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
Jih-Jin Tsai,, Wei-Liang Liu, Ping-Chang Lin, Bo-Yi Huang, Ching-Yi Tsai, Pin-Hsing Chou, Fu-Chun Lee, Chia-Fong Ping, Pei-Yu Alison Lee, Li-Teh Liu, Chun- Hong Chen| Center for Dengue Fever Control and Research, Kaohsiung Medical University, Kaohsiung, Taiwan, School of Medicine, College of Medicine, Kaohsiung Medical University, Kaohsiung, Taiwan; Tropical Medicine Center, Kaohsiung Medical University Hospital, Kaohsiung, Taiwan; Division of Infectious Diseases, Department of Internal Medicine, Kaohsiung Medical University Hospital, Kaohsiung, Taiwan; National Mosquito-Borne Diseases Control Research Center, National Health Research Institutes, Zhunan, Taiwan; GeneReach Biotechnology, Taichung, Taiwan; Department of Medical Laboratory Science and Biotechnology, College of Medical Technology, Chung-Hwa University of Medical Technology, Tainan City, Taiwan; National Institute of Infectious Diseases and Vaccinology, National Health Research Institutes, Zhunan, Taiwan|PLoS ONE|2019|14, 3, e0214328, https://doi.org/10.1371/journal. pone.0214328
Dengue fever is a mosquito-borne disease that is caused by dengue virus. Dengue disease is a major public health concern in developing tropical countries, with frequent international travel causing escalating numbers of imported dengue cases to new geographical regions. The virus has four major serotypes (DENV-1, -2, -3 and -4), with some causing more severe diseases than the others. Severe dengue is associated with secondary infection by a different serotype. Clinical diagnosis cannot be relied on as majority of infections are either asymptomatic or present with symptoms similar to those of other febrile-episode-inducing diseases. Therefore, timely on-site detection and serotyping of dengue virus in humans and mosquitos can allow for timely intervention and help mitigate disease outbreaks in humans. The current rapid commercially available diagnostic tests do not provide serotype information and if they can they require highly skilled technicians and expensive equipment that cannot be accessed in remote areas. In order to bring early serotyping of dengue virus to points of need, a rapid, easy, mobile PCR method of high sensitivity and specificity is needed. A side-by-side comparison with the CDC DENV-1-4 Real Time RT-PCR (qRT- PCR) on the Mic qPCR cycler 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 POCKIT system. All data indicated that the four DENV-1, -2, -3, and -4 RT-iiPCR had analytical sensitivity comparable to that of the reference qRT-PCR in detecting their target DENV serotypes.