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利用囊液的低分子量重组Ag1和Ag1V1蛋白开发用于猪囊尾蚴病血清诊断的酶免疫测定法。

Development of an enzyme immunoassay employing low molecular weight recombinant Ag1 and Ag1V1 proteins of cystic fluid for the serodiagnosis of porcine cysticercosis.

作者信息

Jacob Siju Susan, Sengupta Pinaki Prasad, Shamshad Samer, Sudhagar S, Chandu A G S, Patil S S, Maharana S Madhaba

机构信息

ICAR National Institute of Veterinary Epidemiology and Disease Informatics, Ramagondanahalli, Yelahanka, Bengaluru 560064, India.

ICAR National Institute of Veterinary Epidemiology and Disease Informatics, Ramagondanahalli, Yelahanka, Bengaluru 560064, India.

出版信息

Vet Parasitol. 2025 Aug;338:110532. doi: 10.1016/j.vetpar.2025.110532. Epub 2025 Jun 20.

DOI:10.1016/j.vetpar.2025.110532
PMID:40544782
Abstract

Taenia solium cysticercosis is one of the neglected tropical diseases with public health importance especially in developing countries across the globe. Pigs serve as the intermediate host in the life cycle of T.solium and acts as a major source of infection to humans. Detecting the infection in pigs is extremely important in controlling the transmission in humans. In this study, the two low molecular weight antigens (Ag1 and Ag1V1) of T. solium cysticerci were cloned, sequenced and analyzed. After sequence confirmation, the purified PCR products were ligated into pET 32b expression vector and the recombinant proteins were expressed in prokaryotic expression system. The purified recombinant proteins were characterized by Western blotting using the hyperimmune sera raised against cyst fluid antigens. Further, the present study was aimed to develop enzyme linked immuno sorbent assays (ELISA) using recombinant Ag1 and Ag1V1 antigens. The sensitivity and specificity of the assay using Ag1 remained 95.3 % and 91.7 %, respectively and using Ag1V1 they were 93.7 % and 97.9 %, respectively, when compared with cystic fluid antigen. The weighted Cohen's kappa was recorded as 0.726 (95 % CI 0.588-0.865) for Ag1 Ag1 based ELISA and 0.847 (95 % CI 0.738-0.955) for the Ag1V1 based one. Hence, the developed assay can be exploited as a valuable tool in the diagnosis and sero-surveillance of porcine cysticercosis and thereby the high-risk areas can be identified to implement strategic control measures. However, as it is a preliminary study, further confirmation by assessment of infected and non-infected free roaming pig serum samples will reveal the clearer picture.

摘要

猪带绦虫囊尾蚴病是一种被忽视的热带疾病,对全球尤其是发展中国家的公共卫生具有重要意义。猪是猪带绦虫生命周期中的中间宿主,也是人类感染的主要来源。检测猪的感染情况对于控制人类传播极为重要。在本研究中,对猪带绦虫囊尾蚴的两种低分子量抗原(Ag1和Ag1V1)进行了克隆、测序和分析。序列确认后,将纯化的PCR产物连接到pET 32b表达载体中,并在原核表达系统中表达重组蛋白。使用针对囊液抗原产生的超免疫血清通过蛋白质印迹法对纯化的重组蛋白进行表征。此外,本研究旨在开发使用重组Ag1和Ag1V1抗原的酶联免疫吸附测定(ELISA)。与囊液抗原相比,使用Ag1的测定的敏感性和特异性分别为95.3%和91.7%,使用Ag1V1的测定的敏感性和特异性分别为93.7%和97.9%。基于Ag1的ELISA的加权科恩kappa系数记录为0.726(95%CI 0.588 - 0.865),基于Ag1V1的ELISA的加权科恩kappa系数记录为0.847(95%CI 0.738 - 0.955)。因此,所开发的测定方法可作为猪囊尾蚴病诊断和血清学监测的有价值工具,从而可以确定高风险地区以实施战略控制措施。然而,由于这是一项初步研究,通过评估感染和未感染的自由放养猪血清样本进行进一步确认将揭示更清晰的情况。

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