Investigation Of Arboviruses In Liaoning And Heilongjiang Provinces

Arboviruses can be defined as the set of virus transmitted by blood-sucking hexapod, which can cause zoonose. There are only four kinds of arboviruses currently present and prevalent in China. But there must be more than 4 kinds of arboviruses in China. So it's vital to isolate and identify arboviruses to provide theoretical foundation to establish new step for diseases control and prevention.In this study, more than 7900 mosquitoes were collected in Liaoning and Heilongjiang provinces in 2002. The homogenates of mosquitoes were inoculated onto BHK-21 cells lines. Four strains were derived, named LN02-102, LN02-104, HLJ02-134, HLJ02-136, respectively. They caused cytopathogenic effect (CPE) both in BHK-21 and C6/36 cells and were fatal to sucking mice.Firstly, the newly isolated strains were identified by serological methods. Both Enzyme-linked Immunosorbent assay(ELISA) and Immunofluoresence assay(IFA) showed that all the 4 strains were positive to the standard antibody of JE virus.Secondly, Real-Time PCR was used to identify the viruses. The amplification curves showed that the 4 strains were JEV. And the PrM and E gene of the newly isolated JEV strains were amplified by Reverse Transcript Polymerase Chain Reaction(RT-PCR) and sequenced.Finally, to analysis the genotype, 240 nucleotides of PrM region(map position 456-695) of the new isolats were selected and compared with the reference strains from GenBank, which were isolated from different countries, and sources. The result showed that LN02-102, LN02-104 isolated in Liaoning belonged to genotype I , JEV, which is the first report in Liaoning province. And HLJ02-134, HLJ02-136 isolated in Heilongjiang belonged to genotype III,...