DB53/T 1059.1-2021 滇金丝猴检疫技术 第1部分：产气荚膜梭菌实验室检测技术规范

ICS11.220

CCSB41

53

云南省地方标准

DB53/T1059.1—2021

滇金丝猴检疫技术第1部分：产气荚膜

梭菌实验室检测技术规范

QuarantinetechniqueforRhinopithecusbieti

—Part1:Technicalspecificationsforlaboratoryexaminationof

Clostridiumperfringens

2021-09-30发布2021-10-14实施

云南省市场监督管理局发布

DB53/T1059.1—2021

目  次

前言.....................................................................................................................................................................III

引言.....................................................................................................................................................................Ⅴ

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

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

3术语和定义.....................................................................................................................................................1

4缩略语.............................................................................................................................................................2

5总则.................................................................................................................................................................2

6产气荚膜梭菌分型.........................................................................................................................................3

7仪器设备和试剂.............................................................................................................................................3

7.1采样用品..................................................................................................................................................3

7.2实验材料..................................................................................................................................................3

7.3实验设备..................................................................................................................................................3

8样品.................................................................................................................................................................3

8.1采样要求..................................................................................................................................................4

8.2采样程序..................................................................................................................................................4

8.3样品保存与运输......................................................................................................................................4

9试验步骤.........................................................................................................................................................4

9.1细菌分离培养及培养特性观察..............................................................................................................4

9.2革兰染色和芽孢染色镜检......................................................................................................................4

9.3细菌生化试验..........................................................................................................................................4

9.4小鼠接种试验..........................................................................................................................................4

9.516SrRNA基因与4种毒素基因的PCR扩增及测序分析...............................................................5

10试验数据处理...............................................................................................................................................5

10.1细菌培养结果判定................................................................................................................................5

10.2染色镜检结果判读................................................................................................................................5

10.3生化试验结果判定................................................................................................................................6

10.4小鼠毒性试验结果判定........................................................................................................................6

10.5毒素中和试验结果判定........................................................................................................................6

10.6核酸检测结果判定................................................................................................................................6

附录A（规范性）引物和16SrRNA基因PCR扩增体系.......................................................................7

附录B（规范性）采样记录表.......................................................................................................................8

附录C（规范性）革兰染色和雪-浮染色方法.............................................................................................9

附录D（规范性）TA克隆程序...................................................................................................................10

I

DB53/T1059.1—2021

前  言

本文件按照GB/T1.1-2020《标准化工作导则第1部分：标准化文件的结构和起草规则》的规定起

草。

本文件是DB53/T1059《滇金丝猴检疫技术》的第1部分。DB53/T1059已经发布了以下部分：

—