Estabishment Of Rapid Diagnostic Methods For Detection Of Peste Des Petits Ruminants Virus,African Swine Fever Virus,Hedra Virus,Nipha Virus And West Nile Virus By Taqman Parallel Real-time PCR Assay

Due to peste des petits ruminants and African swine fever’s high infectious and high mortality caused the countries attach great importance to them.And Hedra disease,Nipha disease, West nile fever disease are zoonotic deadly infectious diseases that have been found in recent years. In July2007, PPR broke out in Ritu county, Ali area, Tibet of China. It was the first PPR outbreak in China. As an important transnational animal epidemic disease, PPR does a serious threat to the safety of animal heath and the development of stock raising in China. Therefore, enforcement actions of prevention, control and eradication must be adopted. African swine fever (ASF) is a highly lethal hemorrhagic disease of domestic swine, with mortality ratesapproaching100%. The causative agent, ASFV, is a unique and genetically complex DNA virus. ASF has been reported from most African countries south of the Sahara and more recently from Western and North African countries,European and American countries. Because no vaccine to prevent, these outbreaks were controlled by animal quarantine and slaughter, frequently at a very high cost. African swine fever has been reported in Russia. At this time, ASF is not present in China, but it is very important to prevent. Hedra virus (HeV)and Nipah virus (NiV) are newly recognized zoonotic viruses. HeV was discovered in Brisbane of Australia,1994,and NiV was discovered in Malaysia1999. It has caused severe diseases in animals and in humans through contact with infectious animals. HeV and NiV are named after the location where they were first detected. NiV is closely related to another newly recognized zoonotic virus, Hendra virus, named after the town where it first appeared in Australia in1994. Both Nipah and Hendraviruses are members of the virus family aramyxoviridae. West Nile virus is taxonomieally Placed within the family Flaviviridae, genus Flavivirus. Within the genus Flavivirus,WNV has been serologieally classified within the JEV antigenie complex,which includes the human Pathogens JE, Murray Valley encephalitis, SLE and Kunjin viruses. It is mosquito-bornevirus, Since its first isolating in1937in Uganda,it has become one of the most widespread flaviviruses worldwide. WNV can cause febrile illness,encephalitis and fatal meningoencephalitis in humans and birds.In addition to the transmission route of PPRV is direct contact with the spread or spread through droplets from a short distance, ASFV,HeV,NiV and WNV are spreaded by the middle of the media. The media of ASFV is ticks,HeV and NiV bats,WNV mosquito (Culex), these media are distributed and a wide variety in our country. Therefore, there is a larger risk that ASFV, HeV, NiV and WNV are irrupted into our country. It is much nessasry to establish a rapid, simple, specific and simultaneous diagnostic methods for preventing the viruses’ invading.In this study, according to the sequences of N gene of PPRV, P72gene of ASFV, H gene NiV, H gene NiV and PrM Gene of WNV reported in GenBank,we design twosets priers and probes for each type of virus,and then choose a better one with superior sensitivity and pecificity.45bat samples for HeV,NiV and WNV,38pig viscera samples for ASFV and20small ruminants secretions clinical samples for PPRV. The result of detection suggested that they were negative except the positive control. Estabishment of rapid diagnostic methods for detection of peste des petits ruminants virus,African swine fever virus,Hedra virus,Nipha virus and West Nile virus by Taqman parallel Real-time PCR assay has the specificity characteristics in this study, it can be used as the reference method.