Comparison of conventional RT-PCR, reverse-transcription loop-mediated isothermal amplification, and SYBR green I-based real-time RT-PCR in the rapid detection of bovine viral diarrhea virus nucleotide in contaminated commercial bovine sera batches
文献类型: 外文期刊
第一作者: Zhang, Shu-Qin
作者: 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
作者机构:
关键词: Bovine viral diarrhea virus;RT-PCR;Reverse-transcription loop-mediated isothermal amplification;SYBR green l-based real-time RT-PCR
期刊名称:JOURNAL OF VIROLOGICAL METHODS ( 影响因子:2.014; 五年影响因子:2.001 )
ISSN:
年卷期:
页码:
收录情况: SCI
摘要: 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, 101 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
分类号: R37
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