应用Nested-PCR技术检测人工感染绵羊蓝舌病病毒  

Application of Nested - PCR Technique for the Detection of BTV Antigen in Serum Samples From Experimentally Infected Sheep

作　　者：马洪超[1] 孙淑芳[1] 陶茂晖 尹燕博[1] 郑文伯 蒋正军[1] 蔡丽娟[1] 

机构地区：[1]农业部动物检疫所,青岛266032 [2]河北农业大学 [3]河北畜牧兽医总站

出　　处：《中国动物检疫》2000年第12期20-22,共3页China Animal Health Inspection

摘　　要：实验采用６只绵羊分成３组，一组作为对照接种ＢＨＫ２１细胞上清液，另二组分别接种ＢＴＶ１０和中国分离株Ｚ１株，攻毒定期采血提取抗原并提取病毒ＲＮＡ，同时进行琼脂扩散试验、普通ＰＣＲ、Ｎｅｓｔｅｄ－ＰＣＲ试验。结果在整个攻毒期间，琼脂扩散方法检测不到抗原，普通 ＰＣＲ方法只能在攻毒后 １２ ｄ～ １５ ｄ测得抗原，而 Ｎｅｓｔｅｄ－ＰＣＲ在攻毒后 ６ ｄ即可测得病毒 ＲＮＡ，并且可一直持续到 ３０ ｄ。试验证明， Ｎｅｓｔｅｄ－ ＰＣＲ方法是高度敏感的检测抗原的方法。在临床上能够较早地检出抗原，这对于及时采取有效措施，控制疾病流行具有积极的意义。sheep were divided into 3 groups. The first group was inoculated with BTV10, the second group with Z1(BTV16) a Chinese local strain, the third group with BHK21 cell tissue culture supernatant as control. Blood samples were collected regularly after inoculation. BTV antigen BTV RMA were extracted from blood. AGID can not defect BTV antigen during the entire experiment. PCR can only detect antigen at 15dpi. Nested - PCR can detect BTV RNA from 6 to 30dpi.The experiment proved that Nested-PCRis a highly sensitive technique for detecting BTV antigen. It can detect BTV antigen earlier on clinical samples. It's important for early detection and prevention of BTV disease transmission.

关 键 词：Nested-PCR技术 检测 人工感染 绵羊 蓝舌病病毒 琼脂扩散试验 

分 类 号：S858.26[农业科学—临床兽医学] S852.659.4[农业科学—兽医学]