Abstract
BackgroundMosquito-borne viruses (MBVs) and parasites (MBPs) are transmitted through hematophagous arthropods-mosquitoes to homoiothermous vertebrates. This study aims at developing a detection method to monitor the spread of mosquito-borne diseases to new areas and diagnose the infections caused by MBVs and MBPs.MethodsIn this assay, an ultra-sensitive chemiluminescence (CL) detection method was developed and used to simultaneously detect 19 common MBVs and MBPs. In vitro transcript RNA, virus-like particles (VLPs), and plasmids were established as positive or limit of detection (LOD) reference materials.ResultsMBVs and MBPs could be genotyped with high sensitivity and specificity. The cut-off values of probes were calculated. The absolute LODs of this strategy to detect serially diluted in vitro transcribed RNAs of MBVs and serially diluted plasmids of MBPs were 102–103copies/μl and 101–102copies/μl, respectively. Further, the LOD of detecting a strain of pre-quantified JEV was 101.8–100.8PFU/ml, fitted well in a linear regression model (coefficient of determination = 0.9678).ConclusionsUltra-sensitive CL imaging DNA hybridization was developed and could simultaneously detect various MBVs and MBPs. The method described here has the potential to provide considerable labor savings due to its ability to screen for 19 mosquito-borne pathogens simultaneously.
Highlights
Mosquito-borne viruses (MBVs) and parasites (MBPs) are transmitted through hematophagous arthropods-mosquitoes to homoiothermous vertebrates
Primers and probes design and evaluation In this assay, 12 MBVs and 7 Mosquito-borne parasites (MBPs) were chosen for detection: DENV1-4, Japanese encephalitis virus (JEV), yellow fever virus (YFV), West Nile virus (WNV), St
Complete genomic sequences of these MBVs, 18S rRNA gene sequences of malaria parasites, SspI repeat DNA sequence of W. bancrofti, and Hhal repeat region sequences of B. malayi and B. timori were downloaded from the GenBank database
Summary
Mosquito-borne viruses (MBVs) and parasites (MBPs) are transmitted through hematophagous arthropods-mosquitoes to homoiothermous vertebrates. This study aims at developing a detection method to monitor the spread of mosquito-borne diseases to new areas and diagnose the infections caused by MBVs and MBPs. Mosquito-borne viruses (MBVs) are transmitted from hematophagous arthropod/mosquitoes to homoiothermous vertebrates. Arthropods can support large virus inocula, and transmit them from an infected donor to a recipient during blood-feeding. Three major families in MBVs include the Togaviridae, Flaviviridae, and Bunyaviridae [1]. The globally pandemic MBV species encompass dengue virus (DENV), Japanese encephalitis virus (JEV), yellow fever virus (YFV), West Nile virus (WNV) [2] and chikungunya virus (CHIKV) [3]. In China, MBVs severely affect people living in tropical regions and rural areas.
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