title: 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.
abstract: 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.
authors.affiliation: 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. Electronic address: shuqinzhang2008@163.com.
authors.forename: Shu-Qin
authors.initials: SQ
authors.lastname: Zhang
authors.affiliation: 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.
authors.forename: Bin
authors.initials: B
authors.lastname: Tan
authors.affiliation: 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.
authors.forename: Peng
authors.initials: P
authors.lastname: Li
authors.affiliation: 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.
authors.forename: Feng-Xue
authors.initials: FX
authors.lastname: Wang
authors.affiliation: 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.
authors.forename: Li
authors.initials: L
authors.lastname: Guo
authors.affiliation: 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.
authors.forename: Yong
authors.initials: Y
authors.lastname: Yang
authors.affiliation: 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.
authors.forename: Na
authors.initials: N
authors.lastname: Sun
authors.affiliation: 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.
authors.forename: Hong-Wei
authors.initials: HW
authors.lastname: Zhu
authors.affiliation: 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. Electronic address: yongjunwen@126.com.
authors.forename: Yong-Jun
authors.initials: YJ
authors.lastname: Wen
authors.affiliation: 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. Electronic address: tcscsp@126.com.
authors.forename: Shi-Peng
authors.initials: SP
authors.lastname: Cheng
mesh_terms: D000818:Animals; D052160:Benzothiazoles; D002417:Cattle; D006692:Classical Swine Fever Virus; D003959:Diamines; D008969:Molecular Sequence Data; D021141:Nucleic Acid Amplification Techniques / Q000379:methods*; D009930:Organic Chemicals / Q000378:metabolism; D010576:Pestivirus / Q000235:genetics / Q000302:isolation & purification*; D011804:Quinolines; D012367:RNA, Viral / Q000235:genetics / Q000302:isolation & purification*; D017422:Sequence Analysis, DNA; D044967:Serum / Q000821:virology*; D013194:Staining and Labeling / Q000379:methods; D013696:Temperature; D013997:Time Factors
chemical_list: D052160:Benzothiazoles; D003959:Diamines; D009930:Organic Chemicals; D011804:Quinolines; D012367:RNA, Viral; C098022:SYBR Green I
keywords: Bovine viral diarrhea virus; RT-PCR; Reverse-transcription loop-mediated isothermal amplification; SYBR green I-based real-time RT-PCR