A novel reverse transcriptase-polymerase chain reaction based-liquid hybridisation (RT-PCR-LH) assay for early diagnosis of dengue infection.

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dc.contributor.authorGunesekera, Maya Ben_US
dc.contributor.authorHapugoda, Menaka Den_US
dc.contributor.authorGunasena, Sen_US
dc.contributor.authorSubasinghe, S A S Cen_US
dc.contributor.authorBandara, K B A Ten_US
dc.contributor.authorKhan, Baldip Ken_US
dc.contributor.authorAbeyewickreme, Wen_US
dc.date.accessioned2009-05-28T05:16:01Z
dc.date.available2009-05-28T05:16:01Z
dc.date.issued2003-03-11en_US
dc.descriptionThe Ceylon Medical Journal.en_US
dc.description.abstractBACKGROUND: Early definitive laboratory diagnosis of dengue is difficult with the tests in routine use at present. OBJECTIVE: To develop a reverse transcriptase-polymerase chain reaction based liquid hybridisation (RT-PCR-LH) technique for the rapid and early diagnosis of dengue. RESEARCH DESIGN: RT-PCR products of the NS3 gene of dengue virus prototypes and of a few positive sera for dengue virus by culture, were allowed to hybridise in liquid phase with a mixture of dengue specific radiolabelled oligonucleotides. The products were separated by PAGE and visualised by autoradiography. 78 suspected dengue sera were also tested by RT-PCR-LH method, and by IgM-ELISA and HAI tests, for comparison. RESULTS: Two DNA bands (approximately equal to 470 bp and approximately equal to 455 bp) specific to dengue virus, were observed. RT-PCR-LH assay takes only 24 h. Of the 78 suspected dengue acute sera tested, 45/78 were positive by RT-PCR-LH, 31/78 were positive by IgM-ELISA, and 14/78 had a HAI titre > or = 2560. Duration of fever was known in 72 cases, and infection was detected by RT-PCR-LH in 11/22 of cases with < 5 d fever and by IgM-ELISA in 1/22. In cases with 5 to 15 d fever RT-PCR-LH and IgM-ELISA/HAI titre > or = 2560 detected infection in 30/50 and 27/50 respectively. The 10 sera which were negative by RT-PCR-LH, but were positive by either IgM-ELISA or HAI titre > or = 2560 were all > 5 d fever cases. RT-PCR-LH together with IgM-ELISA were capable of detecting dengue infection in 56/78 of the suspected cases. CONCLUSION: RT-PCR-LH assay developed in this study appears to have an advantage over other diagnostic techniques for the early detection of dengue.en_US
dc.description.affiliationDepartment of Chemistry, Faculty of Science, University of Colombo, Colombo 3. biotech@slt.lken_US
dc.identifier.citationGunesekera MB, Hapugoda MD, Gunasena S, Subasinghe SA, Bandara KB, Khan BK, Abeyewickreme W. A novel reverse transcriptase-polymerase chain reaction based-liquid hybridisation (RT-PCR-LH) assay for early diagnosis of dengue infection. The Ceylon Medical Journal. 2003 Mar; 48(1): 17-22en_US
dc.identifier.urihttps://imsear.searo.who.int/handle/123456789/48202
dc.language.isoengen_US
dc.source.urihttps://www.infolanka.com/CMJhome/en_US
dc.subject.meshCohort Studiesen_US
dc.subject.meshDNA, Viral --analysisen_US
dc.subject.meshDengue Hemorrhagic Fever --diagnosisen_US
dc.subject.meshDengue Virus --isolation & purificationen_US
dc.subject.meshEnzyme-Linked Immunosorbent Assayen_US
dc.subject.meshFemaleen_US
dc.subject.meshHumansen_US
dc.subject.meshMaleen_US
dc.subject.meshReverse Transcriptase Polymerase Chain Reaction --methodsen_US
dc.subject.meshSensitivity and Specificityen_US
dc.titleA novel reverse transcriptase-polymerase chain reaction based-liquid hybridisation (RT-PCR-LH) assay for early diagnosis of dengue infection.en_US
dc.typeComparative Studyen_US
dc.typeJournal Articleen_US
dc.typeResearch Support, Non-U.S. Gov'ten_US
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