gene sequence (nPCR), and commercial automated assays, the Cobas Amplicor System from Roche Diagnostic Systems (aPCR). Sixty-four (9.7%) specimens were culture-positive for M. tuberculosis. Eleven (1.7%) specimens culture-positive for nontuberculosis mycobacteria were negative by all three PCR assays. The resolved performance of bPCR-ELISA, nPCR, and aPCR was found at sensitivities of 97%, 94%, and 97%, respectively. All three PCR assays exhibited a 100% specificity. In evaluation of bPCR-ELISA, a clear distinction between PCR-positive and PCR-negative specimens when an OD405 value of 0.6 was chosen as cut-off. With serial dilutions of M. tuberculosis H37Rv DNA, the detection limit of bPCR-ELISA was found to be 0.75 cfu per reaction at OD405 value of 0.6. Our developed bPCR-ELISA provides a highly sensitive and low-costing molecular diagnosis suitable for developing countries with high prevalence of tuberculosis. © 2004 Elsevier Inc. All rights reserved.
Persistent Identifierhttp://hdl.handle.net/10722/157405
ISSN
2015 Impact Factor: 2.45
2015 SCImago Journal Rankings: 1.142
ISI Accession Number ID
References

 

DC FieldValueLanguage
dc.contributor.authorYam, WCen_US
dc.contributor.authorCheng, VCCen_US
dc.contributor.authorHui, WTen_US
dc.contributor.authorWang, LNen_US
dc.contributor.authorSeto, WHen_US
dc.contributor.authorYuen, KYen_US
dc.date.accessioned2012-08-08T08:49:41Z-
dc.date.available2012-08-08T08:49:41Z-
dc.date.issued2004en_US
dc.identifier.citationDiagnostic Microbiology And Infectious Disease, 2004, v. 48 n. 4, p. 271-275en_US
dc.identifier.issn0732-8893en_US
dc.identifier.urihttp://hdl.handle.net/10722/157405-
dc.description.abstractA biotinylated single-tube nested polymerase chain reaction (PCR) assay with microwell hybridization assay (bPCR-ELISA) was developed for detection of Mycobacterium tuberculosis in clinical specimens. A total of 659 specimens (601 respiratory specimens and 58 nonrespiratory specimens) were collected for evaluation using three DNA amplification techniques: newly designed bPCR-ELISA, in-house single-tube nested PCR for IS6110 gene sequence (nPCR), and commercial automated assays, the Cobas Amplicor System from Roche Diagnostic Systems (aPCR). Sixty-four (9.7%) specimens were culture-positive for M. tuberculosis. Eleven (1.7%) specimens culture-positive for nontuberculosis mycobacteria were negative by all three PCR assays. The resolved performance of bPCR-ELISA, nPCR, and aPCR was found at sensitivities of 97%, 94%, and 97%, respectively. All three PCR assays exhibited a 100% specificity. In evaluation of bPCR-ELISA, a clear distinction between PCR-positive and PCR-negative specimens when an OD405 value of 0.6 was chosen as cut-off. With serial dilutions of M. tuberculosis H37Rv DNA, the detection limit of bPCR-ELISA was found to be 0.75 cfu per reaction at OD405 value of 0.6. Our developed bPCR-ELISA provides a highly sensitive and low-costing molecular diagnosis suitable for developing countries with high prevalence of tuberculosis. © 2004 Elsevier Inc. All rights reserved.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.rightsDiagnostic Microbiology and Infectious Disease. Copyright © Elsevier Inc.-
dc.subject.meshBacteriological Techniquesen_US
dc.subject.meshBiotin - Metabolismen_US
dc.subject.meshEnzyme-Linked Immunosorbent Assayen_US
dc.subject.meshHumansen_US
dc.subject.meshMycobacterium Tuberculosis - Genetics - Isolation & Purificationen_US
dc.subject.meshPolymerase Chain Reaction - Methodsen_US
dc.subject.meshSensitivity And Specificityen_US
dc.subject.meshTuberculosis - Diagnosis - Microbiologyen_US
dc.titleDirect detection of Mycobacterium tuberculosis in clinical specimens using single-tube biotinylated nested polymerase chain reaction-enzyme linked immunoassay (PCR-ELISA)en_US
dc.typeArticleen_US
dc.identifier.emailYam, WC:wcyam@hkucc.hku.hken_US
dc.identifier.emailYuen, KY:kyyuen@hkucc.hku.hken_US
dc.identifier.authorityYam, WC=rp00313en_US
dc.identifier.authorityYuen, KY=rp00366en_US
dc.description.naturelink_to_subscribed_fulltexten_US
dc.identifier.doi10.1016/j.diagmicrobio.2003.11.006en_US
dc.identifier.pmid15062920-
dc.identifier.scopuseid_2-s2.0-1942488391en_US
dc.identifier.hkuros87950-
dc.relation.referenceshttp://www.scopus.com/mlt/select.url?eid=2-s2.0-1942488391&selection=ref&src=s&origin=recordpageen_US
dc.identifier.volume48en_US
dc.identifier.issue4en_US
dc.identifier.spage271en_US
dc.identifier.epage275en_US
dc.identifier.isiWOS:000220907300008-
dc.publisher.placeUnited Statesen_US
dc.identifier.scopusauthoridYam, WC=7004281720en_US
dc.identifier.scopusauthoridCheng, VCC=23670479400en_US
dc.identifier.scopusauthoridHui, WT=7103196480en_US
dc.identifier.scopusauthoridWang, LN=7409181161en_US
dc.identifier.scopusauthoridSeto, WH=7005799377en_US
dc.identifier.scopusauthoridYuen, KY=36078079100en_US

Export via OAI-PMH Interface in XML Formats


OR


Export to Other Non-XML Formats