File Download
  Links for fulltext
     (May Require Subscription)
Supplementary

Article: Single-tube nested PCR in the diagnosis of tuberculosis

TitleSingle-tube nested PCR in the diagnosis of tuberculosis
Authors
KeywordsNested PCR
Tuberculosis
Uracil-N-glycosylase
Issue Date1996
PublisherB M J Publishing Group. The Journal's web site is located at http://jcp.bmjjournals.com/
Citation
Journal Of Clinical Pathology, 1996, v. 49 n. 4, p. 290-294 How to Cite?
AbstractAims - To evaluate the usefulness of a single-tube nested polymerase chain reaction (PCR) assay in the diagnosis of tuberculosis in 1497 pulmonary and 536 extrapulmonary specimens. Methods-A single-tube nested PCR, utilising two sets of primers with different melting temperatures (88°C for external primers; 70°C for internal primers) to augment sensitivity and specificity without increasing the risk of amplicon contamination, was evaluated. Specimens were initially tested for the repetitive IS6110 sequences and if negative, retested for the universal 38 kilodalton sequence and for inhibitors. dUTP/Uracil-N-glycosylase and Instagene treatment were used to minimise contamination and the effect of inhibitors, respectively. Results - Using culture as the gold standard, the overall sensitivity of the assay was 89% for pulmonary and 42% for extrapulmonary specimens. Sensitivity varied greatly with respect to sample type (92% for follow up specimens from a chest hospital and 70% for non-follow up specimens from a general hospital). The smear positivity rates were 15% for extrapulmonary specimens, and 69% and 45%, respectively, for follow up and non-follow up specimens from pulmonary sites. Specificity was 99.7%. Inhibitors were present more frequently in extrapulmonary than in pulmonary specimens (13.4% v 2.7%). Conclusion - Despite the high sensitivity of the PCR assay for the diagnosis of tuberculosis in pulmonary specimens, it was less effective in the extrapulmonary samples. This is probably because of the lower bacterial load in extrapulmonary specimens, the presence of more inhibitors adversely affecting the PCR assay and the higher volume of specimens used for culture.
Persistent Identifierhttp://hdl.handle.net/10722/43120
ISSN
2015 Impact Factor: 2.912
2015 SCImago Journal Rankings: 1.260
PubMed Central ID
ISI Accession Number ID
References

 

DC FieldValueLanguage
dc.contributor.authorChan, CMen_HK
dc.contributor.authorYuen, KYen_HK
dc.contributor.authorChan, KSen_HK
dc.contributor.authorYam, WCen_HK
dc.contributor.authorYim, KHMen_HK
dc.contributor.authorNg, WFen_HK
dc.contributor.authorNg, MHen_HK
dc.date.accessioned2007-03-23T04:39:21Z-
dc.date.available2007-03-23T04:39:21Z-
dc.date.issued1996en_HK
dc.identifier.citationJournal Of Clinical Pathology, 1996, v. 49 n. 4, p. 290-294en_HK
dc.identifier.issn0021-9746en_HK
dc.identifier.urihttp://hdl.handle.net/10722/43120-
dc.description.abstractAims - To evaluate the usefulness of a single-tube nested polymerase chain reaction (PCR) assay in the diagnosis of tuberculosis in 1497 pulmonary and 536 extrapulmonary specimens. Methods-A single-tube nested PCR, utilising two sets of primers with different melting temperatures (88°C for external primers; 70°C for internal primers) to augment sensitivity and specificity without increasing the risk of amplicon contamination, was evaluated. Specimens were initially tested for the repetitive IS6110 sequences and if negative, retested for the universal 38 kilodalton sequence and for inhibitors. dUTP/Uracil-N-glycosylase and Instagene treatment were used to minimise contamination and the effect of inhibitors, respectively. Results - Using culture as the gold standard, the overall sensitivity of the assay was 89% for pulmonary and 42% for extrapulmonary specimens. Sensitivity varied greatly with respect to sample type (92% for follow up specimens from a chest hospital and 70% for non-follow up specimens from a general hospital). The smear positivity rates were 15% for extrapulmonary specimens, and 69% and 45%, respectively, for follow up and non-follow up specimens from pulmonary sites. Specificity was 99.7%. Inhibitors were present more frequently in extrapulmonary than in pulmonary specimens (13.4% v 2.7%). Conclusion - Despite the high sensitivity of the PCR assay for the diagnosis of tuberculosis in pulmonary specimens, it was less effective in the extrapulmonary samples. This is probably because of the lower bacterial load in extrapulmonary specimens, the presence of more inhibitors adversely affecting the PCR assay and the higher volume of specimens used for culture.en_HK
dc.format.extent1169700 bytes-
dc.format.extent30720 bytes-
dc.format.extent3548 bytes-
dc.format.mimetypeapplication/pdf-
dc.format.mimetypeapplication/msword-
dc.format.mimetypetext/plain-
dc.languageengen_HK
dc.publisherB M J Publishing Group. The Journal's web site is located at http://jcp.bmjjournals.com/en_HK
dc.relation.ispartofJournal of Clinical Pathologyen_HK
dc.rightsJournal of Clinical Pathology. Copyright © B M J Publishing Group.en_HK
dc.rightsCreative Commons: Attribution 3.0 Hong Kong License-
dc.subjectNested PCRen_HK
dc.subjectTuberculosisen_HK
dc.subjectUracil-N-glycosylaseen_HK
dc.subject.meshPolymerase chain reaction -methodsen_HK
dc.subject.meshTuberculosis, pulmonary - diagnosisen_HK
dc.subject.meshSensitivity and specificityen_HK
dc.subject.meshBlotting, southernen_HK
dc.subject.meshBase sequenceen_HK
dc.titleSingle-tube nested PCR in the diagnosis of tuberculosisen_HK
dc.typeArticleen_HK
dc.identifier.openurlhttp://library.hku.hk:4550/resserv?sid=HKU:IR&issn=0021-9746&volume=49&issue=4&spage=290&epage=294&date=1996&atitle=Single-tube+nested+PCR+in+the+diagnosis+of+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.pmid8655703en_HK
dc.identifier.pmcidPMC500452-
dc.identifier.scopuseid_2-s2.0-0029881253en_HK
dc.identifier.hkuros14579-
dc.relation.referenceshttp://www.scopus.com/mlt/select.url?eid=2-s2.0-0029881253&selection=ref&src=s&origin=recordpageen_HK
dc.identifier.volume49en_HK
dc.identifier.issue4en_HK
dc.identifier.spage290en_HK
dc.identifier.epage294en_HK
dc.identifier.isiWOS:A1996UG22800005-
dc.publisher.placeUnited Kingdomen_HK
dc.identifier.scopusauthoridChan, CM=7404814453en_HK
dc.identifier.scopusauthoridYuen, KY=36078079100en_HK
dc.identifier.scopusauthoridChan, KS=36448818400en_HK
dc.identifier.scopusauthoridYam, WC=7004281720en_HK
dc.identifier.scopusauthoridYim, KHM=7005983613en_HK
dc.identifier.scopusauthoridNg, WF=55138043100en_HK
dc.identifier.scopusauthoridNg, MH=7202076421en_HK

Export via OAI-PMH Interface in XML Formats


OR


Export to Other Non-XML Formats