4种猪群常见病毒基因芯片检测方法的建立与应用  

Development of a genechip for detection of CSFV,PRRSV,PCV-2 and PPV

作　　者：杨若松[1,2] 姜金庆[1] 张志[2] 李晓成[2] 王建华[1] 任夫波[3] 张丽丽[3] 

机构地区：[1]西北农林科技大学动物医学院,陕西杨凌712100 [2]中国动物卫生与流行病学中心,山东青岛266032 [3]青岛农业大学动物科技学院,山东青岛266032

出　　处：《西北农林科技大学学报（自然科学版）》2012年第3期34-38,共5页Journal of Northwest A&F University(Natural Science Edition)

基　　金：国家"十一五"科技支撑计划项目(2007BAD86B05)

摘　　要：【目的】建立猪瘟病毒(CSFV)、高致病性猪蓝耳病病毒(PRRSV)、猪圆环病毒2型(PCV-2)及猪细小病毒(PPV)的基因芯片检测方法,为这4种病的实验室诊断和流行病学调查提供一种快速、高效的病原学诊断方法。【方法】根据GenBank中CSFV、高致病性PRRSV、PCV-2、PPV的相关基因序列,设计合成相关引物,采用PCR方法得到具有荧光标记的PCR产物,然后与含有特定检测探针的芯片进行杂交,建立其基因芯片检测方法,并对该方法的灵敏性和重复性进行检测。应用建立的基因芯片检测方法,对150份临床样品进行检测。【结果】建立了针对CSFV、PRRSV、PCV-2和PPV 4种病毒的基因芯片检测方法,该方法对临床150份样品的检测结果显示,CSFV阳性样品有25份,高致病性PRRSV阳性样品有45份,PCV-2阳性样品有70份,PPV的检测结果全部为阴性,其中混合感染样品有18份。【结论】建立了对CSFV、PRRSV、PCV-2、PPV 4种猪群常见病毒的基因芯片检测方法,该方法具有良好的特异性、敏感性和重复性。[Objective] The detection method of CSFV,PRRSV,PCV-2 and PPV is designed by Gene- chip technique,which is a fast, highly active method for lab diagnosis and epidemiological investigation of the four diseases. [Method] According to the sequence of CSFV,PRRSV,PCV-2 and PPV reported from GenBank,the primers are designed. By PCR,the PCR products are obtained which have fluorescence 1abe- ling,then genechip cross,the genechip detection method is established, and its sensitivity and repeatability detected. [Result] The detection method of CSFV,PRRSV,PCV-2 and PPV by Genechip technique is es- tablished,and the result of detecting the samples from clinical shows that there are 25 CSFV,45 PRRSV, 70 PCV-2,0 PPV,and 18 mixed infections. [Conclusion] The detection method of CSFV,PRRSV,PCV-2 and PPV by Genechip technique is established. The detection method has better specificity, sensitivity andrepeatability.

关 键 词：猪病 基因芯片 猪瘟病毒 高致病性猪蓝耳病病毒 猪圆环病毒2型 猪细小病毒 

分 类 号：S851.347.101[农业科学—预防兽医学]