Early detection of dengue virus by use of reverse transcription-recombinase polymerase amplification.

Autor:	Teoh BT; Tropical Infectious Diseases Research and Education Centre (TIDREC), Department of Medical Microbiology, Faculty of Medicine, University of Malaya, Kuala Lumpur, Malaysia., Sam SS; Tropical Infectious Diseases Research and Education Centre (TIDREC), Department of Medical Microbiology, Faculty of Medicine, University of Malaya, Kuala Lumpur, Malaysia., Tan KK; Tropical Infectious Diseases Research and Education Centre (TIDREC), Department of Medical Microbiology, Faculty of Medicine, University of Malaya, Kuala Lumpur, Malaysia., Danlami MB; Tropical Infectious Diseases Research and Education Centre (TIDREC), Department of Medical Microbiology, Faculty of Medicine, University of Malaya, Kuala Lumpur, Malaysia., Shu MH; Tropical Infectious Diseases Research and Education Centre (TIDREC), Department of Medical Microbiology, Faculty of Medicine, University of Malaya, Kuala Lumpur, Malaysia., Johari J; Tropical Infectious Diseases Research and Education Centre (TIDREC), Department of Medical Microbiology, Faculty of Medicine, University of Malaya, Kuala Lumpur, Malaysia., Hooi PS; Tropical Infectious Diseases Research and Education Centre (TIDREC), Department of Medical Microbiology, Faculty of Medicine, University of Malaya, Kuala Lumpur, Malaysia., Brooks D; TwistDx, Ltd., Minerva Building, Babraham Research Campus, Cambridge, United Kingdom., Piepenburg O; TwistDx, Ltd., Minerva Building, Babraham Research Campus, Cambridge, United Kingdom., Nentwich O; TwistDx, Ltd., Minerva Building, Babraham Research Campus, Cambridge, United Kingdom., Wilder-Smith A; Department of Public Health and Clinical Medicine, Epidemiology and Global Health, Umeå University, Umeå, Sweden Lee Kong Chian School of Medicine, Nanyang Technological University, Singapore., Franco L; Arbovirus and Imported Viral Diseases, National Centre for Microbiology, Institute of Health Carlos III, Madrid, Spain., Tenorio A; Arbovirus and Imported Viral Diseases, National Centre for Microbiology, Institute of Health Carlos III, Madrid, Spain., AbuBakar S; Tropical Infectious Diseases Research and Education Centre (TIDREC), Department of Medical Microbiology, Faculty of Medicine, University of Malaya, Kuala Lumpur, Malaysia sazaly@um.edu.my.
Jazyk:	angličtina
Zdroj:	Journal of clinical microbiology [J Clin Microbiol] 2015 Mar; Vol. 53 (3), pp. 830-7. Date of Electronic Publication: 2015 Jan 07.
DOI:	10.1128/JCM.02648-14
Abstrakt:	A method for the rapid diagnosis of early dengue virus (DENV) infection is highly needed. Here, a prototype reverse transcription-recombinase 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 cross-reacting 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 IgG-capture 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 RT-LAMP 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. (Copyright © 2015, American Society for Microbiology. All Rights Reserved.)
Databáze:	MEDLINE
Externí odkaz:	Zobrazit plný text záznamu