ICS11.220

CCSB41

中華人民共和國國家標準

GB/T27530—2025

代替GB/T27530—2011

牛出血性敗血癥診斷技術(shù)

Diagnostictechniquesforbovinehaemorrhagicsepticaemia

2025?01?24發(fā)布2025?08?01實施

國家市場監(jiān)督管理總局

國家標準化管理委員會發(fā)布

GB/T27530—2025

目次

前言··························································································································Ⅲ

引言··························································································································Ⅳ

1范圍·······················································································································1

2規(guī)范性引用文件········································································································1

3術(shù)語和定義··············································································································1

4縮略語····················································································································1

5臨床診斷·················································································································1

6樣品采集、運輸與保存································································································2

7細菌分離與鏡檢········································································································3

8細菌鑒定·················································································································4

9多重聚合酶鏈式反應··································································································8

10綜合判定··············································································································10

附錄A（規(guī)范性）培養(yǎng)基的配制······················································································11

附錄B（規(guī)范性）試劑溶液及電泳介質(zhì)的配制····································································13

附錄C（規(guī)范性）多殺性巴氏桿菌分型抗原、抗血清及綿羊紅細胞的制備方法···························16

附錄D（資料性）瓊脂凝膠免疫擴散試驗打孔及結(jié)果判定示意圖············································17

附錄E（資料性）PCR陽性對照的制備············································································18

附錄F（資料性）多殺性巴氏桿菌檢測引物及擴增序列信息··················································19

附錄G（資料性）多殺性巴氏桿菌多重PCR陽性擴增參照圖················································21

參考文獻····················································································································22

Ⅰ

GB/T27530—2025

前言

本文件按照GB/T1.1—2020《標準化工作導則第1部分：標準化文件的結(jié)構(gòu)和起草規(guī)則》的規(guī)

定起草。

本文件代替GB/T27530—2011《牛出血性敗血病診斷技術(shù)》，與GB/T27530—2011相比，除結(jié)構(gòu)

調(diào)整和編輯性改動外，主要技術(shù)變化如下：

——更改了適用范圍（見第1章，2011年版的第1章）；

——增加了縮略語（見第4章）；

——更改了臨床診斷特點，并增加了臨床診斷的結(jié)果判定（見第5章，2011年版的第4章）；

——更改了細菌分離、鏡檢、鑒定的操作步驟（見第7章、第8章，2011年版的第5章）；

——增加了細菌的分離培養(yǎng)所用培養(yǎng)基（見7.1、附錄A）；

——更改了細菌的涂片染色鏡檢（見7.2、附錄B，2011年版的5.2、附錄A）；

——刪除了MR、VP試驗，β?半乳糖苷酶試驗，檸檬酸鹽利用試驗（見2011年版的5.3.2.3.2、

5.3.2.3.4、5.3.2.3.5）；

——更改了細菌的生化特性鑒定方法，并規(guī)定了不同生化試驗的培養(yǎng)基制備（見8.1、附錄A、附錄

B，2011年版的5.3.2.3）；

——更改了間接血凝試驗的莢膜抗原與抗血清制備方法（見8.2，2011年版的5.3.2.4.1）；

——增加了莢膜型鑒定的判定標準和多殺性巴氏桿菌的多重PCR鑒定技術(shù)（見第9章）；

——更改了綜合判定標準（見第10章，2011年版的第6章）。

請注意本文件的某些內(nèi)容可能涉及專利。本文件的發(fā)布機構(gòu)不承擔識別專利的責任。

本文件由中華人民共和國農(nóng)業(yè)農(nóng)村部提出。

本文件由全國動物衛(wèi)生標準化技術(shù)委員會（SAC/TC181）歸口。

本文件起草單位：北京市農(nóng)林科學院、中國動物衛(wèi)生與流行病學中心、甘肅農(nóng)業(yè)大學、福建省農(nóng)業(yè)

科學院畜牧獸醫(yī)研究所。

本文件主要起草人：李永清、宋翠平、劉文曉、胡永浩、秦立得、趙思俊、王玉東、傅光華、黃瑜、

曹旭敏、李木子、程龍飛、隋金鈺、江波、程晶、徐福洲、周林宜、王曉茵、孫曉亮、王淑婷、劉坤、張柏銘。

本文件及其所代替文件的歷次版本發(fā)布情況為：

——2011年首次發(fā)布為GB/T27530—2011；

——本次為第一次修訂。

Ⅲ

GB/T27530—2025

引言

牛出血性敗血癥（Bovinehaemorrhagicsepticaemia）是黃牛、水牛、牦牛、奶牛和野牛的一種主要疫

病，由多殺性巴氏桿菌（Pasteurellamultocida，Pm）某些血清型引起，典型癥狀為高致死性急性敗血癥

和纖維素性化膿性肺炎等。本病以散發(fā)為主，偶爾呈小區(qū)域地方流行，危害較大。世界動物衛(wèi)生組織

（WOAH）將牛出血性敗血癥列為法定名錄疫病，我國將其列為三類動物疫病。

多殺性巴氏桿菌分為5個莢膜血清型（A、B、D、E、F），各莢膜血清型之間的交叉免疫保護作用弱，

制訂針對性防控措施時須首先進行血清型鑒定。

本文件的修訂參考了WOAH《陸生動物診斷試驗和疫苗手冊》（2022），并結(jié)合了我國相關(guān)技術(shù)研

究新成果。

Ⅳ

GB/T27530—2025

牛出血性敗血癥診斷技術(shù)

1范圍

本文件描述了牛出血性敗血癥的臨床診斷、樣品采集、運輸與保存，以及多殺性巴氏桿菌的分離培

養(yǎng)、染色鏡檢、生化試驗、聚合酶鏈式反應等實驗室診斷方法。

本文件適用于牛出血性敗血癥的診斷。

2規(guī)范性引用文件

下列文件中的內(nèi)容通過文中的規(guī)范性引用而構(gòu)成本文件必不可少的條款。其中，注日期的引用文

件，僅該日期對應的版本適用于本文件；不注日期的引用文件，其最新版本（包括所有的修改單）適用于

本文件。

NY/T541獸醫(yī)診斷樣本采集、保存與運輸技術(shù)規(guī)范

3術(shù)語和定義

本文件沒有需要界定的術(shù)語和定義。

4縮略語

下列縮略語適用于本文件：

AGID：瓊脂凝膠免疫擴散試驗（agargelimmunediffusiontest）

CIEP：對流免疫電泳試驗（counterimmunoelec