GB/T 45105-2024綿羊痘和山羊痘診斷技術(shù)

ICS11.220

CCSB41

中華人民共和國(guó)國(guó)家標(biāo)準(zhǔn)

GB/T45105—2024

綿羊痘和山羊痘診斷技術(shù)

Diagnostictechniquesforsheeppoxandgoatpox

2024?12?31發(fā)布2025?07?01實(shí)施

國(guó)家市場(chǎng)監(jiān)督管理總局

國(guó)家標(biāo)準(zhǔn)化管理委員會(huì)發(fā)布

GB/T45105—2024

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、保存與運(yùn)輸································································································2

7病毒分離與鑒定········································································································3

8組織學(xué)檢查··············································································································4

9PCR檢測(cè)················································································································4

10實(shí)時(shí)熒光PCR檢測(cè)··································································································6

11LAMP檢測(cè)············································································································7

12病毒中和試驗(yàn)·········································································································8

13ELISA··················································································································9

14綜合判定··············································································································10

附錄A（規(guī)范性）常用試劑溶液配制················································································11

附錄B（規(guī)范性）PCR溶液的配制··················································································12

附錄C（資料性）CaPVPCR檢測(cè)結(jié)果對(duì)照·······································································15

附錄D（資料性）CaPV實(shí)時(shí)熒光PCR檢測(cè)結(jié)果對(duì)照··························································16

附錄E（資料性）CaPVLAMP檢測(cè)結(jié)果參照?qǐng)D·································································17

附錄F（資料性）Reed?Muench法計(jì)算中和抗體效價(jià)···························································18

附錄G（資料性）CaPV重組P32蛋白抗原的制備······························································19

附錄H（資料性）陰性對(duì)照和陽性對(duì)照的制備····································································20

附錄I（規(guī)范性）酶聯(lián)免疫吸附試驗(yàn)試劑的配制··································································21

參考文獻(xiàn)····················································································································22

Ⅰ

GB/T45105—2024

前言

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

定起草。

請(qǐng)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(jī)構(gòu)不承擔(dān)識(shí)別專利的責(zé)任。

本文件由中華人民共和國(guó)農(nóng)業(yè)農(nóng)村部提出。

本文件由全國(guó)動(dòng)物衛(wèi)生標(biāo)準(zhǔn)化技術(shù)委員會(huì)（SAC/TC181）歸口。

本文件起草單位：中國(guó)動(dòng)物疫病預(yù)防控制中心、青海省動(dòng)物疫病預(yù)防控制中心、大連民族大學(xué)、青

海省牦牛繁育推廣服務(wù)中心、中國(guó)檢驗(yàn)檢疫科學(xué)研究院、河北農(nóng)業(yè)大學(xué)、新疆維吾爾自治區(qū)動(dòng)物疾病預(yù)

防控制中心、新疆生產(chǎn)建設(shè)兵團(tuán)動(dòng)物疫病預(yù)防控制中心、西藏自治區(qū)動(dòng)物疫病預(yù)防控制中心、四川省動(dòng)

物疫病預(yù)防控制中心、黑龍江省動(dòng)物疫病預(yù)防與控制中心、江西省動(dòng)物疫病預(yù)防控制中心、海南省動(dòng)物

疫病預(yù)防控制中心、北京億森寶生物科技有限公司、山東省濱州畜牧獸醫(yī)研究院、西寧市動(dòng)物疫病預(yù)防

控制中心、重慶市動(dòng)物疫病預(yù)防控制中心、山東省動(dòng)物疫病預(yù)防與控制中心、淄博市博山區(qū)畜牧漁業(yè)服

務(wù)中心、北京市動(dòng)物疫病預(yù)防控制中心、陜西省動(dòng)物疫病預(yù)防控制中心、哈密市動(dòng)物疫病預(yù)防控制中

心、烏魯木齊市動(dòng)物疾病控制與診斷中心、伊犁州動(dòng)物疾病控制與診斷中心、廣西壯族自治區(qū)動(dòng)物疫病

預(yù)防控制中心、成都海關(guān)技術(shù)中心、山西省動(dòng)物疫病預(yù)防控制中心、河北省動(dòng)物疫病預(yù)防控制中心。

本文件主要起草人：孫雨、宋曉暉、曹際娟、李琦、翟新驗(yàn)、肖穎、顧小雪、王傳彬、楊林、蔡金山、

景建武、闞威、袁萬哲、陽愛國(guó)、李盛瓊、高露、蘇貴成、胡廣衛(wèi)、鄭思思、孟茹、王淑琴、甘平、馮春燕、

趙儉波、鐘映梅、張旭、楊濤、德吉玉珍、格桑央宗、種新婷、居馬別克·夏拉巴依、馬曉艷、楊夷平、

李慶霞、遲慶安、陳榮貴、王濤、孫航、劉穎昳、徐琦、畢一鳴、王新杰、孫曉明、高姍姍、白雪冬、徐鵬、

王金良、沈志強(qiáng)、曾政、駱璐、蘭鄒然、張?jiān)?、趙浩軍、李棟梁、趙光明、施開創(chuàng)、莫?jiǎng)偬m、林華、雷宇平、

許玉靜。

Ⅲ

GB/T45105—2024

引言

綿羊痘和山羊痘（以下簡(jiǎn)稱為羊痘）分別由綿羊痘病毒（Sheeppoxvirus，SPV）和山羊痘病毒

（Goatpoxvirus，GPV）引起。綿羊痘病毒和山羊痘病毒均屬于痘病毒科（Poxviridae）、脊椎動(dòng)物痘病毒

亞科（Chordopoxvirdae）、山羊痘病毒屬病毒（Capripoxvirus，CaPV）。我國(guó)將其列為二類動(dòng)物疫病，同

時(shí)也為世界動(dòng)物衛(wèi)生組織（WOAH）名錄疫病。感染動(dòng)物表現(xiàn)為發(fā)熱，皮膚、黏膜、器官表面廣泛性丘

疹或結(jié)節(jié)，皮膚水腫，淋巴結(jié)腫大，消瘦，產(chǎn)乳量大幅度降低，嚴(yán)重時(shí)導(dǎo)致死亡。山羊痘最早于公元前

200年在歐洲發(fā)現(xiàn)，綿羊痘最早于13世紀(jì)在英國(guó)發(fā)現(xiàn)，發(fā)病率50%~80%，病死率20%~80%，其中羔

羊致死率可達(dá)100%。世界各國(guó)均有羊痘發(fā)病與流行的相關(guān)報(bào)道，目前非洲北部、中東和亞洲的部分

國(guó)家流行較為嚴(yán)重。羊痘是阻礙引進(jìn)綿羊和山羊珍貴品種以及養(yǎng)羊業(yè)規(guī)?；a(chǎn)的主要障礙，嚴(yán)重危

害養(yǎng)羊業(yè)生產(chǎn)。羊痘的臨床癥狀易與羊偽結(jié)核和羊傳染性膿皰混淆，對(duì)其進(jìn)行實(shí)驗(yàn)室鑒別診斷。

本文件參考了《陸生動(dòng)物診斷試驗(yàn)與疫苗手冊(cè)》（WOAH）的有關(guān)內(nèi)容，技術(shù)方法與WOAH推薦的

標(biāo)準(zhǔn)方法一致。

Ⅳ

GB/T45105—2024

綿羊痘和山羊痘診斷技術(shù)

1范圍

本文件描述了綿羊痘和山羊痘的臨床診斷、病毒分離與鑒定、PCR檢測(cè)、實(shí)時(shí)熒光PCR檢測(cè)、

LAMP檢測(cè)、病毒中和試驗(yàn)、ELISA等診斷方法。

本文件適用于羊痘的臨床診斷、實(shí)驗(yàn)室檢測(cè)、流行病學(xué)調(diào)查以及檢驗(yàn)檢疫。

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)定性引用而構(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(duì)應(yīng)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。

GB19489實(shí)驗(yàn)室生物安全通用要求

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。

CaPV：山羊痘病毒屬病毒（capripoxvirus）

CPE：細(xì)胞病變效應(yīng)（cytopathiceffect）

Ct值：循環(huán)閾值（cyclethreshold）