大熊猫轮状病毒PCR检测方法的建立及应用

doi:10.16829/j.slxb.150454

兽类学报 ›› 2021, Vol. 41 ›› Issue (3): 254-260.DOI: 10.16829/j.slxb.150454

大熊猫轮状病毒PCR检测方法的建立及应用

苏小艳^1,2,3, 李林^1,2,3, 燕霞^1,2,3, 张东升^1,2,3, 侯蓉^1,2,3, 刘颂蕊^1,2,3

1 成都大熊猫繁育研究基地, 成都 610081;
2 四川省濒危野生动物保护生物学重点实验室, 成都 610081;
3 四川省大熊猫科学研究院, 成都 610081

收稿日期:2020-07-03 出版日期:2021-05-30 发布日期:2021-05-27
通讯作者: 侯蓉,E-mail:405536517@qq.com;刘颂蕊,E-mail:srui_liu@163.com
作者简介:苏小艳(1985-),助理研究员,主要从事大熊猫等野生动物疾病防控工作.
基金资助:
四川省科技计划项目（面上）（2018JY0363）；成都大熊猫繁育研究基地自立课题项目（2020CPB-B22）

Establishment and application of giant panda rotavirus PCR detection method

SU Xiaoyan^1,2,3, LI Lin^1,2,3, YAN Xia^1,2,3, ZHANG Dongsheng^1,2,3, HOU Rong^1,2,3, LIU Songrui^1,2,3

1 Chengdu Research Base of Giant Panda Breeding, Chengdu 610081, China;
2 Sichuan Key Laboratory of Conservation Biology for Endangered Wildlife, Chengdu 610081, China;
3 Sichuan Academy of Giant Panda, Chengdu 610081, China

Received:2020-07-03 Online:2021-05-30 Published:2021-05-27

摘要/Abstract

摘要： 轮状病毒是威胁大熊猫健康的主要病原微生物之一。为了对大熊猫轮状病毒进行快速、方便且准确地检测，研发适合于基层饲养单位和保护区的检测方法是非常有必要的。本研究通过合成大熊猫轮状病毒Vp7基因序列，构建了PUC-VP7的重组质粒，并将其作为阳性对照对大熊猫轮状病毒样本进行PCR检测和分析。结果表明，在进行PCR扩增分析时，该质粒和病毒cDNA二者均在340 bp处出现了特异性条带。此外，对收集到的45份大熊猫粪便样本进行轮状病毒抗原检测时，其中2份样品在340 bp处出现条带，该基因片段与大熊猫轮状病毒CH-1株的相似性为99.89%。本研究构建的PUC-VP7质粒不但可以作为大熊猫轮状病毒PCR检测中的阳性质控品，而且还能有效地促进该PCR病毒检测技术在基层饲养单位和保护区的推广和应用。

关键词: 大熊猫轮状病毒, PCR检测, Vp7基因, 质粒构建

Abstract: Rotavirus is one of the main pathogenic microorganisms threatening the survival of giant pandas. In order to detect panda rotavirus antigens quickly, conveniently and accurately, it is necessary to develop a detection method suitable for ex-situ breeding centers and protection areas. In this study, the panda rotavirus Vp7 gene sequence was synthesized to construct the PUC-VP7 recombinant plasmid, and it was used as a positive control for PCR detection and analysis of panda rotavirus samples. The results showed that during the PCR amplification analysis, both the plasmid and the viral cDNA showed a specific band at 340 bp. In addition, when performing rotavirus antigen detection on forty-five panda rotavirus fecal samples, two samples showed a band at 340 bp, and the gene fragment was 99.89% homologous to the panda rotavirus CH-1 strain. The PUC-VP7 plasmid constructed in this study can not only be used as a positive quality control material in the PCR detection of giant panda rotavirus, but can also effectively improve the promotion and application of the PCR virus detection technology in ex-situ breeding centers and protected areas.

Key words: Giant panda rotavirus, PCR test, Vp7 gene, Plasmid construction

中图分类号:

S858.92

苏小艳, 李林, 燕霞, 张东升, 侯蓉, 刘颂蕊. 大熊猫轮状病毒PCR检测方法的建立及应用[J]. 兽类学报, 2021, 41(3): 254-260.

SU Xiaoyan, LI Lin, YAN Xia, ZHANG Dongsheng, HOU Rong, LIU Songrui. Establishment and application of giant panda rotavirus PCR detection method[J]. ACTA THERIOLOGICA SINICA, 2021, 41(3): 254-260.

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大熊猫轮状病毒PCR检测方法的建立及应用

Establishment and application of giant panda rotavirus PCR detection method

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