ICS11.220

CCSB41

中華人民共和國國家標(biāo)準(zhǔn)

GB/T26436—2025

代替GB/T26436—2010

禽白血病診斷技術(shù)

Diagnostictechniquesforavianleukosis

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(biāo)準(zhǔn)化管理委員會發(fā)布

GB/T26436—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實驗室生物安全措施··································································································2

6臨床診斷·················································································································2

6.1流行病學(xué)···········································································································2

6.2臨床癥狀···········································································································2

6.3病理變化···········································································································3

6.4鑒別診斷···········································································································3

6.5臨床判定···········································································································3

7樣品采集與處理········································································································4

7.1樣品采集···········································································································4

7.2樣品處理···········································································································4

8實驗室診斷··············································································································4

8.1病毒分離和鑒定··································································································4

8.2病毒亞群的鑒定··································································································7

8.3Real?timeRT?PCR擴增ALV?J···············································································7

8.4血清學(xué)診斷········································································································7

9綜合判定·················································································································8

9.1疑似·················································································································8

9.2確診·················································································································8

附錄A（規(guī)范性）病毒分離相關(guān)溶液的配制········································································9

A.10.01mol/LPBS（pH7.4）······················································································9

A.250%甘油?PBS保存液（pH7.4）··············································································9

A.3細(xì)胞完全營養(yǎng)液和維持液······················································································9

附錄B（資料性）CEF（C/E品系）的制備··········································································10

附錄C（資料性）抗ALV單因子雞血清的制備··································································11

附錄D（規(guī)范性）p27抗原ELISA檢測相關(guān)溶液的配制·······················································12

D.1包被液············································································································12

D.2洗滌液············································································································12

Ⅰ

GB/T26436—2025

D.3樣品稀釋液······································································································12

D.4底物溶液·········································································································12

D.5終止液············································································································12

附錄E（資料性）ALV亞群鑒定程序···············································································13

E.1克隆載體和宿主菌·····························································································13

E.2病毒模板的制備································································································13

E.3囊膜蛋白env基因的擴增·····················································································13

E.4PCR產(chǎn)物的克隆·······························································································14

E.5質(zhì)粒DNA的提取和鑒定····················································································15

E.6克隆序列的測定································································································15

附錄F（資料性）Real?timeRT?PCR擴增ALV?J································································17

F.1引物和探針序列································································································17

F.2樣品采集和前處理·····························································································17

F.3核酸抽提·········································································································17

F.4擴增試劑準(zhǔn)備···································································································17

F.5加樣檢測·········································································································18

F.6反應(yīng)條件設(shè)置···································································································18

F.7熒光素設(shè)定······································································································18

F.8分析條件設(shè)定及結(jié)果判定····················································································18

附錄G（資料性）ALV?J一步法Real?timeRT?PCR檢測試劑盒的組成及使用···························19

G.1試劑盒組成······································································································19

G.2說明···············································································································19

G.3使用時的注意事項·····························································································19

附錄H（資料性）IFA法抗體檢測用ALV感染細(xì)胞的制備··················································20

參考文獻(xiàn)····················································································································21

Ⅱ

GB/T26436—2025

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。

本文件代替GB/T26436—2010《禽白血病診斷技術(shù)》，與GB/T26436—2010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：

——增加了“縮略語”（見第4章）；

——增加了ALV?p27抗原免疫膠體金試紙卡檢測方法（見8.1.4.3）；

——更改了抗體檢測結(jié)果對確診的判定標(biāo)準(zhǔn)（見8.4.5，2010年版的4.3.2.3）。

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(dān)識別專利的責(zé)任。

本文件由中華人民共和國農(nóng)業(yè)農(nóng)村部提出。

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

本文件起草單位：山東農(nóng)業(yè)大學(xué)、中華人民共和國拱北海關(guān)技術(shù)中心、華南農(nóng)業(yè)大學(xué)、揚州大學(xué)、

中國農(nóng)業(yè)科學(xué)院哈爾濱獸醫(yī)研究所、靈羽生工（北京）醫(yī)藥科技有限公司、中國農(nóng)業(yè)大學(xué)、普萊柯生物工

程股份有限公司、中國動物疫病預(yù)防控制中心。

本文件主要起草人：趙鵬、崔治中、孫淑紅、楊素、沙才華、廖明、曹偉勝、秦愛建、錢琨、王一新、

高玉龍、陳西釗、蘇敬良、田克恭、吳清民、王傳彬。

本文件及其所代替文件的歷次版本發(fā)布情況為：

——2010年首次發(fā)布為GB/T26436—2010；

——本次為第一次修訂。

Ⅲ

GB/T26436—2025

引言

禽白血?。ˋvianleukosis，AL）是由禽白血病病毒（Avianleukosisvirus，ALV）感染引起的禽類多

種腫瘤性疾病的統(tǒng)稱，主要引起雞的惡性腫瘤和亞臨床感染，導(dǎo)致雞只生長遲緩、產(chǎn)蛋下降和免疫抑

制，是一種能夠通過種蛋垂直傳播的免疫抑制性疾病，我國將其列為三類動物疫病。

ALV屬于反轉(zhuǎn)錄病毒科正反錄病毒亞科甲型反轉(zhuǎn)錄病毒屬，根據(jù)其gp85蛋白的特性目前將

ALV分為A、B、C、D、E、F、G、H、I、J和K共11個亞群。ALV分為內(nèi)源性ALV和外源性ALV兩大

類。內(nèi)源性ALV是指整合進(jìn)宿主細(xì)胞染色體基因組因而能通過染色體垂直傳播的ALV，它可能只是

病毒基因組的不完全片段不產(chǎn)生傳染性病毒粒子，也可能是病毒全基因組因而能產(chǎn)生傳染性病毒粒

子，不過這類病毒通常致病性很低或無致病性，主要是E亞群ALV。外源性ALV是與內(nèi)源性ALV

相對應(yīng)的，指不通過宿主細(xì)胞染色體傳遞給下一代的ALV，包括A、B、C、D、J和K亞群，致病性強的

ALV均屬于外源性病毒。內(nèi)源性ALV的干擾是開展實驗室檢測，特別是進(jìn)行核酸檢測時首要排除

因素。

本病發(fā)生沒有明顯的季節(jié)性或地域性，通常因為飼養(yǎng)種源未凈化ALV的雞苗而發(fā)病，以及在育雛

期與感染雞群混養(yǎng)或者使用存在外源性ALV污染的生物制品而感染發(fā)病。

本文件診斷技術(shù)內(nèi)容包括臨床診斷、病原學(xué)診斷和血清學(xué)診斷方法。本文件的修訂參考了世界動

物衛(wèi)生組織（WOAH）《陸生動物診斷試驗和疫苗手冊》，并結(jié)合了我國相關(guān)技術(shù)研究新成果。

Ⅳ

GB/T26436—2025

禽白血病診斷技術(shù)

1范圍

本文件描述了禽白血病的臨床診斷、樣品采集與處理，以及病毒分離