Detection of bovine viral diarrhea virus by In situ hybridization

In situ hybridization에 의한 소 바이러스성 설사증 바이러스의 검출

Detection and distribution of bovine viral diarrhea virus(BVDV) was studied in formalin-fixed, paraffin-embedded tissues from two naturally infected cattle by in situ hybridization with a non-radioactive biotinylated probe. A 600 base pair cDNA probe from BVDV B-25 strain was used for probe. The whole procedure of ISH to diagnose was carried out within 1~2 hours in $Microprobe^{TM}$ capillary action system. The biotin-labelled probe was demonstrated after hybridization under standard conditions by the application of streptoavidin and biotinylated alkaline phosphatase. Alkaline phosphatase was visualized using a fast red TR/naphthol phosphatase and the sections were counterstained with hematoxylin. We have obtained the result of positive reactions in digestive tract(sm1.all intestine and colon) and epidermis of tongue in the state of the intact tissues. The result suggested that in situ hybridization method can be considered as a useful diagnostic technique for detection of specific nucleic acid sequences of BVDV.