Evaluation of ELISA and Western Blot Analysis using three antigens to detect anti-Trichinella IgG in horses.
Abstract: We assessed a serological method for detecting Trichinella infection in horses, specifically, an ELISA using three antigens to detect anti-Trichinella IgG (i.e. a synthetic tyvelose glycan-BSA (stg-BSA) antigen, an excretory/secretory (ES) antigen, and a crude worm extract (CWE) antigen). Serum samples were collected from 2502 horses (433 live horses from Romania and 2069 horses slaughtered in Italy and originating from Italy, Poland, Romania, and Serbia). Serum samples were also taken from horses experimentally infected with different doses of T. spiralis and T. murrelli larvae, as controls. The cut-off value of ELISA was determined on serum samples from 330 horses from Trichinella-free regions of Italy, which were also examined by artificial digestion of preferential-muscle samples. In the experimentally infected horses, the stg-BSA and ES antigens were less sensitive than the CWE antigen. Trichinella spiralis showed a higher immunogenicity than T. murrelli, and the IgG immunoresponse was dose-dependent. The kinetics of anti-Trichinella IgG were similar among all experimentally infected horses. No circulating antibodies were detected 4-5 months after experimental infection, although these horses still harbored infective larvae. Depending on the antigen used, for 4-7 of the 330 horses from Trichinella-free areas, the optical density (OD) of the serum sample was higher than the cut-off value, yet these samples were negative when subjected to Western Blot. Similar results were obtained for the 1739 horses slaughtered in Italy (originating from Italy, Poland, Romania, and Serbia) and the 433 live Romanian horses. Of the 4 horses with muscle larvae, only one was positive by ELISA and Western Blot. Because the anti-Trichinella IgG remain circulating for only a short period of time, whereas the larvae remain infective for longer periods, serology cannot be used for either diagnosing Trichinella infection in horses or estimating the prevalence of infection. Artificial digestion of at least 5 g of preferential-muscle tissue continues to be the method of choice at the slaughterhouse for preventing equine-borne trichinellosis in humans.
Publication Date: 2002-09-05 PubMed ID: 12208044DOI: 10.1016/s0304-4017(02)00185-1Google Scholar: Lookup
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- Evaluation Study
- Journal Article
- Research Support
- Non-U.S. Gov't
Summary
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The research is about the evaluation of Enzyme-Linked Immunosorbent Assay (ELISA) and Western Blot Analysis techniques using three different antigens to detect anti-Trichinella IgG in horses. The study concludes that these methods are inadequate for diagnosing Trichinella infection in horses or estimating prevalence due to short duration of detectable antibodies.
Objective and Methodology
- The researchers aimed to evaluate the effectiveness of a serological method – ELISA, using three antigens (synthetic tyvelose glycan-BSA, an excretory/secretory antigen, and a crude worm extract antigen) to detect anti-Trichinella IgG in horses. The Trichinella infection can lead to the disease called trichinellosis, which is harmful to both horses and humans.
- Serum samples were taken from 2502 horses from various countries and also from horses experimentally infected with Trichinella spiralis and T. murrelli larvae to serve as controls.
- The cut-off value of ELISA was determined using serum samples from 330 horses living in Trichinella-free regions in Italy. These samples were also examined using artificial digestion of muscle samples.
Findings and Analysis
- In the experimentally infected horses, it was found that the synthetic tyvelose glycan-BSA and excretory/secretory antigens were less sensitive than the crude worm extract antigen. The immunogenicity of T. spiralis was higher than that of T. murrelli.
- The IgG immunoresponse was found to be dose-dependent, with the kinetics of anti-Trichinella IgG being similar among all experimentally infected horses. Yet, no circulating antibodies were detected 4-5 months after infection, despite the horses still harboring infective larvae.
- Depending on the antigen used, a few horses from Trichinella-free areas had a higher than cut-off value optical density (OD) of the serum sample, yet they were negative when tested with a Western Blot.
Conclusion
- The serological tests showed that the duration of detectable anti-Trichinella IgG antibodies is short, whereas the larval infectivity remains for longer periods. This means that serological methods like ELISA and Western Blot cannot be reliably used for diagnosing Trichinella infections in horses or for estimating the infection prevalence.
- The study reiterated that artificial digestion of a muscle tissue sample continues to be the preferred detection method at the slaughterhouse for preventing equine-borne trichinellosis in humans.
Cite This Article
APA
Pozio E, Sofronic-Milosavljevic L, Gomez Morales MA, Boireau P, Nöckler K.
(2002).
Evaluation of ELISA and Western Blot Analysis using three antigens to detect anti-Trichinella IgG in horses.
Vet Parasitol, 108(2), 163-178.
https://doi.org/10.1016/s0304-4017(02)00185-1 Publication
Researcher Affiliations
- Laboratory of Parasitology, Istituto Superiore di Sanità, Viale Regina Elena 299, 00161, Rome, Italy. pozio@iss.it
MeSH Terms
- Animals
- Antibodies, Protozoan / blood
- Antigens, Protozoan / immunology
- Blotting, Western / methods
- Blotting, Western / veterinary
- Enzyme-Linked Immunosorbent Assay / methods
- Enzyme-Linked Immunosorbent Assay / veterinary
- Europe
- Horse Diseases / blood
- Horse Diseases / diagnosis
- Horse Diseases / parasitology
- Horses
- Immunoglobulin G / blood
- Immunoglobulin G / immunology
- Meat / parasitology
- Trichinella / isolation & purification
- Trichinellosis / blood
- Trichinellosis / diagnosis
- Trichinellosis / parasitology
- Trichinellosis / veterinary
Citations
This article has been cited 11 times.- Elghandour MMMY, Maggiolino A, Vázquez-Mendoza P, Alvarado-Ramírez ER, Cedillo-Monroy J, De Palo P, Salem AZM. Moringa oleifera as a Natural Alternative for the Control of Gastrointestinal Parasites in Equines: A Review. Plants (Basel) 2023 May 8;12(9).
- Gómez-Morales MÁ, Cherchi S, Ludovisi A. Serological testing for Trichinella infection in animals and man: Current status and opportunities for advancements. Food Waterborne Parasitol 2022 Jun;27:e00165.
- Bruschi F, Gómez-Morales MA, Hill DE. International Commission on Trichinellosis: Recommendations on the use of serological tests for the detection of Trichinella infection in animals and humans. Food Waterborne Parasitol 2019 Mar;14:e00032.
- Stachyra A, Zawistowska-Deniziak A, Basałaj K, Grzelak S, Gondek M, Bień-Kalinowska J. The Immunological Properties of Recombinant Multi-Cystatin-Like Domain Protein From Trichinella Britovi Produced in Yeast. Front Immunol 2019;10:2420.
- Xiao Y, Xu S, Zhao S, Liu K, Lu Z, Hou Z. Protective effects of selenium against zearalenone-induced apoptosis in chicken spleen lymphocyte via an endoplasmic reticulum stress signaling pathway. Cell Stress Chaperones 2019 Jan;24(1):77-89.
- Chu KB, Kim SS, Lee SH, Lee DH, Kim AR, Quan FS. Immune Correlates of Resistance to Trichinella spiralis Reinfection in Mice. Korean J Parasitol 2016 Oct;54(5):637-643.
- Franssen FF, Fonville M, Takumi K, Vallée I, Grasset A, Koedam MA, Wester PW, Boireau P, van der Giessen JW. Antibody response against Trichinella spiralis in experimentally infected rats is dose dependent. Vet Res 2011 Nov 30;42(1):113.
- Gottstein B, Pozio E, Nöckler K. Epidemiology, diagnosis, treatment, and control of trichinellosis. Clin Microbiol Rev 2009 Jan;22(1):127-45, Table of Contents.
- Frey CF, Schuppers ME, Nöckler K, Marinculić A, Pozio E, Kihm U, Gottstein B. Validation of a Western Blot for the detection of anti-Trichinella spp. antibodies in domestic pigs. Parasitol Res 2009 Jun;104(6):1269-77.
- Korínková K, Pavlícková Z, Kovarcík K, Koudela B. Distribution of muscle larvae and antibody dynamics in goats experimentally infected with Trichinella spiralis. Parasitol Res 2006 Nov;99(6):643-7.
- Escalante M, Romarís F, Rodríguez M, Rodríguez E, Leiro J, Gárate MT, Ubeira FM. Evaluation of Trichinella spiralis larva group 1 antigens for serodiagnosis of human trichinellosis. J Clin Microbiol 2004 Sep;42(9):4060-6.
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