GB/T 44613-2024 牛羊副流感病毒3型诊断技术

ICS

11.220

CCS

B41

中华人民共和国国家标准

GB/T44613—2024

牛羊副流感病毒3型诊断技术

Diagnostictechniquesforbovineandcaprineparainfluenzavirustype3

2024-09-29发布2025-04-01实施

国家市场监督管理总局

发布

国家标准化管理委员会

GB/T44613—2024

目次

前言

Ⅲ

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引言

Ⅳ

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范围

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2

规范性引用文件

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3

术语和定义

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4

缩略语

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5

临床诊断

2

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样品采集、处理与保存

2

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7

病毒分离鉴定

4

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RT﹘PCR试验

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Real﹘timeRT﹘PCR试验

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10

cELISA试验

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病毒中和试验

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综合判定

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附录A（规范性）试剂配制

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附录B（规范性）病毒核酸检测用引物和荧光探针序列

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附录C（规范性）RT﹘PCR和Real﹘timeRT﹘PCR用阳性对照品制备、引物及序列

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附录D（规范性）RT﹘PCR和Real﹘timeRT﹘PCR反应体系及反应程序

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附录E（资料性）RT﹘PCR和Real﹘timeRT﹘PCR试验结果图

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附录F（资料性）病毒标准阳性血清、标准阴性血清和抗原包被酶标板的制备

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附录G（资料性）酶标单克隆抗体的制备

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Ⅰ

GB/T44613—2024

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。

请注意本文件的某些内容可能涉及专利。本文件的发布机构不承担识别专利的责任。

本文件由中华人民共和国农业农村部提出。

本文件由全国动物卫生标准化技术委员会（SAC/TC181）归口。

本文件起草单位：吉林农业科技学院、江苏省农业科学院、吉林省动物疫病预防控制中心、吉林大

学、中国动物卫生与流行病学中心。

本文件主要起草人：郭利、孙亮、李文良、张金玉、宋建德、刘茂军、杨蕾蕾、毛立、李基棕、

程悦宁、郝飞、孙敏、王楠、王振军、孙娜、冯晓宇。

Ⅲ

GB/T44613—2024

引言

牛副流感病毒3型（Bovineparainfluenzavirustype3，BPIV3）和山羊副流感病毒3型（Caprine

parainfluenzavirustype3，CPIV3）同属于副黏病毒科（Paramyxoviridae）副黏病毒亚科（Paramyx﹘

ovirinae）呼吸道病毒属（Respirovirus），两者核苷酸同源性接近81%，血清学有交叉反应。2022

年，国际病毒分类委员会（InternationalCommitteeonTaxonomyofViruses，ICTV）将BPIV3和

CPIV3分别重新命名为牛呼吸道病毒3型（BovineRespirovirus3）和山羊呼吸道病毒3型（Caprine

Respirovirus3），考虑到我国相关标准的延续性，为便于使用者查询和应用，本文件仍采用原病毒命名

名称。

BPIV3主要感染牛，也可能感染鹿、猪、羊等；CPIV3主要感染山羊和绵羊。易感动物感染病毒后

均以发热、咳嗽、流鼻液为主要临床症状，确诊需进行实验室诊断。本文件给出了BPIV3和CPIV3感

染的临床诊断、病原学检测（病毒分离鉴定、RT﹘PCR、Real﹘timeRT﹘PCR）、血清学检测（cEL﹘

ISA和病毒中和试验）的操作规范。在对牛源样品进行病原学检测时，推荐采用RT﹘PCR或Real﹘time

RT﹘PCR方法检测BPIV3；对羊源样品进行病原学检测时，推荐采用RT﹘PCR或Real﹘timeRT﹘PCR方

法同时检测CPIV3和BPIV3。两种病毒在血清学有交叉反应，因此，本文件中的血清学方法能同时检

测CPIV3和BPIV3感染的抗体。

Ⅳ

GB/T44613—2024

牛羊副流感病毒3型诊断技术

1范围

本文件描述了牛副流感病毒3型感染和山羊副流感病毒3型感染的临床诊断、病毒分离鉴定、

RT﹘PCR检测方法、实时荧光RT﹘PCR检测方法、竞争酶联免疫吸附试验和病毒中和试验的诊断

方法。

本文件适用于牛副流感病毒3型感染和山羊副流感病毒3型感染的诊断与检测。

2规范性引用文件

下列文件中的内容通过文中的规范性引用而构成本文件必不可少的条款。其中，注日期的引用文

件，仅该日期对应的版本适用于本文件；不注日期的引用文件，其最新版本（包括所有的修改单）适用

于本文件。

NY/T541兽医诊断样品采集、保存与运输技术规范

3术语和定义

本文件没有需要界定的术语和定义。

4缩略语

下列缩略语适用于本文件。

BPIV3：牛副流感病毒3型（Bovineparainfluenzavirustype3）

cELISA：竞争酶联免疫吸附试验（Competitiveenzyme﹘linkedimmunosorbentassay）

CPE：细胞病变效应（Cytopathiceffect）

CPIV3：山羊副流感病毒3型（Caprineparainfluenzavirustype3）

Ct值：循环阈值（Cyclethreshold）

DEPC：焦碳酸二乙酯（Diethyloxydiformate）

ELISA：酶联免疫吸附试验（Enzyme﹘linkedimmunosorbentassay）

FBS：胎牛血清（Fetalbovineserum）

FITC：异硫氰酸荧光素（Fluoresceinisothiocyanat）

HAT：选择培养基（Hypoxanthine次黄嘌呤，Aminopterin氨基蝶呤，Thymidine胸腺嘧啶核苷）

HRP：辣根过氧化物酶（Horseradishperoxidase）

MDBK细胞：牛肾细胞（Madindarbybovinekidneycell）

MEM：基础培养基（Minimumessentialmedium）

MOI：感染复数（Multiplicityofinfection）

OD值：光密度（Opticaldensity）

PBS：磷酸盐缓冲液（Phosphatebufferedsaline）

PBST：洗涤缓冲液（Phosphatebufferedsalinewithtween20）

PIV3：副流感病毒3型（Parainfluenzavirustype3）

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