The Design Of One Step Fluorescent RT-PCR Kit For Simultaneous Detection Of The AI-HPAI-ND Virus

Avian influenza and Newcastle disease are both the most important avian viralzoonosis in the World. These two viral diseases have significant impacts on thepoultry industry. In recent years, the incidence and prevalence of avian influenza andnewcastle disease not only seriously affected the consumer confidence, but alsohindered the international poultry products trade. It is the most puzzling problem inpoultry industry nowadays that the subtypes of influenza H5, H7, H9enhance theharm and increase directly infecting number. The conventional technology detectingavian influenza and newcastle disease, such as agar gel diffusion （AGP）,hemagglutination inhibition test （HI）, the embryonated egg virus isolation test andELISA method are difficult to make a quick diagnose, so it needs to establish a rapid,accurate and efficient detecting method.Real-time quantitative RT-PCR （real-time fluorescent quantitative RT-PCRtechnology） technology is a new molecular nucleic acid detecting technology. Wedesigned specific primers and probes according to conserved sequence of avianinfluenza virus gene M, gene HA, newcastle disease virus gene M and establish asensitive, specific and rapid one-step fluorescence quantitative RT-PCR detection ofgeneral type, subtype H5, H7, H9of avian influenza virus and newcastle disease virus.Then we will develop a quick diagnostic kit that could detect samples in a short timeand determine the presence of avian influenza virus and newcastle disease viruseventually. With the people’s demand for green products as well as the exportinspection and quarantine standards increasing, this technology will have a broadmarket prospect.It is inevitable to be a waste of labor force, materials and time if we take onlyone probe and primer accordance with certain subtype of avian influenza virus andtest by subtype in the inspection and quarantine of the practical work. Therefore, itwill greatly save the detecting time using general probe and primer to verify avian fluvirus first. Then use probes and primers to subtype H5, H7and H9if avian influenzavirus exists, A virus probe, primer genotyping. Universal fluorescent RT-PCR can detect the avian influenza virus infection with allantoic fluid highest dilution up to10^-10as well as other subtypes fluorescent RT-PCR...