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      New insights on serodiagnosis of trichinellosis during window period: early diagnostic antigens from Trichinella spiralis intestinal worms

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          Abstract

          The clinical diagnosis of trichinellosis is difficult because its clinical manifestations are nonspecific. Detection of anti- Trichinella IgG by ELISA using T. spiralis muscle larval excretory-secretory (ES) antigens is the most commonly used serological method for diagnosis of trichinellosis, but the main disadvantage is false negativity during the early stage of infection. There is an obvious window period between Trichinella infection and antibody positivity.

          During the intestinal stage of Trichinella infection, the ES antigens of intestinal worms (intestinal infective larvae and adults) are exposed to host’s immune system at the earliest time and elicit the production of specific anti- Trichinella antibodies. Anti- Trichinella IgG antibodies in infected mice were detectable by ELISA with ES antigens of intestinal worms as soon as 8–10 days post infection (dpi), but ELISA with muscle larval ES antigens did not permit detection of infected mice before 12 dpi. Therefore, the new early antigens from T. spiralis intestinal worms should be screened, identified and characterized for early serodiagnosis of trichinellosis.

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          The online version of this article (doi:10.1186/s40249-017-0252-z) contains supplementary material, which is available to authorized users.

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          Survey of Trichinella infections in domestic pigs from northern and eastern Henan, China.

          The aim of this work was to investigate the current situation of Trichinella infections in swine in the cities of Anyang and Shanqiu in the Henan province historically designated as trichinellosis-free. A total of 475 diaphragm muscle samples were collected from 2010 to 2011 and examined by trichinelloscopy and artificial digestion. No Trichinella larvae were detected by trichinelloscopy; however, using the digestion method, 3.79% (18/475) of domestic pigs were deemed positive for Trichinella. Among the 475 pigs examined, 112 from an industrialized pig farm were negative. However. Trichinella larvae were detected in 10% (9/90) of pigs from small pig farms, which was significantly higher than the 3.3% (9/273) of pigs found positive from backyard farms (P<0.05). The larval burdens in infected animals ranged from 0.1 to 1.58 larvae per gram. The larvae were identified by multiplex PCR as Trichinella spiralis. Our study confirms the existence of porcine trichinellosis in northern and eastern parts of Henan. The results will be useful for evaluating the risk of infection for humans. Given this new found data, public health officials should consider implementing strategies to eliminate human transmission. Copyright © 2013 Elsevier B.V. All rights reserved.
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            Early serodiagnosis of trichinellosis by ELISA using excretory–secretory antigens of Trichinella spiralis adult worms

            Background The excretory–secretory (ES) antigens of Trichinella spiralis muscle larvae (ML) are the most commonly used diagnostic antigens for trichinellosis. Their main disadvantage for the detection of anti-Trichinella IgG is false-negative results during the early stage of infection. Additionally, there is an obvious window between clinical symptoms and positive serology. Methods ELISA with adult worm (AW) ES antigens was used to detect anti-Trichinella IgG in the sera of experimentally infected mice and patients with trichinellosis. The sensitivity and specificity were compared with ELISAs with AW crude antigens and ML ES antigens. Results In mice infected with 100 ML, anti-Trichinella IgG were first detected by ELISA with the AW ES antigens, crude antigens and ML ES antigens 8, 12 and 12 days post-infection (dpi), respectively. In mice infected with 500 ML, specific antibodies were first detected by ELISA with the three antigen preparations at 10, 8 and 10 dpi, respectively. The sensitivity of the ELISA with the three antigen preparations for the detection of sera from patients with trichinellosis at 35 dpi was 100 %. However, when the patients’ sera were collected at 19 dpi, the sensitivities of the ELISAs with the three antigen preparations were 100 % (20/20), 100 % (20/20) and 75 % (15/20), respectively (P < 0.05). The specificities of the ELISAs with the three antigen preparations were 98.11, 95.60 and 89.31 %, respectively (P < 0.05). Conclusions The sensitivity and specificity of the T. spiralis AW ES antigens were superior to those of the AW crude antigens and ML ES antigens. Thus, the AW ES antigens might serve as potential antigens for the early and specific serodiagnosis of trichinellosis.
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              Characterization of a Trichinella spiralis 31 kDa protein and its potential application for the serodiagnosis of trichinellosis.

              The Trichinella spiralis 31 kDa protein (Ts31) was screened from the excretory-secretory (ES) proteins of muscle larvae (ML) by immunoproteomics using serum from mice infected with T. spiralis at 18 days post infection (dpi). The aim of this study was to characterize the Ts31 protein and to evaluate the potential of the recombinant Ts31 protein (rTs31) for serodiagnosis of human trichinellosis. Ts31 gene was cloned and rTs31 was produced in an E. coli expression system. An anti-rTs31serum recognized the native protein migrating in a 25-55 kDa range by Western blotting of ML crude or ES antigens. Expression of Ts31 gene was observed at all developmental stages of T. spiralis (adult worms, newborn larvae, pre-encapsulated larvae and ML). An immunolocalization analysis identified Ts31 in the cuticle and stichocytes of the parasite. The sensitivity of rTs31-ELISA and ES antigen ELISA for detecting anti-Trichinella IgG antibodies in sera of patients with trichinellosis was 97.83% (45/46) and 86.78% (39/46), respectively (P>0.05); The specificity of rTs31-ELISA was 99.13% (114/115), which was significantly higher than 85.22% (98/115) of ES antigen ELISA (P<0.01). The rTs31 protein of T. spiralis could be considered as a potential diagnostic antigen for trichinellosis.
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                Author and article information

                Contributors
                wangzq@zzu.edu.cn
                shiyali3071@163.com
                liuruodan2006@126.com
                jpdaisy@126.com
                guan_ml@126.com
                cyingdan@126.com
                cuij@zzu.edu.cn
                Journal
                Infect Dis Poverty
                Infect Dis Poverty
                Infectious Diseases of Poverty
                BioMed Central (London )
                2049-9957
                20 February 2017
                20 February 2017
                2017
                : 6
                Affiliations
                [1 ]ISNI 0000 0001 2189 3846, GRID grid.207374.5, Department of Parasitology, , Medical College, Zhengzhou University, ; Zhengzhou, 450052 China
                [2 ]National Institute of Parasitic Diseases, Chinese Center for Disease Control and Prevention, Key Laboratory of Parasite and Vector Biology, Ministry of Health, Shanghai, 200025 China
                Article
                252
                10.1186/s40249-017-0252-z
                5319148
                28219418
                © The Author(s). 2017

                Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License ( http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver ( http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.

                Funding
                Funded by: FundRef http://dx.doi.org/10.13039/501100001809, National Natural Science Foundation of China;
                Award ID: 81572024
                Award ID: 81672043
                Award Recipient :
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                Opinion
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                © The Author(s) 2017

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