三种不同引物扩增对猪圆环病毒2型检测敏感性的研究
毕业论文关键词：猪圆环病毒2型；不同引物；PCR；敏感性
Comparative study on the sensitivity of three kinds of different primers for the detection of Porcine circovirus type 2
Abstract：In order to detect exogenous porcine circovirus type 2 (PCV2) in vaccine products rapidly, we designed a pair of primers Y1 and Y2 on the basis of the ORF2 gene for PCR detection,and compared its specificity, sensitivity with PCV national standard primers (P1/P2/P3) and entry-exit inspection primer. The results showed that (1) Three kinds of primers had high specificity for the detection of PCV2; (2) When the DBN-SX07 strain was detected , the PCRs using primers Y1/Y2 and P1/P2/P3 were able to detect at least 0.079TCID50 PCV2, while the nest-PCR primers nP1/nP2/nP3/nP4 could detect only 7.9 TCID50 ; (3) When the clinically acquired strain was been detected, the PCR using primers Y1/Y2 was able to detect at least 1.58×10-6TCID50 PCV2, while the primers P1/P2/P3 and nP1/nP2/nP3/nP4 could detect only 1.58×10-4TCID50. (4) When the virous titer is high, the results of the nest-PCR primers nP1/nP2/nP3/nP4 are always confused. Consequently, the primers Y1/Y2 had higher sensitivity for the detection of PCV2, and it was more suitable for the detection of exogenous PCV2 in vaccine products.
Key words: PCV2; Different primers; PCR; specificity
目 录
摘要1
关键词1
Abstract1
Key words1
引言1
1材料与方法2
1.1材料 2
1.1.1主要仪器 2
1.1.2 细胞和病毒 2
1.1.3酶和试剂 2
1.1.4引物 2
1.2方法 3
1.2.1 供试病毒TCID50的测定3
1.2.2 病毒DNA的提取 3
1.2.3 PCR检测3
1.2.4琼脂糖凝胶电泳5
1.2.5特异性和敏感性分析5
1.2.6重复性分析5
2 结果5
2.1毒株的TCID505
2.2特异性分析6
2.3敏感性分析7
2.4重复性分析8
3 讨论 8
致谢8
参考文献9
附录10
三种不同引物扩增对猪圆环病毒2型检测敏感性的研究
猪圆环病毒（PCV）是目前世界上体积最小、无囊膜的单链DNA病毒，现已知猪圆环病毒有两个基因型：PCV1和PCV2，其中PCV1没有致病性，只有PCV2具有致病性。PCV2现在被认为是仔猪断奶后多系统衰竭综合征（PMWS）的主要病原，此外，它还与母猪流产、仔猪先天性震颤（CT）、育肥猪的慢性呼吸道病综合征（PRDC）和猪皮炎和肾病综合症（PDNS）等疾病有关[1]。因此，PCV2感染会给养猪业造成严重的经济损失。
在疫苗生产中，由于所用原材料均来源于动物，存在外源病毒污染的风险。猪圆环病毒是疫苗制备中可能出现的外源病毒之一，2010年便出现过一次外源PCV污染轮状病毒疫苗事件[2]，影响较大。因此，疫苗尤其是减毒活疫苗的生产中对PCV2的检测极为重要。检测外源病毒时，最常用的检测手段是PCR，现在我国已将检测猪圆环病毒的PCR方法列为国家标准。
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