Detection of bovine viral diarrhea virus by In situ hybridization

Park, Nam-yong; Hong, Ki-kang; Chung, Ci-young; Cho, Kyoung-oh; Lee, Bong-joo; Park, Young-seok; Park, Hyung-seon; Kweon, Chang-hee

Korean Journal of Veterinary Research 1999;39(1):138-147.

Nam-yong Park¹, Ki-kang Hong¹, Ci-young Chung¹, Kyoung-oh Cho¹, Bong-joo Lee¹, Young-seok Park¹, Hyung-seon Park¹, Chang-hee Kweon²

¹College of Veterinary Medicine, Chonnam National University
²National Veterinary Research and Quarantine Service

In situ hybridization에 의한 소 바이러스성 설사증 바이러스의 검출

박남용¹, 홍기강¹, 정치영¹, 조경오¹, 이봉주¹, 박영석¹, 박형선¹, 권창희²

¹전남대학교 수의과대학
²국립수의과학검역원

Abstract

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.

Key Words: bovine viral diarrhea, in situ hybridization, cattle, Togavirus, BVD