CN1299972A - 犬粪抗原检测酶联免疫吸附试剂盒 - Google Patents

犬粪抗原检测酶联免疫吸附试剂盒 Download PDF

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CN1299972A
CN1299972A CN 00128713 CN00128713A CN1299972A CN 1299972 A CN1299972 A CN 1299972A CN 00128713 CN00128713 CN 00128713 CN 00128713 A CN00128713 A CN 00128713A CN 1299972 A CN1299972 A CN 1299972A
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CN1161610C (zh
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焦伟
柴君杰
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XINJIANG UYGUR AUTONOMOUS REGION INST OF ENDEMIC DISEASE
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XINJIANG UYGUR AUTONOMOUS REGION INST OF ENDEMIC DISEASE
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Abstract

本发明的犬粪抗原检测酶联免疫吸附试剂盒,涉及防治牲畜寄生虫病传染源的治理领域,特别是防治牲畜包虫病的检测器具,本发明提出一种犬粪抗原检测酶联免疫吸附试剂盒,该产品是通过应用ELISA试验检测犬粪中棘球绦虫成虫虫体抗原和排泄分泌抗原的微量诊断方法,用于检测感染棘球绦虫的家犬粪抗原具有足够高的灵敏性和特异性,诊断犬棘球绦虫感染取得成功,并经多次改进,现已制成具体有效的检测成品,即犬粪抗原检测酶联免疫吸附试剂盒,为包虫病的预防和检测工作提供了准确简便有效的检测器具。

Description

犬粪抗原检测酶联免疫吸附试剂盒
本发明的犬粪抗原检测酶联免疫吸附试剂盒，涉及防治牲畜寄生虫病传染源的治理领域，特别是防治牲畜包虫病的检测器具。
感染棘球绦虫的家犬是包虫病的传染源，确定家犬是否感染棘球绦虫是包虫病预防措施的重要组成部分，也是进行包虫病检测和预防效果考核的基本指标，国际传统的方法是用槟榔碱给犬灌服法，但这种方法工作量大，费时、费力、费人、风险较多，且有20％的犬对槟榔碱无反应，即使在反应犬中也不能完全驱出棘球绦虫，因此，国外早就开始寻求新的简便有效的方法，利用免疫学方法检测犬粪中的棘球绦虫抗原是最受重视的方法，英、瑞士的学者取得过较好的成绩，但尚未有具体有效的检测成品器具试制成功。
本发明的目的是提出一种犬粪抗原检测酶联免疫吸附试剂盒。该产品是通过应用ELISA试验检测犬粪中棘球绦虫成虫虫体抗原和排泄分泌抗原的微量诊断方法，用于诊断犬是否感染棘球绦虫？取得了成功，并经多次改进，现已制成具体有效的检测成品，即犬粪抗原检测酶联免疫吸附试剂盒，为包虫病的预防和检测工作提供了准确简便的检测器具。
应用ELISA试验检测犬粪中棘球绦虫成虫虫体抗原和排泄分泌抗原的微量诊断方法的主要技术方案：
1．棘球绦虫成虫虫体→提取抗原→免疫家兔→从血清中提取IgG→亲和层析法纯化IgG作为诊断抗体Ⅰ。
2．棘球绦虫成虫→人工培养→收取培养液→提取排泄分泌抗原→免疫家兔→从血清中提取IgG→亲和层析法纯化IgG作为诊断抗体Ⅱ。
3．将诊断抗体Ⅰ或诊断抗体Ⅱ用辣根过氧化物酶或碱性磷酸酶标记做为诊断的第二抗体，(指示抗体或标记抗体)
4．用抗体Ⅰ或抗体Ⅱ包被酶标板，加待检犬粪标本，加标记抗体，加底物显示试验结果==双抗体夹心酶联免疫吸附试验。
应用ELISA试验检测犬粪中棘球绦虫成虫虫体抗原和排泄分泌抗原的微量诊断方法的实施方案：
1．试剂盒成份：a．小牛血清      3ml×1瓶b．微孔反应板    8孔×6或12孔×4c．酶结合抗体    3.2 ml×1瓶d．阳性对照      1ml×1瓶e．阴性对照    1ml×1瓶f．洗涤液()    30ml×1瓶g．显色剂A     4ml×1瓶h．显色剂B     4ml×1瓶1．终止液      3.5ml×1瓶m．封板片       3片n．犬粪标本保存液
2．实施操作方法：a．取待检犬粪2g放入含2ml保存液的试管中，充分混合后，离心2500rpm10分钟。b．孔中加入小牛血清和待检犬粪标本各一滴，设阴、阳性对照各2孔，每孔加入阴性对照或阳性对照各2滴，在37℃温度下封板1小时。c．洗板甩去孔内液体，注入洗液，放10秒钟，甩干，重复4次，拍干。d．每孔加酶结合抗体1滴，在37℃温度下封板1小时。e．洗板同前。f．加显色剂A、显色剂B各1滴，在37℃温度下停放20分钟，每孔加终止液1滴。g．读取450nmOD值。
应用ELISA试验检测犬粪中棘球绦虫成虫虫体抗原和排泄分泌抗原的微量诊断方法，用于检测感染棘球绦虫的家犬粪抗原具有足够的灵敏性和特异性，替代了用槟榔碱给犬灌服的方法，节约了人力、时间、提高了诊断水平，克服了操作风险。根据应用ELISA试验，检测犬粪中棘球绦虫成虫虫体抗原和排泄分泌抗原的微量诊断方法研制的，本发明的犬粪抗原检测酶联免疫吸附试剂盒能在2～5℃的温度条件下，保存一年，性质不变，能够适用于农牧区基层开展人、畜包虫病的防、治工作。

Claims (2)

1．一种犬粪抗原检测酶联免疫吸附试剂盒，其特征是盒内的成份是装有牛血清3ml×1瓶；微孔反应板8孔×6或12孔×4；酶结合抗体3.2ml×1瓶；阳性对照1ml×1瓶；阴性对照1ml×1瓶；洗涤液(用前，用蒸馏水稀释25倍)30ml×1瓶；显色剂A4ml×1瓶；显色剂B4ml×1瓶；终止液3.5ml×1瓶；封板片3片；犬粪标本保存液1瓶。
2．如权利要求1的一种犬粪抗原检测酶联免疫吸附试剂盒，其特征是所述的犬粪抗原检测酶联免疫吸附试剂盒的实施操作方法是：取待检犬粪2g放入含2ml保存液的试管中，充分混合后，离心2500rpm10分钟；孔中加入小牛血清和待检犬粪标本各一滴，设阴、阳性对照各2孔，每孔加入阴性对照或阳性对照各2滴，在37℃温度下封板1小时；洗板甩去孔内液体，注入洗液，放10秒钟，甩干，重复4次，拍干；每孔加酶结合抗体1滴，在37℃温度下封板1小时；洗板同前；加显色剂A、显色剂B各1滴，在37℃温度下停放20分钟，每孔加终止液1滴；读取450nmOD值。
CNB001287133A 2000-09-08 2000-09-08 犬粪抗原检测酶联免疫吸附试剂盒 Expired - Fee Related CN1161610C (zh)

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Cited By (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN101949926A (zh) * 2010-08-04 2011-01-19 浙江大学 人体包虫病胶体金免疫层析试尿液快速诊断试纸卡
CN101948521A (zh) * 2010-09-17 2011-01-19 中国疾病预防控制中心寄生虫病预防控制所 诊断细粒棘球蚴病的重组抗原蛋白、其制备方法和用途
CN102363808A (zh) * 2011-11-07 2012-02-29 中国农业科学院兰州兽医研究所 从犬粪便中检测细粒棘球绦虫病原的试剂盒
CN103792230A (zh) * 2014-01-13 2014-05-14 东北农业大学 一种奶牛酮病诊断试剂盒及其检测方法
CN107402298A (zh) * 2016-05-18 2017-11-28 上海新吉而生物科技有限公司 一种犬粪包虫病抗原快速检测试剂盒及其使用方法
CN112755049A (zh) * 2021-02-05 2021-05-07 哈密市动物疫病预防控制中心 用于提高犬绦虫病粪便检测灵敏度的泻剂配方及制备方法

Cited By (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN101949926A (zh) * 2010-08-04 2011-01-19 浙江大学 人体包虫病胶体金免疫层析试尿液快速诊断试纸卡
CN101948521A (zh) * 2010-09-17 2011-01-19 中国疾病预防控制中心寄生虫病预防控制所 诊断细粒棘球蚴病的重组抗原蛋白、其制备方法和用途
CN101948521B (zh) * 2010-09-17 2012-08-29 中国疾病预防控制中心寄生虫病预防控制所 诊断细粒棘球蚴病的重组抗原蛋白、其制备方法和用途
CN102363808A (zh) * 2011-11-07 2012-02-29 中国农业科学院兰州兽医研究所 从犬粪便中检测细粒棘球绦虫病原的试剂盒
CN103792230A (zh) * 2014-01-13 2014-05-14 东北农业大学 一种奶牛酮病诊断试剂盒及其检测方法
CN107402298A (zh) * 2016-05-18 2017-11-28 上海新吉而生物科技有限公司 一种犬粪包虫病抗原快速检测试剂盒及其使用方法
CN112755049A (zh) * 2021-02-05 2021-05-07 哈密市动物疫病预防控制中心 用于提高犬绦虫病粪便检测灵敏度的泻剂配方及制备方法

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