Journal Article

An RT-PCR panel for rapid serotyping of dengue virus serotypes 1 to 4 in human serum and mosquito on a field-deployable PCR system

Lee, Fu-Chun, Ping, Chia-Fong, Tsai, Jih-Jin, Chen, Chun-Hong, Lin, Ping-Chang, Tsai, Ching-Yi, Chou, Pin-Hsing, Lee, Pei-Yu Alison, Liu, Li-Teh, Liu, Wei-Liang, Huang, Bo-Yi

2019

Overview

Dengue fever, a mosquito-borne disease, is caused by dengue virus (DENV) which includes four major serotypes (DENV-1, -2, -3, and -4). Some serotypes cause more severe diseases than the other; severe dengue is associated with secondary infections by a different serotype. Timely serotyping can provide early warning of dengue epidemics to improve management of patients and outbreaks. A mobile insulated isothermal PCR (iiPCR) system is available to allow molecular detection of pathogens near points of need. In this study, side-by-side comparison with the CDC DENV-1-4 Real Time RT-PCR (qRT-PCR) was performed to evaluate the performance of four singleplex DENV-1-4 serotyping reverse transcription-iiPCR (RT-iiPCR) reagents for DENV subtyping on the mobile PCR system. The four RT-iiPCRs did not react with Zika virus and chikungunya virus; tests with serial dilutions of the four DENV serotypes made in human serum showed they had detection endpoints comparable to those of the reference method, indicating great analytical sensitivity and specificity. Clinical performance of the RT-iiPCR reagents was evaluated by testing 40 serum samples each (around 20 target serotype-positive and 20 DENV-negative); all four reagents had high agreement (97.5-100%) with the reference qRT-PCR. Moreover, testing of mosquitoes separately infected experimentally with each serotype showed that the four reagents detected specifically their target DENV serotypes in mosquito. With analytical and clinical performance comparable to the reference qRT-PCR assay, the four index RT-iiPCR reagents on the field-deployable PCR system can serve as a useful tool for DENV detection near points of needs.

• Publisher: Public Library of Science,Public Library of Science (PLoS)
• Subject: Animals / Antigens / Aquatic insects / Biology and life sciences / Biotechnology / Chikungunya virus / Culicidae - virology / Dengue / Dengue - virology / Dengue fever / Dengue hemorrhagic fever / Dengue virus / Dengue Virus - classification / Dengue Virus - genetics / Disease control / Epidemics / Female / Fever / Health aspects / Health surveillance / Hospitals / Humans / Hygiene / Infection / Infections / Infectious diseases / Insect Vectors - virology / Laboratories / Medical research / Medical technology / Medicine / Medicine and Health Sciences / Methods / Mosquitoes / Outbreaks / Pathogenic microorganisms / Point-of-Care Systems / Polymerase chain reaction / Public health / Reagents / Research and Analysis Methods / Reverse Transcriptase Polymerase Chain Reaction / Reverse transcription / RNA, Viral - blood / Sensitivity analysis / Sensitivity and Specificity / Serotypes / Serotyping / Target detection / Underserved populations / Vector-borne diseases / Viral diseases / Viruses / Zika virus