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Title: | A Novel reverse transcriptase-polymerase chain reaction based-liquid hybridisation(RT-PCR-LH) assay for early diagnosis of dengue infection |
Authors: | Gunasekera, M.B. Hapugoda, M.D. Gunasena, S. Subasinghe, S.A.S.C. Bandara, K.B.A.T. Khan, K.B. Abeyewickreme, W. |
Keywords: | Severe Dengue Severe Dengue-diagnosis DNA, Viral-analysis Dengue Virus-isolation and purification Enzyme-Linked Immunosorbent Assay Reverse Transcriptase Polymerase Chain Reaction-methods Cohort Studies |
Issue Date: | 2003 |
Publisher: | Sri Lanka Medical Association |
Citation: | The Ceylon Medical Journal. 2003; 48(1): pp.17-22 |
Abstract: | BACKGROUND: 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 radio-labelled 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. |
Description: | Indexed in MEDLINE |
URI: | http://repository.kln.ac.lk/handle/123456789/1574 |
ISSN: | 0009-0875 (Print) |
Appears in Collections: | Journal/Magazine Articles |
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