Teoh, B.T. and Sam, Sing Sin and Tan, K.K. and Danlami, M.B. and Shu, M.H. and Johari, Jefree and Hooi, P.S. and Brooks, D. and Piepenburg, O. and Nentwich, O. and Wilder-Smith, A. and Franco, L. and Tenorio, A. and AbuBakar, Sazaly (2015) Early Detection of Dengue Virus by Use of Reverse Transcription-Recombinase Polymerase Amplification. Journal of Clinical Microbiology, 53 (3). pp. 830-837. ISSN 0095-1137, DOI https://doi.org/10.1128/JCM.02648-14.
Full text not available from this repository.Abstract
A method for the rapid diagnosis of early dengue virus (DENV) infection is highly needed. Here, a prototype reverse transcriptionrecombinase polymerase amplification (RT-RPA) assay was developed. The assay detected DENV RNA in <20 min without the need for thermocycling amplification. The assay enabled the detection of as few as 10 copies of DENV RNA. The designed RT-RPA primers and exo probe detected the DENV genome of at least 12 genotypes of DENV circulating globally without crossreacting with other arboviruses. We assessed the diagnostic performance of the RT-RPA assay for the detection of DENV RNA in 203 serum samples of patients with clinically suspected dengue. The sera were simultaneously tested for DENV using a reverse transcription-loop-mediated isothermal amplification (RT-LAMP) assay, quantitative RT-PCR (qRT-PCR), and IgM- and IgGcapture enzyme-linked immunosorbent assays (ELISA). Acute DENV infection was confirmed in 130 samples and 61 of the samples (46.9%) were classified as viremic with qRT-PCR. The RT-RPA assay showed good concordance (κ of ≥0.723) with the RTLAMP and qRT-PCR assays in detecting the dengue viremic samples. When used in combination with ELISA, both the RT-RPA and RT-LAMP assays increased the detection of acute DENV infection to ≥95.7% (≥45/47) in samples obtained within 5 days of illness. The results from the study suggest that the RT-RPA assay is the most rapid molecular diagnostic tool available for the detection of DENV. Hence, it is possible to use the RT-RPA assay in a laboratory to complement routine serology testing for dengue.
| Item Type: | Article |
|---|---|
| Funders: | European Union's Seventh Framework Programme (FP7/2007-2013) under grant agreement 282589 (DengueTools) |
| Uncontrolled Keywords: | Adolescent; Adult; Aged; Child; Child, Preschool; Cohort Studies; Dengue; Dengue Virus; Early Diagnosis; Female; Genotype; Humans; Male; Middle Aged; Molecular Diagnostic Techniques; Reverse Transcriptase Polymerase Chain Reaction; RNA, Viral; Sensitivity and Specificity; Time Factors; Young Adult |
| Subjects: | R Medicine |
| Divisions: | Faculty of Medicine |
| Depositing User: | Ms. Juhaida Abd Rahim |
| Date Deposited: | 03 Oct 2018 05:31 |
| Last Modified: | 13 Feb 2019 08:13 |
| URI: | http://eprints.um.edu.my/id/eprint/19565 |
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