Siju Susan Jacob, Pinaki Prasad Sengupta, Samer Shamshad, S. Sudhagar, A.G. S. Chandu, S.S. Patil, S.Madhaba Maharana
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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. 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引用次数: 0
摘要
猪带绦虫囊虫病是一种被忽视的热带病,具有重要的公共卫生意义,特别是在全球发展中国家。猪在猪螺旋体的生命周期中充当中间宿主,并作为人类感染的主要来源。在猪中发现感染对控制人类传播极为重要。本研究对猪囊尾蚴两种低分子量抗原(Ag1和Ag1V1)进行了克隆、测序和分析。经序列确认后,将纯化的PCR产物连接到pET 32b表达载体上,在原核表达系统中表达重组蛋白。用抗囊液抗原的高免疫血清对纯化的重组蛋白进行Western blotting鉴定。此外,本研究旨在利用重组Ag1和Ag1V1抗原建立酶联免疫吸附试验(ELISA)。与囊液抗原相比,使用Ag1检测的敏感性和特异性分别为95.3 %和91.7 %,使用Ag1V1检测的敏感性和特异性分别为93.7 %和97.9% %。基于Ag1 Ag1的ELISA加权Cohen’s kappa为0.726(95 % CI 0.588-0.865),基于Ag1V1的ELISA加权Cohen’s kappa为0.847(95 % CI 0.738-0.955)。因此,该方法可作为猪囊虫病诊断和血清监测的重要工具,从而确定高风险地区,实施战略控制措施。然而,由于这是一项初步研究,通过对感染和未感染的自由漫游猪血清样本的评估进一步确认将揭示更清晰的情况。
Development of an enzyme immunoassay employing low molecular weight recombinant Ag1 and Ag1V1 proteins of cystic fluid for the serodiagnosis of porcine cysticercosis
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.
期刊介绍:
The journal Veterinary Parasitology has an open access mirror journal,Veterinary Parasitology: X, sharing the same aims and scope, editorial team, submission system and rigorous peer review.
This journal is concerned with those aspects of helminthology, protozoology and entomology which are of interest to animal health investigators, veterinary practitioners and others with a special interest in parasitology. Papers of the highest quality dealing with all aspects of disease prevention, pathology, treatment, epidemiology, and control of parasites in all domesticated animals, fall within the scope of the journal. Papers of geographically limited (local) interest which are not of interest to an international audience will not be accepted. Authors who submit papers based on local data will need to indicate why their paper is relevant to a broader readership.
Parasitological studies on laboratory animals fall within the scope of the journal only if they provide a reasonably close model of a disease of domestic animals. Additionally the journal will consider papers relating to wildlife species where they may act as disease reservoirs to domestic animals, or as a zoonotic reservoir. Case studies considered to be unique or of specific interest to the journal, will also be considered on occasions at the Editors'' discretion. Papers dealing exclusively with the taxonomy of parasites do not fall within the scope of the journal.