Detection of Salmonella Infection in Chickens by an Indirect Enzyme-Linked Immunosorbent Assay Based on Presence of PagC Antibodies in Sera
Detection of Salmonella Infection in Chickens by an Indirect Enzyme-Linked Immunosorbent Assay Based on Presence of PagC Antibodies in Sera
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基于血清中 PagC 抗体的间接酶联免疫吸附测定检测鸡沙门氏菌感染
DOI：
10.1089/fpd.2017.2322
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发表时间：
2017-11-30
影响因子：
2.8
通讯作者：
Fan, Hongjie
中科院分区：
文献类型：
--
作者：
Ma, Zhe;Yang, Xinyi;Fan, Hongjie
人类和动物感染沙门氏菌的结果从持续的无症状携带状态到暂时的轻度胃肠炎或严重的全身感染。快速准确的诊断测试将有助于制定有效预防动物群体感染的策略。目前的测序数据预测，外膜蛋白PagC存在于所有常见的沙门氏菌血清型中，序列相似性超过98%。其他细菌物种中的PagC序列在氨基酸水平上与沙门氏菌的PagC序列的相似性小于65%。我们假设PagC可能具有免疫原性，并且检测该蛋白的抗体可能是沙门氏菌感染的准确指标。将鼠伤寒沙门氏菌CVCC542的pagC基因在大肠杆菌中表达。将纯化的重组PagC蛋白固定在微量滴定板威尔斯孔中。加入经注射感染沙门氏菌或其他非沙门氏菌病原体的SPF鸡的血清，并通过辣根过氧化物酶（HRP）标记的山羊抗鸡抗体检测PagC蛋白的结合。沙门氏菌感染的鸡血清显示出高特异性，与其他细菌感染的鸡血清相比。对87份鸡白痢沙门氏菌抗体阳性血清和93份未感染鸡白痢沙门氏菌抗体阴性血清进行检测，符合率为98.3%。rPagC酶联免疫吸附试验（ELISA）与凝集试验对252份临床鸡血清样品的检测符合率为80.6%。这些结果表明，基于PagC抗体的间接ELISA可以作为一种方便和新的方法，沙门氏菌感染的诊断。
The outcomes of infection of humans and animals with Salmonella range from a persistent asymptomatic carrier state to temporal mild gastroenteritis or severe systemic infection. A rapid and accurate diagnostic test would help formulate strategies for effective prevention of their infections in the animal population. Current sequencing data predict that the outer membrane protein, PagC, is present in all common Salmonella serovars with sequence similarities of more than 98%. PagC sequences in other bacterial species are less than 65% similarity at the amino acid level to those of Salmonella PagC. We hypothesized that PagC could be immunogenic and detection of antibodies to this protein could be an accurate indicator of Salmonella infection. The pagC gene from Salmonella enterica serovar Typhimurium CVCC542 was expressed in Escherichia coli. The purified recombinant PagC protein was immobilized in microtiter plate wells. Sera from SPF chickens infected with Salmonella or other non-Salmonella pathogens by injection were added and binding of PagC protein was detected by the horseradish peroxidase (HRP)-labeled goat anti-chicken antibody. Sera from Salmonella-infected chickens showed high specificity in contrast to the sera from chickens infected with other bacteria. When 87 Salmonella antibody-positive sera from Salmonella Pullorum orally infected SPF chicken and 93 negative sera from uninfected SPF chicken were tested, 98.3% agreement was detected. The rPagC enzyme-linked immunosorbent assay (ELISA) and agglutination had 80.6% agreement in detecting 252 clinical chicken sera samples. These results suggest that PagC antibody-based indirect ELISA can serve as a convenient and novel method for the diagnosis of Salmonella infection.