利用IC-RT-PCR检测怀地黄中烟草花叶病毒（TMV）
[Detection of tobacco mosaic virus (TMV) in Rehmannia glutinosa f. hueichingensis by IC-RT-PCR].
作者信息
Du Lin, Xiang Jin-Le, Fan Jin-Ling, Li Xin, Luo Lei
机构信息
Food and Bioengineering College, Henan University of Science and Technology, Luoyang 471023, China.
出版信息
Zhongguo Zhong Yao Za Zhi. 2013 Jul;38(13):2092-5.
OBJECTIVE
To establish a rapid, sensitive and efficient detection method for tobacco mosaic virus (TMV), and provide technical support of TMV detection of Rehmannia glutinosa f. hueichingensis. The virus-free plantlets could be produced on a large scale to ameliorate breed degeneration caused by viral disease.
METHOD
Specific primers were designed based on the conserved region of coat protein(CP) gene of TMV. Immunocapture RT-PCR (IC-RT-PCR) was employed to detect TMV and the sequence of the products was detected.
RESULT
The expected nucleotide acid fragments were amplified by IC-RT-PCR. The homology of nucleotide acid sequence and amino acid sequence were 95.29% and 96.7% between the PCR products and the CP gene of TMV (accession number AY555269).
CONCLUSION
The method was established for the detection of TMV in R. glutinosa f. hueichingensis by IC-RT-PCR. This detection combined molecular biology technology with immunology, was convenient for a quick, sensitive and simple detection of TMV.
目的
建立一种快速、灵敏、高效的烟草花叶病毒（TMV）检测方法，为怀地黄TMV检测提供技术支持，以便大规模生产无病毒苗，改善病毒病引起的品种退化。
方法
根据TMV外壳蛋白（CP）基因保守区设计特异性引物，采用免疫捕捉RT-PCR（IC-RT-PCR）检测TMV，并对产物进行测序。
结果
IC-RT-PCR扩增出预期核苷酸片段，PCR产物与TMV的CP基因（登录号AY555269）核苷酸序列同源性为95.29%，氨基酸序列同源性为96.7%。
结论
建立了IC-RT-PCR检测怀地黄中TMV的方法。该检测方法将分子生物学技术与免疫学相结合，便于快速、灵敏、简便地检测TMV。
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