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首页> 外国专利> Indirect elisa kit for serodiagnosis of brucellosis in livestock and humans

Indirect elisa kit for serodiagnosis of brucellosis in livestock and humans

机译：间接ELISA试剂盒用于禽畜和人的布鲁氏菌病血清学诊断

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pTo evolve epidemiology of brucellosis in both human and animal population in the country, a common ELISA kit for screening multiple species of livestock and humans would be desirable. In the present disclosure indirect ELISA has been standardized using smooth lipopolysaccahride antigen from a standard strain of Brucella abortus S99. Antigen dilution of 1:300 was taken as the optimum dilution by checker board titration. Anti-Brucella sLPS hyperimmune sera raised in two brucellosis-negative adult male pigs and sheep and pre-immune serum from same pigs and sheep were used as the positive and negative controls along with convalescent bovine sera respectively. The moderate positive control was prepared by diluting strong positive sera with Brucella negative sera at 1:500 dilutions. ELISA protocol has been standardized using 1:300 diluted smooth lipopolysaccahride antigen from a standard strain of Brucella abortus S99 and protein-G conjugated with horseraddish peroxidase. Sera samples showing percentage positive values of 55 and above were considered Brucella positive. In the protein-G based ELISA test, the sensitivity with respect to detection percentage of positives varied from 100% in pigs and humans followed by 98% in cattle, 96% in goat and sheep and least in buffaloes (92%). Whereas, sensitivity of detection percentage of negatives ranged from 100% in humans followed by 98% in pig and buffaloes, 95% in cattle and least in goat and sheep (92%). The sensitivity of the test was found more than 90% with respect to both positives and negatives in all livestock species and humans. Use of protein-G conjugate (common conjugate) as single reaction agent is advantageous than the use of 4 different conjugates. Also it is easy to maintain the quality of the kit and easy to perform than the use of multiple kits and protocols for different species. The wider application of this protocol will help to obtain the seroepidemiology of brucellosis in both animals and humans. This test will also help to adopt preventive measures like vaccines for farm animals and initiate treatment for humans./p
机译：为了在该国人类和动物种群中发展布鲁氏菌病的流行病学，需要一种用于筛选多种牲畜和人类的通用ELISA试剂盒。在本公开中，已经使用来自标准布鲁氏菌S99的菌株的光滑脂多糖表面抗原对间接ELISA进行了标准化。通过棋盘滴定将抗原稀释度为1：300作为最佳稀释度。在两只布鲁氏菌病阴性的成年雄性猪和绵羊中培养的抗Brucella sLPS超免疫血清以及相同猪和绵羊的免疫前血清分别与恢复期的牛血清作为阳性和阴性对照。中度阳性对照是通过以1：500稀释的布鲁氏菌阴性血清稀释强阳性血清而制备的。 ELISA协议已使用来自标准布鲁氏菌S99菌株和与辣根过氧化物酶偶联的蛋白G的1：300稀释的光滑脂多糖表面抗原进行了标准化。血清百分比阳性值显示为55以上的样品被认为是布鲁氏菌阳性。在基于蛋白质G的ELISA测试中，对阳性检出百分比的敏感性变化范围为猪和人为100％，其次是牛，为98％，山羊和绵羊为96％，水牛（最少为92％）。阴性检测百分比的敏感性范围为：人类为100％，其次是猪和水牛为98％，牛为95％，山羊和绵羊最少（92％）。在所有牲畜物种和人类中，相对于阳性和阴性，该测试的灵敏度均超过90％。蛋白-G缀合物（普通缀合物）作为单一反应剂的使用比使用4种不同的缀合物有利。此外，与针对不同物种使用多个试剂盒和规程相比，维护试剂盒的质量更容易执行。该方案的广泛应用将有助于在动物和人类中获得布鲁氏菌病的血清流行病学。该测试还将有助于采取预防措施，例如为家畜提供疫苗，并开始对人类进行治疗。

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