The Design Of AIV H5,H7,H9 RT-PCR Detection Kit

Avian influenza virus,called the AIV,is an acute respiratory tract infectious disease which caused by a type A influenza virus.AIV granule surface's haemagglutinin and neuraminic acid enzyme have the genotype characteristic.After AIV H5N1 infected the humanity in 1997,H9N2,H7N7 genotype infected the humanity and H5N1 one after another infected humanity had been reported once more,this has aroused people's widespread interest.When you understand the entire gene order,or only understand the fragment both sides(5 'ends,3 'ends) sequence,you can increases goal gene DNA fastly in the way PCR.PCR is highly effective, specially,is faithful highly to the template,and its production is highly pure,the production can be used in researching work directly.From PCR had been inventioned in the 1990s to now,it is still one kind of important method of increase DNA in vitro.Therefore,diagnosis AIV and its genotype through the PCR technology is one ideal methods of prevention AIV.This experiment withdraws and reverses transcripe RNA from AIV H5,H7,H9,and increases them.The result indicated that every production of PCR is right with the primers(the fragments size of H5,H7,H9 are 420bp,228bp,830bp).In the foundation of H5,H9 multiplex PCR and H7,H9 multiplex PCR,the experiment establishes H5,H7,H9 multiplex PCR.Then,the experiment find the best mix system of PCR under the different condition.This RT-PCR Detection Kit can diagnosis AIV in 1～2h.This production is advantageous to the scientific research or the prevention of AIV.The result indicated that when the PCR system has buffer 2.5μl/25μl,dDNTP 2μl/25μl,rtaqE 0.25μl/25μl,H5 primer 0.2μl/25μl,H7 primer 0.4μl/25μl,H9 primer 0.6μl/25μl,this system of multiplex PCR is the best,this has been proved by the specificity and the sensitivity experiment.Then after long time under different PCR premix system and preserved condition test,the experiment finds the final PCR mix system is Buffer+ water +dDNTP+rtaqE+primers,this mix system can be preserved for a long time under - 20℃,and designs actual production of RT-PCR Detection Kit finally.