Direct polymerase chain reaction from blood and tissue samples for rapiddiagnosis of bovine leukemia virus infection

Journal of Veterinary Medical Science

Kazuhiko OHASHI¹  Satoru KONNAI¹  Ayako NAKAHARA¹  Asami NISHIMORI¹  Shiro MURATA¹  Tomohiro OKAGAWA¹  Ryoyo IKEBUCHI¹ 

[1] Department of Disease Control, Graduate School of Veterinary Medicine, Hokkaido University, Kita-ku Kita 18- jo Nishi 9-chome, Sapporo, Hokkaido 060�?0818, Japan

关键词: bovine leukemia virus (BLV);    PCR directly from whole blood;    provirus;   

DOI  :  10.1292/jvms.15-0577

学科分类：兽医学

来源: Japanese Society of Veterinary Science

【 摘 要 】

References(17)Bovine leukemia virus (BLV) infection induces bovine leukemia in cattle and causessignificant financial harm to farmers and farm management. There is no effective therapyor vaccine; thus, the diagnosis and elimination of BLV-infected cattle are the mosteffective method to eradicate the infection. Clinical veterinarians need a simpler andmore rapid method of diagnosing infection, because both nested polymerase chain reaction(PCR) and real-time PCR are labor intensive, time-consuming, and require specializedmolecular biology techniques and expensive equipment. In this study, we describe a novelPCR method for amplifying the BLV provirus from whole blood, thus eliminating the need forDNA extraction. Although the sensitivity of PCR directly from whole blood (PCR-DB) samplesas measured in bovine blood containing BLV-infected cell lines was lower than that ofnested PCR, the PCR-DB technique showed high specificity and reproducibility. Among 225clinical samples, 49 samples were positive by nested PCR, and 37 samples were positive byPCR-DB. There were no false positive samples; thus, PCR-DB sensitivity and specificitywere 75.51% and 100%, respectively. However, the provirus loads of the samples detected bynested PCR and not PCR-DB were quite low. Moreover, PCR-DB also stably amplified the BLVprovirus from tumor tissue samples. PCR-DB method exhibited good reproducibility andexcellent specificity and is suitable for screening of thousands of cattle, thus servingas a viable alternative to nested PCR and real-time PCR.

【 授权许可】

Unknown