2021年实训十三凝集试验.pdf

1实训十三凝集试验目的要求掌握平板凝集试验和试管凝集试验的操作及结果判定。明确凝集试验在生产中的应用。仪器及材料 试管架、凝集管、吸管(5ml、1ml、0.5ml)、生理盐水、玻板、25卩l 移液器(配带滴头)、被检血清、温箱、鸡白痢全血平板凝集抗原、鸡白痢阳性血清、鸡白痢阴性血清、布氏杆菌平板凝集抗原和试管凝集抗原，布氏杆菌标准阳性血清和阴性血清，羊布氏杆菌病待检血清。方法与步骤(一)鸡白痢快速全血平板凝集试验(实验视频十 )1.取一块洁净的玻板或白瓷板，划出3X 3cm的方块。2?在每个方块的中心滴一滴(约0.02ml)鸡白痢全血平板凝集抗原。3.用采血针刺破鸡冠或翅静脉采集新鲜血液。4.在抗原滴旁边滴加等量的一滴新鲜血液。5.用一洁净的细玻璃棒或牙签将抗原与血液混匀，不同检样不能用同一牙签混匀。6.轻轻摇动，使混合物反应2分钟。结果 室温下，在2分钟内出现凝集现象者为阳性；2分钟内未出现凝集现象者为阴性。注意：每次试验都要设阳性和阴性对照，做法同全血平板凝集试验。对可疑的鸡应根据鸡群以往感染情况做出解释，如鸡群以前为阳性群，则可疑反应应判为阳性。新近感染的鸡，还需在3?4周后作重新试验。(二)布氏杆菌病的凝集试验1.布氏杆菌病的平板凝集试验(1)取一长方形洁净玻璃板，用玻璃铅笔划成若干方格，每格约4cm2,编号。(2)分别吸取25卩l被检血清、生理盐水、标准阳性血清、标准阴性血清加于不同编号的方格内，每吸一种成分需换一个滴头。然后吸取25卩l抗原加到玻璃板的每一个格内，用牙签将抗原与血清充分混匀，每混匀一格需更换一根牙签。于室温(15 C以上)静置4min观察结果，如室温过低，可适当加温。(3)在对照标准阳性血清(+)、标准阴性血清(-)、生理盐水(-)反应正常的前提下，被检血清出现大的凝集片或小的颗粒状物，液体透明判为阳性(+)，液体均匀浑浊，无任何凝集物判为阴性(-)。本法操作简便，容易掌握和判断，适用于大群家畜的检疫。筛选出的阳性反应血清，再做试管凝集试验，以试管凝集的结果为被检血清的最终结果。2.布氏杆菌病的试管凝集试验(1)每份血清用5支试管，另取对照管3支，置于试管架上。如待检血清多时，对照只需做一份。(2)按实表7取0.5%石炭酸生理盐水(检绵羊和山羊血清，则用10盐水稀释血清和抗原)第1孔加入2.3ml，2?5孔加入0.5ml，然后另取吸管吸取被检血清0.2ml，加入第1管中，并反复吸吹3次将血清与生理盐水充分混合均匀，吸出1.5ml弃之，再吸出0.5ml 加入第2管，混精品w o r d 学习资料 可编辑资料-精心整理-欢迎下载-第 1 页，共 3 页2匀后吸出0.5ml 加入第3管，依此类推至第5管，混匀后吸出0.5ml 弃去。精品w o r d 学习资料 可编辑资料-精心整理-欢迎下载-第 2 页，共 3 页文档编码:CW4F4S8C2I9 HO5D6M4N2M7 ZQ7U1L1G5K5文档编码:CW4F4S8C2I9 HO5D6M4N2M7 ZQ7U1L1G5K5文档编码:CW4F4S8C2I9 HO5D6M4N2M7 ZQ7U1L1G5K5文档编码:CW4F4S8C2I9 HO5D6M4N2M7 ZQ7U1L1G5K5文档编码:CW4F4S8C2I9 HO5D6M4N2M7 ZQ7U1L1G5K5文档编码:CW4F4S8C2I9 HO5D6M4N2M7 ZQ7U1L1G5K5文档编码:CW4F4S8C2I9 HO5D6M4N2M7 ZQ7U1L1G5K5文档编码:CW4F4S8C2I9 HO5D6M4N2M7 ZQ7U1L1G5K5文档编码:CW4F4S8C2I9 HO5D6M4N2M7 ZQ7U1L1G5K5文档编码:CW4F4S8C2I9 HO5D6M4N2M7 ZQ7U1L1G5K5文档编码:CW4F4S8C2I9 HO5D6M4N2M7 ZQ7U1L1G5K5文档编码:CW4F4S8C2I9 HO5D6M4N2M7 ZQ7U1L1G5K5文档编码:CW4F4S8C2I9 HO5D6M4N2M7 ZQ7U1L1G5K5文档编码:CW4F4S8C2I9 HO5D6M4N2M7 ZQ7U1L1G5K5文档编码:CW4F4S8C2I9 HO5D6M4N2M7 ZQ7U1L1G5K5文档编码:CW4F4S8C2I9 HO5D6M4N2M7 ZQ7U1L1G5K5文档编码:CW4F4S8C2I9 HO5D6M4N2M7 ZQ7U1L1G5K5文档编码:CW4F4S8C2I9 HO5D6M4N2M7 ZQ7U1L1G5K5文档编码:CW4F4S8C2I9 HO5D6M4N2M7 ZQ7U1L1G5K5文档编码:CW4F4S8C2I9 HO5D6M4N2M7 ZQ7U1L1G5K5文档编码:CW4F4S8C2I9 HO5D6M4N2M7 ZQ7U1L1G5K5文档编码:CW4F4S8C2I9 HO5D6M4N2M7 ZQ7U1L1G5K5文档编码:CW4F4S8C2I9 HO5D6M4N2M7 ZQ7U1L1G5K5文档编码:CW4F4S8C2I9 HO5D6M4N2M7 ZQ7U1L1G5K5文档编码:CW4F4S8C2I9 HO5D6M4N2M7 ZQ7U1L1G5K5文档编码:CW4F4S8C2I9 HO5D6M4N2M7 ZQ7U1L1G5K5文档编码:CW4F4S8C2I9 HO5D6M4N2M7 ZQ7U1L1G5K5文档编码:CW4F4S8C2I9 HO5D6M4N2M7 ZQ7U1L1G5K5文档编码:CW4F4S8C2I9 HO5D6M4N2M7 ZQ7U1L1G5K5文档编码:CW4F4S8C2I9 HO5D6M4N2M7 ZQ7U1L1G5K5文档编码:CW4F4S8C2I9 HO5D6M4N2M7 ZQ7U1L1G5K5文档编码:CW4F4S8C2I9 HO5D6M4N2M7 ZQ7U1L1G5K5文档编码:CW4F4S8C2I9 HO5D6M4N2M7 ZQ7U1L1G5K5文档编码:CW4F4S8C2I9 HO5D6M4N2M7 ZQ7U1L1G5K5文档编码:CW4F4S8C2I9 HO5D6M4N2M7 ZQ7U1L1G5K5文档编码:CW4F4S8C2I9 HO5D6M4N2M7 ZQ7U1L1G5K5文档编码:CW4F4S8C2I9 HO5D6M4N2M7 ZQ7U1L1G5K5文档编码:CW4F4S8C2I9 HO5D6M4N2M7 ZQ7U1L1G5K5文档编码:CW4F4S8C2I9 HO5D6M4N2M7 ZQ7U1L1G5K5文档编码:CW4F4S8C2I9 HO5D6M4N2M7 ZQ7U1L1G5K5文档编码:CW4F4S8C2I9 HO5D6M4N2M7 ZQ7U1L1G5K5文档编码:CW4F4S8C2I9 HO5D6M4N2M7 ZQ7U1L1G5K5文档编码:CW4F4S8C2I9 HO5D6M4N2M7 ZQ7U1L1G5K5文档编码:CW4F4S8C2I9 HO5D6M4N2M7 ZQ7U1L1G5K5文档编码:CW4F4S8C2I9 HO5D6M4N2M7 ZQ7U1L1G5K5文档编码:CW4F4S8C2I9 HO5D6M4N2M7 ZQ7U1L1G5K5文档编码:CW4F4S8C2I9 HO5D6M4N2M7 ZQ7U1L1G5K5文档编码:CW4F4S8C2I9 HO5D6M4N2M7 ZQ7U1L1G5K53（3）将布氏杆菌试管凝集抗原用0.5%石炭酸生理盐水稀释20倍，在各管中分别加入的0.5ml，加完后，充分振荡，放入37C温箱中24h，取出观察并记录结果。（4）结果判定判定结果时用“+”表示反应的强度。以出现“+”以上凝集现象的最高稀释倍数，作为该血清的凝集价（滴度）。“+”：液体完全透明，菌体完全被凝集呈伞状沉于管底，振荡时，沉淀物呈片状、块状或颗粒状，表示菌体100%凝集。“+”：液体稍有浑浊，菌体大部分被凝集沉于管底，振荡时情况同上，表示菌体75%凝集。“+”：液体呈淡乳白色浑浊，管底有明显的凝集沉淀，振荡时有块状或小片絮状物，表示菌体50%凝集。“+”：液体浑浊，不透明，管底有少许凝集沉淀，表示菌体25%凝集。“-”：液体完全浑浊，不透明，有时管底中央有一部分圆点状沉淀物，振荡时均匀浑浊，表示菌体完全不凝集。确定血清凝集价（滴度）时，应以出现+以上凝集现象的最高稀释度为准。（5）判定标准 牛、马和骆驼凝集价为1:100，猪、羊和狗的凝集价为1:50判为阳 性。牛、马和骆驼凝集价为1:50，猪、羊和狗凝集价为1:25判为可疑。可疑反应的家畜，经3?4周后再采血重新检查，牛和羊仍为可疑判为阳性。猪和马 仍为可疑，而畜群中又没有病例和大批阳性病畜，则判为阴性。六、注意事项1?被检血清必须新鲜，无溶血现象和腐败气味。2?每次需做阳性血清、阴性血清和抗原三种对照试管号12345678血清稀释倍数1:12.51:251:501:1001:200对照抗原对照阳性血清1:25阴性血清1:252.30.50.5 0.2?0.5-J 0.50.50.50.5弃去 1.5第6管中不加血清，第7管中加1:25稀释的布氏杆菌阳性血清0.5ml，第8管中加1:25稀释 的布氏杆菌阴性血清0.5ml（取阳性血清、阴性血清应另换吸管）。实表 7 布氏杆菌试管凝集试验术式表0.5%石炭酸生理盐水（ml）被检血清（ml）1:20 抗原（ml）精品w o r d 学习资料 可编辑资料-精心整理-欢迎下载-第 3 页，共 3 页文档编码:CW4F4S8C2I9 HO5D6M4N2M7 ZQ7U1L1G5K5文档编码:CW4F4S8C2I9 HO5D6M4N2M7 ZQ7U1L1G5K5文档编码:CW4F4S8C2I9 HO5D6M4N2M7 ZQ7U1L1G5K5文档编码:CW4F4S8C2I9 HO5D6M4N2M7 ZQ7U1L1G5K5文档编码:CW4F4S8C2I9 HO5D6M4N2M7 ZQ7U1L1G5K5文档编码:CW4F4S8C2I9 HO5D6M4N2M7 ZQ7U1L1G5K5文档编码:CW4F4S8C2I9 HO5D6M4N2M7 ZQ7U1L1G5K5文档编码:CW4F4S8C2I9 HO5D6M4N2M7 ZQ7U1L1G5K5文档编码:CW4F4S8C2I9 HO5D6M4N2M7 ZQ7U1L1G5K5文档编码:CW4F4S8C2I9 HO5D6M4N2M7 ZQ7U1L1G5K5文档编码:CW4F4S8C2I9 HO5D6M4N2M7 ZQ7U1L1G5K5文档编码:CW4F4S8C2I9 HO5D6M4N2M7 ZQ7U1L1G5K5文档编码:CW4F4S8C2I9 HO5D6M4N2M7 ZQ7U1L1G5K5文档编码:CW4F4S8C2I9 HO5D6M4N2M7 ZQ7U1L1G5K5文档编码:CW4F4S8C2I9 HO5D6M4N2M7 ZQ7U1L1G5K5文档编码:CW4F4S8C2I9 HO5D6M4N2M7 ZQ7U1L1G5K5文档编码:CW4F4S8C2I9 HO5D6M4N2M7 ZQ7U1L1G5K5文档编码:CW4F4S8C2I9 HO5D6M4N2M7 ZQ7U1L1G5K5文档编码:CW4F4S8C2I9 HO5D6M4N2M7 ZQ7U1L1G5K5文档编码:CW4F4S8C2I9 HO5D6M4N2M7 ZQ7U1L1G5K5文档编码:CW4F4S8C2I9 HO5D6M4N2M7 ZQ7U1L1G5K5文档编码:CW4F4S8C2I9 HO5D6M4N2M7 ZQ7U1L1G5K5文档编码:CW4F4S8C2I9 HO5D6M4N2M7 ZQ7U1L1G5K5文档编码:CW4F4S8C2I9 HO5D6M4N2M7 ZQ7U1L1G5K5文档编码:CW4F4S8C2I9 HO5D6M4N2M7 ZQ7U1L1G5K5文档编码:CW4F4S8C2I9 HO5D6M4N2M7 ZQ7U1L1G5K5文档编码:CW4F4S8C2I9 HO5D6M4N2M7 ZQ7U1L1G5K5文档编码:CW4F4S8C2I9 HO5D6M4N2M7 ZQ7U1L1G5K5文档编码:CW4F4S8C2I9 HO5D6M4N2M7 ZQ7U1L1G5K5文档编码:CW4F4S8C2I9 HO5D6M4N2M7 ZQ7U1L1G5K5文档编码:CW4F4S8C2I9 HO5D6M4N2M7 ZQ7U1L1G5K5文档编码:CW4F4S8C2I9 HO5D6M4N2M7 ZQ7U1L1G5K5文档编码:CW4F4S8C2I9 HO5D6M4N2M7 ZQ7U1L1G5K5文档编码:CW4F4S8C2I9 HO5D6M4N2M7 ZQ7U1L1G5K5文档编码:CW4F4S8C2I9 HO5D6M4N2M7 ZQ7U1L1G5K5文档编码:CW4F4S8C2I9 HO5D6M4N2M7 ZQ7U1L1G5K5文档编码:CW4F4S8C2I9 HO5D6M4N2M7 ZQ7U1L1G5K5文档编码:CW4F4S8C2I9 HO5D6M4N2M7 ZQ7U1L1G5K5文档编码:CW4F4S8C2I9 HO5D6M4N2M7 ZQ7U1L1G5K5文档编码:CW4F4S8C2I9 HO5D6M4N2M7 ZQ7U1L1G5K5文档编码:CW4F4S8C2I9 HO5D6M4N2M7 ZQ7U1L1G5K5文档编码:CW4F4S8C2I9 HO5D6M4N2M7 ZQ7U1L1G5K5文档编码:CW4F4S8C2I9 HO5D6M4N2M7 ZQ7U1L1G5K5文档编码:CW4F4S8C2I9 HO5D6M4N2M7 ZQ7U1L1G5K5文档编码:CW4F4S8C2I9 HO5D6M4N2M7 ZQ7U1L1G5K5文档编码:CW4F4S8C2I9 HO5D6M4N2M7 ZQ7U1L1G5K5文档编码:CW4F4S8C2I9 HO5D6M4N2M7 ZQ7U1L1G5K5文档编码:CW4F4S8C2I9 HO5D6M4N2M7 ZQ7U1L1G5K5