猪圆环病毒II型PCR检测方法的研究

摘要/Abstract

摘要： 【目的】建立快速、简便、特异的检测猪圆环病毒II型的PCR方法；【方法】根据已发表的猪圆环病毒II型基因序列设计合成了一对引物，用酚-氯仿法抽提可疑病料中的病毒DNA，应用PCR技术对猪圆环病毒进行基因扩增，PCR产物进行序列测定；以猪瘟病毒、猪伪狂犬病毒、猪细小病毒为对照，进行特异性试验；取部分病料进行重复性试验；【结果】PCR方法扩增出了长度为702bp的片段，扩增产物经测序和序列分析表明扩增的序列为PCV2序列；猪瘟病毒、猪伪狂犬病毒、猪细小病毒的PCR扩增均为阴性,与预期结果一致；部分病料PCR重复检测5次，检测结果完全一致；【结论】PCR检测方法可以对猪圆环病毒病作出敏感、特异、准确地诊断，该方法检测的基因最低模板量为2×10-5 ng/ml

关键词: 柑桔黄龙病, 柑桔黄龙病, 单克隆抗体, 胶体金, 快速检测

Abstract: 【Objective】A polymerase chain reaction for detecting PCV2 was established；【Mothed】Based on the gene sequence of PCV2, a pair of primers was designed.viral DNA from the sick tissues suspious of inclouding PCV2 extracted with phenol/chloroform as template, PCR technique was used; the special test of PCR detection was taken with the Hog choleravirus,Pseudorabies virus and Swine Parvovirus virus as control;the repetitive test of PCR detection was also taken.【Result】A size of 702 bp was amplified by PCR，and the PCR production was sequenced, sequence analysis showed that the sequence belonged to the genome of PCV2; the Hog choleravirus,Swine Pseudorabies virus and Swine Parvovirus virus were also detected by the PCR as control under the same condation with negative results; some sick tissues were detected for 5 times with high repetitive results.【Conclusion】The method of PCR detection was a sensitive, specific and reliable method for detecting PCV2, the sensitivity of PCR detection for PCV2 was up to 2×10-5ng／ml DNA of positive cell virus infected PCV2.

中图分类号:

S852.659

王宪文,刘兴友,梁美兰,郑玉姝,刘丽艳. 猪圆环病毒II型PCR检测方法的研究[J]. 中国农学通报, 2007, 23(9): 8-8.

Wang Xianwen, Liu Xingyou, Liang Meilan, Zheng Yushu, Liu Liyan. Detection of Procine Circovirus Type 2 by PCR Technique[J]. Chinese Agricultural Science Bulletin, 2007, 23(9): 8-8.

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