The Establishment Of RT-PCR Detection Methods And Application To Reticuloendotheliosis Virus

In this study, we developed a RT-PCR assay to target the REV-LTR region, using the c DNA of one-day old REV infected chicken as a template. Based on the published sequence of REV-LTR, primers were designed and synthesized. Then a REV-PCR detection was established and optimized. On this basis, organs of chicken including heart, spleen, liver, lung, kidney, thymus and bursa of Fabricius were detected.Otherwise we detect fifty clinical blood samples from five farms of northeast China by biological detection technology and PCR method. The purpose of this study is to establish a rapid and efficient detection method of REV. To provide an important scientific basis for the prevention and treatment.The results were showen as follows:(1)The RT-PCR detection method to Reticuloendotheliosis virus have be successfully established.This method can effectively amplify the target fragment. The experience can be finished in 4 hours.(2)After optimized, the annealing temperature was fixed on 53℃, and the concentration of the primers and Mg2+ were fixed on 0.3μmol /L and 2.5μmol/L separately.(3)This method has a high sensitivity, specificity and repeatability.The results are highly consistent by testing these samples.This method has high repeatability and stability.(4)Using the above established RT-PCR detection method, the result shows that all the major organs of SPF chickens were infected, the positive rate of 50 clinical blood samples were up to 92%.