Study On Mid-East Interactive Development And Economic Cooperation In Mid-China’s Rising

Objective:According to the Japanese encephalitis virus sequenced by Genebank and NCBI specific Blast primers and Oligo6.0 software.By constantly optimized the PCR assay conditions, using different reaction temperature,time and primer concentrations to determine the optimal testing conditions, to establish the method of detecting Japanese encephalitis virus by RT-PCR.Methods:The research included 21 epidemic encephalitis B patients as the experimental group, 20 cases of healthy people were collected as control group.The cases were collected during June to October in 2013 in our hospital. Polymerase Chain Reaction(PCR) and enzyme linked immunosorbent(ELISA) were used to detect blood samples from patients and control group.First, Japanese encephalitis virus were isolated from blood samples, and inoculate the virus to BHK-21 hamster kidney cells.The virus were collected after the propagation culture. According to the Japanese encephalitis virus sequenced by Genebank and NCBI specific Blast primers and Oligo6.0 software.By constantly optimized the PCR assay conditions, using different reaction temperature,time and primer concentrations to determine the optimal testing conditions, to establish the method of detecting Japanese encephalitis virus by RT-PCR. Reverse Transcription-Polymerase(RT-PCR) method were used to test the serum of 87 cases of suspected Japanese encephalitis patients in our hospital between June 2013 and December 2014. Detecting the accuracy and sensitivity between RT-PCR and ELISA.Results:We got the best effect when the PCR conditions were 50℃, 60 min, primer 1μl PCR reaction when added. The results showed that PCR positive test results was95.2%, PCR products were sequenced and turned to be the accuracy rate of that was100%. Clinical application of evaluation showed PCR detection accuracy can reach96.8%.Conclusion:RT-PCR has a high diagnostic accuracy, it is possible to shorten the time of diagnosis, reduce diagnostic costs.It can also minimize the harm to patients, the diction method could be extended in terms of clinical diagnosis encephalitis.