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[牛牛皮蝇抗体检测ELISA试剂盒的研制。I. 组件的研制]

[Development of the ELISA kit for the detection of Hypoderma bovis antibodies in cattle. I. Development of components].

作者信息

Ziomko I, Cencek T

机构信息

Zakład Parazytologii i Chorób Inwazyjnych, Państwowy Instytut Weterynaryjny 24-100 Puławy, ul. Partyzantów 57.

出版信息

Wiad Parazytol. 2001;47(3):497-503.

Abstract

The aim of these investigations was to prepare the antigen and control sera for the ELISA. Positive control sera were obtained from 163 cows showing clinical symptoms of H. bovis infection and negative control sera from 60 cows free from the infection. The antigen for the ELISA was obtained from H. bovis L1 larvae. Three basic fraction were isolated from whole excreted/secreted antigen by elution from gel and by preparative isoelectrofocusing. Using the Western-Blotting method it was demonstrated that Hy A (22-24 kDa and pI 4-5) had the best antigenic property. As can be seen from these investigations the developed ELISA kit for the detection of H. bovis antibodies in cows is highly specific and sensitive.

摘要

这些研究的目的是制备用于酶联免疫吸附测定(ELISA)的抗原和对照血清。阳性对照血清取自163头表现出牛肝片吸虫感染临床症状的奶牛,阴性对照血清取自60头未感染该疾病的奶牛。ELISA所用的抗原取自牛肝片吸虫L1幼虫。通过凝胶洗脱和制备性等电聚焦从全部排泄/分泌抗原中分离出三个基本组分。采用蛋白质印迹法证明,Hy A(22 - 24 kDa,等电点4 - 5)具有最佳抗原特性。从这些研究中可以看出,所研制的用于检测奶牛牛肝片吸虫抗体的ELISA试剂盒具有高度特异性和敏感性。

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