Development Of The Techniques For The Detections Of Avian Influenza Virus And Comparison Of PB2 Gene Of The Guangxi Avian Influenza Isolates

Avian Influenza (AI) is an infection and/or disease syndrome caused by type A avian influenza virus (AIV). A variety of disease syndromes range from sub-clinical to mild upper respiratory disease to loss of egg production to acute generalized fatal disease. The highly pathogenic avian influenza (HPAI), caused by the H5N1 subtype has been causing seriously loss of economy in poultry industry and becoming a threat to the human health worldwide. Therefore, development of the technique to detect AIV quickly is the key for effective control the disease.A set of primers designed to amplify the matrix protein (MP) of AIV and another set of specific primers designed to amplify hemagglutinin (HA) protein of H5-subtype and a technique based on the reverse transcription-polymerase chain reaction (RT-PCR) was developed. The protocol for the detections of all the AIV isolates, the H5-subtype and the optimization of reactions were studied with a H5-subtype reference isolate BSG1. A 229bp-band of AIV specific and a 380bp-band of H5 subtype specific were amplified individually with the designed primers of AIV specific and H5-subtype specific respectively by the developed RT-PCR technique from isolate BSG1. A double-PCR was also developed which can detect all the AIVs and identify the H5-subtype AIVs. The results of specificity experiment showed that there was no specific band could be amplified with the samples of Newcastle disease virus, infectious bronchitis virus, infectious laryngotracheitis virus, egg drop syndrome virus and infectious bursal disease virus. The results of sensitivity experiment showed that the minimal amount of total sample...