GB/T 44613-2024牛羊副流感病毒3型診斷技術(shù)

ICS

11.220

CCS

B41

中華人民共和國國家標(biāo)準(zhǔn)

GB/T44613—2024

牛羊副流感病毒3型診斷技術(shù)

Diagnostictechniquesforbovineandcaprineparainfluenzavirustype3

2024-09-29發(fā)布2025-04-01實(shí)施

國家市場監(jiān)督管理總局發(fā)布

國家標(biāo)準(zhǔn)化管理委員會(huì)

GB/T44613—2024

目次

前言

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Ⅲ

引言

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Ⅳ

1

范圍

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規(guī)范性引用文件

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3

術(shù)語和定義

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縮略語

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5

臨床診斷

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6

樣品采集、處理與保存

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7

病毒分離鑒定

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RT?PCR試驗(yàn)

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9

Real?timeRT?PCR試驗(yàn)

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cELISA試驗(yàn)

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病毒中和試驗(yàn)

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綜合判定

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11

附錄A（規(guī)范性）試劑配制

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12

附錄B（規(guī)范性）病毒核酸檢測用引物和熒光探針序列

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13

附錄C（規(guī)范性）RT?PCR和Real?timeRT?PCR用陽性對照品制備、引物及序列

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14

附錄D（規(guī)范性）RT?PCR和Real?timeRT?PCR反應(yīng)體系及反應(yīng)程序

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附錄E（資料性）RT?PCR和Real?timeRT?PCR試驗(yàn)結(jié)果圖

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18

附錄F（資料性）病毒標(biāo)準(zhǔn)陽性血清、標(biāo)準(zhǔn)陰性血清和抗原包被酶標(biāo)板的制備

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附錄G（資料性）酶標(biāo)單克隆抗體的制備

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Ⅰ

GB/T44613—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農(nóng)業(yè)農(nóng)村部提出。

本文件由全國動(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：吉林農(nóng)業(yè)科技學(xué)院、江蘇省農(nóng)業(yè)科學(xué)院、吉林省動(dòng)物疫病預(yù)防控制中心、吉林大

學(xué)、中國動(dòng)物衛(wèi)生與流行病學(xué)中心。

本文件主要起草人：郭利、孫亮、李文良、張金玉、宋建德、劉茂軍、楊蕾蕾、毛立、李基棕、

程悅寧、郝飛、孫敏、王楠、王振軍、孫娜、馮曉宇。

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）同屬于副黏病毒科（Paramyxoviridae）副黏病毒亞科（Paramyx?

ovirinae）呼吸道病毒屬（Respirovirus），兩者核苷酸同源性接近81%，血清學(xué)有交叉反應(yīng)。2022

年，國際病毒分類委員會(huì)（InternationalCommitteeonTaxonomyofViruses，ICTV）將BPIV3和

CPIV3分別重新命名為牛呼吸道病毒3型（BovineRespirovirus3）和山羊呼吸道病毒3型（Caprine

Respirovirus3），考慮到我國相關(guān)標(biāo)準(zhǔn)的延續(xù)性，為便于使用者查詢和應(yīng)用，本文件仍采用原病毒命名

名稱。

BPIV3主要感染牛，也可能感染鹿、豬、羊等；CPIV3主要感染山羊和綿羊。易感動(dòng)物感染病毒后

均以發(fā)熱、咳嗽、流鼻液為主要臨床癥狀，確診需進(jìn)行實(shí)驗(yàn)室診斷。本文件給出了BPIV3和CPIV3感

染的臨床診斷、病原學(xué)檢測（病毒分離鑒定、RT?PCR、Real?timeRT?PCR）、血清學(xué)檢測（cEL?

ISA和病毒中和試驗(yàn)）的操作規(guī)范。在對牛源樣品進(jìn)行病原學(xué)檢測時(shí)，推薦采用RT?PCR或Real?time

RT?PCR方法檢測BPIV3；對羊源樣品進(jìn)行病原學(xué)檢測時(shí)，推薦采用RT?PCR或Real?timeRT?PCR方

法同時(shí)檢測CPIV3和BPIV3。兩種病毒在血清學(xué)有交叉反應(yīng)，因此，本文件中的血清學(xué)方法能同時(shí)檢

測CPIV3和BPIV3感染的抗體。

Ⅳ

GB/T44613—2024

牛羊副流感病毒3型診斷技術(shù)

1范圍

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型感染的臨床診斷、病毒分離鑒定、

RT?PCR檢測方法、實(shí)時(shí)熒光RT?PCR檢測方法、競爭酶聯(lián)免疫吸附試驗(yàn)和病毒中和試驗(yàn)的診斷

方法。

本文件適用于牛副流感病毒3型感染和山羊副流感病毒3型感染的診斷與檢測。

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(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。

NY/T541獸醫(yī)診斷樣品采集、保存與運(yùn)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。

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

cELISA：競爭酶聯(lián)免疫吸附試驗(yàn)（Competitiveenzyme?linkedimmunosorbentassay）

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

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

Ct值：循環(huán)閾值（Cyclethreshold）

DEPC：焦碳酸二乙酯（Diethyloxydiformate）

ELISA：酶聯(lián)免疫吸附試驗(yàn)（Enzyme?linkedimmunosorbentassay）

FBS：胎牛