The use of a synthetic antigen for the serological diagnosis of human trichinellosis (original) (raw)
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Evaluation of Trichinella spiralis larva group 1 antigens for serodiagnosis of human trichinellosis
Journal of Clinical Microbiology, 2004
To identify Trichinella antigens suitable for high-specificity and high-sensitivity serodiagnosis of human trichinellosis, we evaluated assays using four antigens: (i) crude first-stage larval extract (CLE), (ii) O- deglycosylated CLE, (iii) tyvelose-bearing antigens (Trichinella spiralis larva group 1 (TSL-1) antigens) puri- fied by US4 affinity chromatography and coupled directly to enzyme-linked immunosorbent assay (ELISA) plates (pTSL-1 antigens), and (iv) TSL-1 antigens
Comparison of three antigen preparations to detect Trichinellosis in live swine using IgG-ELISA
The Southeast Asian journal of tropical medicine and public health, 2011
A swine infected with Trichinella spiralis is a source of transmission to human through consumption of raw or improperly cooked pork. Detection of larvae is suitable for carcasses, so that pigs in households or farms can be examined serologically for trichinellosis. This study compared antigens, crude (CAg), excretory-secretory (ESAg) and surface (SAg), for their potential use in IgG-ELISA. Serum samples were collected from 5 experimentally infected swine with T. spiralis (pTs), 147 positive cases of 9 other parasitic infections, 12 mixed infections of other parasites, and 35 normal controls. At the same 100% sensitivity, specificity of tests was in a range of 98-77%. ESAg was the best source of antigen with specificity of 98.3% at cut-off value of 0.439. False positives included coccidiasis (1/86) and mixed infections (2/39). For CAg, trichuriasis (2/11), coccidiasis (5/86), and mixed infections (8/39) gave cross-reactions and some of these samples had OD values far above cut-off v...
The estimation of different ELISA procedures for serodiagnosis of human trichinellosis
WiadomoĊci parazytologiczne, 2006
The most important confirmative diagnostic test for trichinellosis is the presence of the muscle larvae in a tissue biopsy but this direct method has a low sensitivity of light and moderate infections. The aim of presented study was to compare the usefulness of the results obtained by three ELISA procedures for Trichinella spp. diagnosis in human outbreaks. All sera (cases and controls) were tested for anti-Trichinella antibodies (immunoglobulin G) using commercially available Novatec KIT and two other ELISA procedures based on excretory-secretory (ES) antigens on Trichinella spiralis muscle larvae. The main differences in ELISA procedures were: the protein concentration in antigen, dilution of human serum samples, conjugate and the time of conjugate incubation. Additional differences were noticed in ES antigen preparation procedures as well as in T. spiralis isolates used in these procedures. Serum samples were obtained from 22 symptomatical patients from Poznafi region (West Polan...
Diagnosis of human trichinellosis : pitfalls in the use of a unique immunoserological technique
Parasite, 2001
Serum samples belonging to three outbreaks in Argentina (47 patients) taken at different times post-ingestion were analysed employing IIF and ELISA simultaneously. Results show that: a) the number of patients diagnosed by a unique technique, especially by ELISA (31 patients), was lower than the one obtained by the simultaneous use of both assays (38 patients); b) four patients out of the seven diagnosed by a unique technique were negative by the other assay over the period of time evaluated. Therefore, It can be concluded that the use of a sole immunoserological technique can not only lead to the delay in the detection but also to the misdiagnosis of this parasitic Infection.
Veterinary Parasitology, 2013
An immunochromatographic strip method, developed with the excretory-secretory antigens from muscle larvae (ML) of Trichinella spiralis labeled with colloidal gold, was used for the detection of anti-Trichinella antibodies in serum of experimentally-infected swine. Sera from swine infected with 200, 2000 and 20,000 infective ML were collected at different days post infection (dpi) and used to evaluate the method. The strip method was shown able to detect anti-Trichinella antibodies by 35 dpi, 28 dpi and 21 dpi for the three different infection doses, respectively, and closely correlated with the results of an ELISA test. The strip method is rapid and easy to perform and is suggested as an acceptable alternative for clinical laboratories lacking specialized equipment, and for field diagnosis of trichinellosis.
Evaluation of excretory-secretory antigens for the serodiagnosis of swine trichinellosis
Veterinary Parasitology, 1988
Gamble, H.R., Rapid, D., Marinculid, A. and Murrell, K.D., 1988. Evaluation of excretory-secretory antigens for the serodiagnosis of swine trichinellosis. Vet. Parasitol., 30: 131-137. Groups of hog sera from endemic and non-endemic areas for swine trichinellosis in Yugoslavia were tested by ELISA using excretory-secretory (ES) antigens collected from T. spiralis muscle larvae maintained in vitro for 24, 48 or 72 h. The 24-h ES had the highest level of specificity for T. spiralis infection. Antigen preparations recovered after 48 or 72 h yielded an increasing rate of false-positive reactions. Additional antigens occured in the 48-and 72-h ES preparations as determined by gel electrophoresis and monoclonal antibody binding. The occurrence of false-negative reactions was directly correlated with T. spiralis worm burdens. Hogs with muscle larvae densities > 10 larvae per gram were all positive by ELISA. Among 17 hogs with < 10 larvae per gram, only one hog was negative by ELISA with 24-h ES antigen; the false-negative rate was higher with 48-and 72-h ES. These results show that ES antigen produced during the first 24 h of in vitro cultivation is highly specific for the immunodiagnosis of swine trichinellosis. spiralis muscle larvae in serum-free tissue culture media .
Serological tools for detection of Trichinella infection in animals and humans
One health (Amsterdam, Netherlands), 2016
Trichinellosis is a serious foodborne zoonotic disease. It is an important threat to public health in both developing and developed countries. Human infections are strongly associated with consuming undercooked meat containing infective Trichinella larvae. The development of serological tools has enabled seroepidemiological studies and contributed to our knowledge on the importance of this parasite. Serological tests can also help the diagnosis of parasite infections in humans and the surveillance of animals. Generally speaking, serological techniques include detection methods for specific antibodies and for circulating parasite antigens in the serum or tissue fluids. Here, we present a comprehensive review of various methods used in the detection of antibodies against Trichinella and circulating parasite antigens in animals and humans.
Veterinary Parasitology, 2014
A clone, designated L20h-Ts3, was selected by immunoscreening of cDNA libraries of Trichinella spiralis worms collected 14 h, 20 h and 48 h post-infection (p.i.) from mice intestines. L20h-Ts3 encodes the full-length of a conserved hypothetical protein of 13.1 kDa involving putative interaction with the immune system. PCR analysis showed that L20h-Ts3 mRNA is constitutively expressed throughout T. spiralis life cycle and not restricted to intestinal stages. The L20h-Ts3 fusion protein was obtained in an Escherichia coli expression system and purified by Ni-affinity chromatography before inoculation into mice in order to produce polyclonal antibodies. Then, immunohistochemical study and Western blot analysis revealed its presence within the stichosome of T. spiralis and in excretory/secretory products strengthening a putative fundamental role for the parasite's survival such as host tissue invasion or modification of the host muscular cell phenotype. L20h-Ts3 fusion protein was recognized in Western blot as soon as 15-20 days p.i. by sera from pigs experimentally infected with 20,000 muscle larvae (ML) of T. spiralis. Thus, an indirect L20h-Ts3 ELISA was designed and evaluated using sera from experimentally infected pigs by comparison with the only ELISA currently available for trichinellosis purposes. A gain of precocity from 7 up to 14 days and detection up to 25 weeks p.i. was possible with the L20h-Ts3 ELISA offering a large window for trichinellosis detection. The L20h-Ts3 ELISA was less effective in the case of low infections in pigs. Nevertheless, these results show that the L20h-Ts3 ELISA has a real interest due to its precocity and stability of detection in time. The association of the L20h-Ts3 fusion protein with other antigenic proteins identified previously could appreciably improve the serological test and facilitate its standardization.
Development and Evaluation of a Western Blot Kit for Diagnosis of Human Trichinellosis
Clinical and Vaccine Immunology, 2003
We evaluated industrially prepared Western blot strips designed to avoid the cross-reactions observed with indirect immunofluorescence and enzyme-linked immunosorbent assays used for the serodiagnosis of trichinellosis. The antigen preparations were crude extracts of Trichinella spiralis. The Western blot profile characteristic of trichinellosis was characterized by comparing 60 sera from patients infected by Trichinella to 11 sera from healthy subjects, 51 sera from patients with other proven parasitic diseases (cysticercosis, schistosomiasis, strongyloidosis, fascioliasis, toxocariasis, liver amebiasis, anisakiasis, filariasis, toxoplasmosis, hydatidosis, or malaria), and 23 sera from patients with autoantibodies. Specific 43-to 44-kDa and 64-kDa bands were obtained with all of the sera from 51 patients with acute trichinellosis, in 4 out of 9 patients at the early stages of the disease, and in only 1 control patient, who had suspected anisakiasis and in whom trichinellosis could not be ruled out by muscle biopsy.