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Article: Diagnostic evaluation of an in-house developed single-tube, duplex, nested IS6110 real-time PCR assay for rapid pulmonary tuberculosis diagnosis

TitleDiagnostic evaluation of an in-house developed single-tube, duplex, nested IS6110 real-time PCR assay for rapid pulmonary tuberculosis diagnosis
Authors
KeywordsMycobacterium tuberculosis
Diagnostic
IS6110
Real-time PCR
Issue Date2018
PublisherElsevier Ltd. The Journal's web site is located at http://www.elsevier.com/locate/tube
Citation
Tuberculosis, 2018, v. 112, p. 120-125 How to Cite?
AbstractObjective: To perform a prospective evaluation on the diagnostic performance of an in-house developed, duplex nested IS6110 real-time Polymerase-Chain-Reaction (PCR) assay (IS6110-qPCR assay) for rapid pulmonary TB diagnosis. Methods: A total of 503 sputum specimens were prospectively collected from July 2016 to November 2016. Diagnostic accuracy and optimal cut-off Cycle-threshold (Ct) value for IS6110-qPCR assay was determined by Receiver Operating Characteristic (ROC) curve. Using the optimal cut-off Ct, diagnostic performance of IS6110-qPCR assay was assessed with reference to both bacteriological and clinical information. Meanwhile, limit of detection (LOD) was calculated using Mycobacterium tuberculosis H37Rv as reference strain. Result: ROC curve analysis of IS6110-qPCR assay showed a high Area Under the Curve (AUC) value (0.948) with optimal Ct value at 24.140. Prospective analysis of IS6110-qPCR assay with cut-off Ct = 24.140 showed a high overall sensitivity and specificity of 97.2% and 99.7%, respectively. No cross reactivity was observed among all non-tuberculous mycobacteria specimens in this study. LOD analysis on MTB-spiked sputum showed an average detection limit of 5.0 CFU/mL at Ct = 23.18 (±SD, 0.57). Conclusion: IS6110-qPCR assay is a highly accurate and cost-effective assay developed for primary screening of suspected TB cases, which is particularly suitable for regions with limited resources but high TB burden.
Persistent Identifierhttp://hdl.handle.net/10722/285300
ISSN
2019 Impact Factor: 2.576
2015 SCImago Journal Rankings: 1.504
ISI Accession Number ID

 

DC FieldValueLanguage
dc.contributor.authorLEUNG, KSS-
dc.contributor.authorSIU, GKH-
dc.contributor.authorTAM, KGK-
dc.contributor.authorHo, PL-
dc.contributor.authorWong, SSY-
dc.contributor.authorLeung, EKC-
dc.contributor.authorYu, SH-
dc.contributor.authorMa, OCK-
dc.contributor.authorYam, WC-
dc.date.accessioned2020-08-18T03:52:10Z-
dc.date.available2020-08-18T03:52:10Z-
dc.date.issued2018-
dc.identifier.citationTuberculosis, 2018, v. 112, p. 120-125-
dc.identifier.issn1472-9792-
dc.identifier.urihttp://hdl.handle.net/10722/285300-
dc.description.abstractObjective: To perform a prospective evaluation on the diagnostic performance of an in-house developed, duplex nested IS6110 real-time Polymerase-Chain-Reaction (PCR) assay (IS6110-qPCR assay) for rapid pulmonary TB diagnosis. Methods: A total of 503 sputum specimens were prospectively collected from July 2016 to November 2016. Diagnostic accuracy and optimal cut-off Cycle-threshold (Ct) value for IS6110-qPCR assay was determined by Receiver Operating Characteristic (ROC) curve. Using the optimal cut-off Ct, diagnostic performance of IS6110-qPCR assay was assessed with reference to both bacteriological and clinical information. Meanwhile, limit of detection (LOD) was calculated using Mycobacterium tuberculosis H37Rv as reference strain. Result: ROC curve analysis of IS6110-qPCR assay showed a high Area Under the Curve (AUC) value (0.948) with optimal Ct value at 24.140. Prospective analysis of IS6110-qPCR assay with cut-off Ct = 24.140 showed a high overall sensitivity and specificity of 97.2% and 99.7%, respectively. No cross reactivity was observed among all non-tuberculous mycobacteria specimens in this study. LOD analysis on MTB-spiked sputum showed an average detection limit of 5.0 CFU/mL at Ct = 23.18 (±SD, 0.57). Conclusion: IS6110-qPCR assay is a highly accurate and cost-effective assay developed for primary screening of suspected TB cases, which is particularly suitable for regions with limited resources but high TB burden.-
dc.languageeng-
dc.publisherElsevier Ltd. The Journal's web site is located at http://www.elsevier.com/locate/tube-
dc.relation.ispartofTuberculosis-
dc.subjectMycobacterium tuberculosis-
dc.subjectDiagnostic-
dc.subjectIS6110-
dc.subjectReal-time PCR-
dc.titleDiagnostic evaluation of an in-house developed single-tube, duplex, nested IS6110 real-time PCR assay for rapid pulmonary tuberculosis diagnosis-
dc.typeArticle-
dc.identifier.emailHo, PL: plho@hku.hk-
dc.identifier.emailWong, SSY: samsonsy@hku.hk-
dc.identifier.emailLeung, EKC: edgrmhe@hku.hk-
dc.identifier.emailYam, WC: wcyam@hku.hk-
dc.identifier.authorityHo, PL=rp00406-
dc.identifier.authorityWong, SSY=rp00395-
dc.identifier.authorityYam, WC=rp00313-
dc.description.naturelink_to_subscribed_fulltext-
dc.description.naturelink_to_subscribed_fulltext-
dc.identifier.doi10.1016/j.tube.2018.08.008-
dc.identifier.pmid30205964-
dc.identifier.scopuseid_2-s2.0-85052461846-
dc.identifier.scopuseid_2-s2.0-85052461846-
dc.identifier.hkuros312819-
dc.identifier.volume112-
dc.identifier.spage120-
dc.identifier.epage125-
dc.identifier.isiWOS:000443991700016-
dc.publisher.placeUnited Kingdom-
dc.identifier.issnl1472-9792-

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