T/CVDA 1-2022 动物西尼罗病毒中和抗体检测技术

ICS11.220

B41

T/CVDA

团体标准

T/CVDA1—2022

动物西尼罗病毒中和抗体检测技术

DetectingtechniqueofneutralizationantibodyagainstWestNilevirusinanimals

2022-07-19发布2022-07-25实施

中国兽药协会发布

T/CVDA1—2022

目次

前言................................................................................II

1范围...............................................................................1

2规范性引用文件.....................................................................1

3缩略语.............................................................................1

4试剂和材料.........................................................................1

5器材和设备.........................................................................2

6动物血清的采集.....................................................................2

7PRNT实验操作.......................................................................2

7.1安全防护.......................................................................2

7.2材料准备.......................................................................2

7.2.1细胞......................................................................2

7.2.2待测血清..................................................................2

7.2.3检测用细胞板..............................................................2

7.3检测操作.......................................................................3

7.3.1血清的稀释................................................................3

7.3.2病毒与血清的中和..........................................................3

7.3.3接种细胞..................................................................3

7.3.4蚀斑计数..................................................................3

8结果观察与判定.....................................................................4

8.1观察顺序.......................................................................4

8.2蚀斑观察结果判定...............................................................4

8.2.1试验成立条件...............................................................4

8.2.2结果判定与效价计算........................................................4

A.1材料和试剂.......................................................................1

A.2方法.............................................................................1

A.2.1表达质粒构建.................................................................1

A.2.2细胞转染与克隆筛选...........................................................1

A.2.3克隆细胞株的鉴定.............................................................1

B.1材料和试剂.......................................................................1

B.2方法.............................................................................1

B.2.1WNV复制子质粒构建............................................................1

B.2.2△WNV病毒的初次包装..........................................................1