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首页> 外文期刊>Journal of Virological Methods >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
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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

机译：常规RT-PCR，逆转录环介导的等温扩增和基于SYBR Green I的实时RT-PCR在快速检测受污染的商业牛血清批次中的牛病毒性腹泻病毒核苷酸中的比较

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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
机译：牛病毒性腹泻病毒（BVDV）会污染牛或猪细胞中产生的或使用牛血清制造的生物产品。需要一种快速，特异性，灵敏和实用的方法来检测生物产品中的BVDV。这项研究的目的是比较三种测定法在准确检测生物样品中BVDV的能力方面，即逆转录环介导的等温扩增（RT-LAMP），基于SYBR green I的实时RT-PCR，和传统的RT-PCR。所有测定均检测到BVDV核苷酸，并成功区分了无BVDV和受污染的牛血清。此外，该结果是特定于BVDV的：扩增包含密切相关的经典猪瘟病毒或其他病原性牛病毒的样品产生阴性结果。最低检测阈值为101份，通过基于SYBR green I的实时RT-PCR和RT-LAMP分析法显示。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。现场条件表明，RT-LAMP对检测生物样品中的BVDV具有特异性和敏感性，可用于生物材料的质量控制。

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