Zhang Shu-Qin, Tan Bin, Li Peng, Wang Feng-Xue, Guo Li, Yang Yong, Sun Na, Zhu Hong-Wei, Wen Yong-Jun, Cheng Shi-Peng
State Key Laboratory for Molecular Biology of Special Economic Animals, Institute of Special Economic Animal and Plant Sciences, Chinese Academy of Agricultural Sciences CAAS, No. 4899 Juye Street, Chuangchun 130112, China.
State Key Laboratory for Molecular Biology of Special Economic Animals, Institute of Special Economic Animal and Plant Sciences, Chinese Academy of Agricultural Sciences CAAS, No. 4899 Juye Street, Chuangchun 130112, China.
J Virol Methods. 2014 Oct;207:204-9. doi: 10.1016/j.jviromet.2014.05.020. Epub 2014 Jul 12.
Bovine viral diarrhea virus (BVDV) can contaminate biological products produced in bovine or porcine cells or manufactured using bovine sera. A rapid, specific, sensitive, and practical method of detecting BVDV in bio-products is needed. The purpose of this study was to compare three assays with respect to their ability to accurately detect BVDV in biological samples, namely reverse-transcription loop-mediated isothermal amplification (RT-LAMP), SYBR green I-based real-time RT-PCR, and conventional RT-PCR. All assays detected BVDV nucleotide and differentiated between BVDV-free and -contaminated bovine sera successfully. In addition, the results were specific to BVDV: the amplification of samples containing the closely related classical swine fever virus or other pathogenic bovine viruses yielded negative results. The lowest detection threshold, 10(1) copies, was displayed by the SYBR green I-based real-time RT-PCR and RT-LAMP assay. This assay was also the most effective in the detection of BVDV contamination in a set of commercially available bovine sera. The field conditions suggest that RT-LAMP is specific and sensitive to detecting BVDV in biological samples and may be used for quality control of biomaterials.
牛病毒性腹泻病毒（BVDV）可污染在牛或猪细胞中生产或使用牛血清制造的生物制品。因此需要一种快速、特异、灵敏且实用的检测生物制品中BVDV的方法。本研究的目的是比较三种检测方法在准确检测生物样品中BVDV方面的能力，即逆转录环介导等温扩增（RT-LAMP）、基于SYBR Green I的实时RT-PCR和传统RT-PCR。所有检测方法均成功检测到BVDV核苷酸，并区分了不含BVDV和受BVDV污染的牛血清。此外，结果对BVDV具有特异性：含有密切相关的经典猪瘟病毒或其他致病性牛病毒的样品扩增结果为阴性。基于SYBR Green I的实时RT-PCR和RT-LAMP检测方法显示出最低检测阈值，为10(1)个拷贝。该检测方法在检测一组市售牛血清中的BVDV污染方面也是最有效的。现场情况表明，RT-LAMP对检测生物样品中的BVDV具有特异性和灵敏性，可用于生物材料的质量控制。
