Epidemiological Surveillance And Molecular Analysis For Three Neurotropic Virus Infection In Western Region Of China

ObjectiveTo investigate the natural infection and epidemic of Nipah virus (NiV) and Hendra virus (HeV) in animal in western region of China, obtain the epidemiologic data of NiV and HeV in western region of China and assess of the biosafety status in these areas. To detect the Hepatitis C Virus (HCV) RNA strands of unknown pathogen viral encephalitis patients of Chongqing region, investigate the relationship between HCV and human viral encephalitis, and find the role of HCV infection in human nerve and mental disorders.Methods1.Randomly collect brain tissues of 183 grazing horses and 65 donkeys, free vaccination, in in Ili and Bain, Xinjiang area. Grind brain tissues and extract total RNA with Trizol method. Detect NiV of the RNA samples with one-step real-Time RT-PCR, and submit epidemiological investigation.2.Randomly collect brain tissues of 183 grazing horses and 65 donkeys, free vaccination, in in Ili and Bain, Xinjiang area. Grind brain tissues and extract total RNA with Trizol method. Detect HeV of the RNA samples with one-step real-Time RT-PCR, and submit epidemiological investigation.3.Randomly collect brain tissues of 183 grazing horses and 65 donkeys, free vaccination, in in Ili and Bain, Xinjiang area. Grind brain tissues and extract total RNA with Trizol method. Detect HeV of the RNA samples with one-step real-Time RT-PCR, and submit epidemiological investigation.Results1.The detectable concentration of NiV reached to 1.1×102-1.1×107copies/μl by one-step real-Time RT-PCR method. The HeV N gene and BDV nucleoprotein gene fragment was negative of all samples.2.The NiV N gene fragment of horse and donkey brain samples from Ili area was detected by one-step real-Time RT-PCR method successfully. There was no Takeoff point on fluorescence amplification curves of all samples. Curves kept the same slope, and assays were judged as negative.3.The detectable concentration of HeV reached to 2.6×102-2.×107copies/μl by one-step real-Time-RT-PCR method. And the detection of NiV N gene and BDV nucleoprotein gene fragment was negative of all samples.4.The HeV N gene fragment of horse and donkey brain samples from Ili area were detected by one-step real-Time RT-PCR method successfully. There was no Takeoff point on fluorescence amplification curves of all samples. Curves kept the same slope, and assays were judged as negative.5.The detectable concentration of HCV positive and negative-strand RNA 5'NCR gene fragment reached to 7.85×100-7.85×107copies/μl by nest real-Time RT-PCR method. Breakbone fever virus 5'NCR gene, NiV N gene and WNV E gene fragment were negative by the method.6.Peripheral blood samples and cerebrospinal fluid samples of 64 patients with viral encephalitis and were collected from Chongqing. HCV positive and negative-strand RNA 5'NCR gene fragment of all samples were detected by one-step real-Time RT-PCR method successfully. One positive cases (1/32) of 5'NCR positive-strand fragment was detected in nucleated cells in cerebrospinal fluid and two (2/32) in PBMC while one positive case (1/32) and none (0/32) in negative-strand fragment in the 32 cases.Conclusions1.The Nipah virus one-step real-time RT-PCR is sensitive and reliable and allows rapid detection and quantitation of NiV in field and experimental materials, and is useful for epidemiological surveillance and specific diagnosis.2.There was no infection of NiV in either horses or donkeys brain tissue samples from Ili, Xijiang area.3.The Hendra virus one-step real-time RT-PCR is sensitive and reliable and allows rapid detection and quantitation of HeV in field and experimental materials used for epidemiological surveillance and specific diagnosis.4.There was no infection of HeV in either horses or donkeys brain tissue samples from Ili, Xijiang area.5.The HCV nest fluorescent quantitation real-time RT-PCR is sensitive and reliable and allows rapid detection and quantitation of NiV in field and experimental materials, and is useful for epidemiological surveillance, specific diagnosis and the research of the relationship of HCV infection and nerve disease.6.There were HCV infection samples from peripheral blood and cerebrospinal fluid samples of viral encephalitis patients collected from Chongqing, which implied the correlation between HCV infection and viral encephalitis.