The Identification Of Bovine Rotavirus And Rapid Detection Method Study

The identification of a rotavirus and the cloning of the VP7 gene Rotavirus is a main pathogeny that can cause serious diarrheas. If the calf caught the rotavirus disease, the speed of the grove can be slowed because of the function disorder of the intestins .Meanwhile the consistent diarrhea can arise the economy dehydration, even the death of the calves. It is 1982 that the rotavirus was identified for the first time in buffalo, therefore the gallinaceous rotavirus, swine and rabbit rotavirus was identified in different districts. The rotavirus that arises the serious diarrhea in calves are mainly the G6 serotype. The American professor ANITL V PARWANT adopted the method of nucleic acid probe to detect different serotypes in the cattle,and the result suggests that the G6 serotype are the main serotype in American (G6,36.8%;G8,2.9%;G10,12.7%) .But there are few of the reference about the rotavirus in China. The study mostly focuses on the identification of the rotavirus, and the pathology and the rapid detection method are rarely studied.Clinical specimens Feces and duodenum were collected from a calf herd in ShanDong province in which broke out a severe diarrhea, and the biology characteristic was studied. Virus isolation and propagation MA-104 cells (supported by China center for type culture collection) were cultured on routine rules with Dulbecco's Modified Eagles Medium. The isolate was well adapted to the MA-104 and the cell pathological changes appeared when the sample was serial cultured for four times.The rotavirus has adapted to the 27 times, the cell pathology was appeared in the 28-36 hours. The study of electro microscope pathology in the MA-104 cells suggests that the rotavirus can increase in the MA-104 cells, the swelling of the matrix, and the membrane of the cell became cracking. But in the experiment, we found that the number of the rotavirus that can successively affect the health cells is lower than the total number, there may have the relation with the specific cultivation of the rotavirus. The VP7 gene of the isolates was 1062 base pairs in length and contained an open reading frame of 981 base pairs that encoded a 323 amino acid. The nucleotide sequence of the VP7 genes is of high identity. The primers were designed compared with the NCDV and IND strain's VP7 gene in the methods of the DNA Star. And the method of RT-PCR was adopted in order to extent the segment of...