The detection of antibody to Bacillus anthracis recombinant PA in vaccinated animal serum

Javkhlan Navaan, Tuvshinzaya Zorigt, Lkham Baasansuren, Narantuya Ayushjav, Enkhtuya Jargalsaikhan
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Abstract

Anthrax is a worldwide zoonosis in animals and human. In Mongolia, the confirmed case of anthrax outbreak is reported every year over the past decade. The prevention and control measure of animal anthrax is vaccination using spore of attenuated Sterne strain, but horse does not get vaccinated in Mongolia. In this study, we constructed the recombinant plasmid for over expression of anthrax protective antigen (PA)/GST fusion protein in pGEX-6P-1 vector and purified the recombinant PA (r-PA) using glutathione Sepharose column under native and denaturing conditions. Since both forms of r-PA were recognized by specific antibody against PA, ELISA system to detect antibody titer in vaccinated bovine serum was constructed. Total of 890 vaccinated cattle serum were collected from 178 cattle at 0, 3, 5, 8 and 12 months’ post vaccination. As negative control, 200 cattle serum from Umnugovi aimag were selected which does not have anthrax foci. All serum was tested by rPA indirect ELISA and, antibody to PA were detected in vaccinated cattle serum but were not detected in negative serum. Therefore, rPA should be used in as monitoring of the anthrax vaccination.
免疫动物血清中重组炭疽芽孢杆菌抗体的检测
炭疽热是一种世界性的人畜共患病。在过去十年中,蒙古每年都报告炭疽热爆发的确诊病例。动物炭疽的预防和控制措施是利用斯特恩减毒菌株孢子接种疫苗,但蒙古没有对马接种疫苗。本研究构建了炭疽保护抗原(PA)/GST融合蛋白在pGEX-6P-1载体上过表达的重组质粒,并在天然和变性条件下利用谷胱甘肽Sepharose柱纯化重组PA (r-PA)。由于两种形式的r-PA均能被特异性抗体识别,因此建立了ELISA检测接种牛血清中抗体滴度的方法。在接种后0、3、5、8和12个月共从178头牛收集了890份接种牛血清。选取未发生炭疽疫源地的乌努戈维牛血清200头作为阴性对照。所有血清均采用rPA间接ELISA检测,接种牛血清中检测到PA抗体,阴性血清中未检测到PA抗体。因此,rPA可用于炭疽疫苗接种的监测。
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