BACKGROUND: Larval toxocarosis is a zoonosis caused by larvae of Toxocara canis and T. cati, a gastrointestinal nematode of canids and felids, respectively. Diagnosis is usually performed by ELISA IgG using Toxocara excretory-secretory products as an antigen. Due to laboriousness of isolation of the products and subsequent process of standardization of antigenic compounds, routine use of this method is limited and can produce inaccurate diagnostical results. The purpose of this study was to discover new specific antigenic proteins that could be used in routine serological methods of larval toxocarosis. MATERIALS AND METHODS: Toxocara excretory-secretory products were collected and separated by SDS-PAGE. Proteins from the gel were electro-transferred to a membrane and incubated with mouse sera. Antigenic proteins were analyzed using the liquid chromatography-tandem mass spectrometry approach. Selected proteins were prepared in recombinant form and tested with mice and human sera by ELISA and Western blot. RESULTS: A total of four recombinant protein antigens were prepared (rTc-TES-26, rTc-ASA, rTc-PDP, and rTc-ASP). They were analyzed by ELISA and Western blot using mice and human sera. For all sera, three of the four recombinant antigens correlated with Toxocara excretory-secretory products in ELISA analysis. By Western blot, the infection was confirmed in all experimentally infected mice and two out of seven human patients. CONCLUSION: Combination of the presented methods and analyses represents a possible method of effective identification of Toxocara protein antigens for the purpose of routine serodiagnosis.

• Klíčová slova
• Antigen, Diagnostics, Recombinant protein, Toxocara canis, Toxocariasis, Toxocarosis,
• MeSH
• antigeny helmintové MeSH
• ELISA MeSH
• larva MeSH
• lidé MeSH
• myši MeSH
• Toxocara canis * MeSH
• Toxocara MeSH
• toxokaróza * MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• myši MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• antigeny helmintové MeSH

BACKGROUND: Larval toxocarosis (LT), a zoonotic disease transmitted by dogs, cats, and other carnivores, is caused by roundworms of the genus Toxocara. Humans become infected by ingesting embryonated eggs of this parasite. In this study, we present data on the seroprevalence of LT in the Czech Republic collected by the National Reference Laboratory for Tissue Helminthoses in 2012-2016. METHODS: Using enzyme-linked immunosorbent assay, a total of 4428 adults and children with or without clinical symptoms were examined for the presence of IgG antibodies against Toxocara canis excretory-secretory antigens. RESULTS: Of all the persons examined, specific Toxocara antibodies were detected in 160 (3.6%) individuals. There were, however, significant differences between various regions, with seropositivity rates ranging from 1.4 to 7.5%. CONCLUSION: In comparison to studies from 1998 and 2004, our results suggest a decrease in overall Toxocara seroprevalence in the Czech population, whereby the rates are similar to or even lower than rates in some other Central European countries.

• Klíčová slova
• Czech Republic, Seroprevalence, Toxocara canis, Toxocara cati, Toxocarosis, Zoonosis,
• MeSH
• dítě MeSH
• dospělí MeSH
• ELISA MeSH
• imunoglobulin G krev   MeSH
• kojenec MeSH
• larva MeSH
• lidé středního věku MeSH
• lidé MeSH
• mladiství MeSH
• mladý dospělý MeSH
• novorozenec MeSH
• předškolní dítě MeSH
• protilátky helmintové krev   MeSH
• senioři MeSH
• séroepidemiologické studie MeSH
• Toxocara izolace a purifikace   MeSH
• toxokaróza krev   epidemiologie   MeSH
• zoonózy epidemiologie   parazitologie   MeSH
• zvířata MeSH
• Check Tag
• dítě MeSH
• dospělí MeSH
• kojenec MeSH
• lidé středního věku MeSH
• lidé MeSH
• mladiství MeSH
• mladý dospělý MeSH
• mužské pohlaví MeSH
• novorozenec MeSH
• předškolní dítě MeSH
• senioři MeSH
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Geografické názvy
• Česká republika epidemiologie   MeSH
• Názvy látek
• imunoglobulin G MeSH
• protilátky helmintové MeSH

Using a small animal imaging system, migratory activity of Toxocara canis larvae stained by carboxyfluorescein succinimidyl ester (CFSE) was observed post primary infection (PPI) and post reinfection (PR) of BALB/c mice. Each infection was performed with 1,000 larvae per mouse. Primary infections were performed with labeled larvae, while for challenge infections the reinfecting larvae were stained by CFSE. The worm burden in mouse organs was determined during a period from 6 h to 21 days and 4 months PPI and PR. In comparison with primary infections that led to the first larvae appearance in the brain after 60 h, greatly accelerated migration of the parasites administered 3 weeks PPI to the CNS and eyes of challenged mice was noted-in both organs the larvae appeared 6 h PR. In all challenged mice, reinfecting larvae prevailed in the resident parasite population. Preliminary experiments with Toxocara cati larvae also revealed early brain involvement in primarily infected mice. Staining of T. canis larvae by CFSE had no effect on the development of a humoral antibody response against T. canis excretory-secretory antigens. In ELISA, elevated levels of specific IgG and IgG1 were noted on day 14 PPI and the levels of antibodies increased till the end of experiment. Reinfection induced an increase in the levels of both antibodies. In terms of optical density, IgG1 antibodies gave higher values in all sera examined. In ELISA for IgG antibodies, an increase in the avidity index of around 50% was detected 1 month PPI; higher-avidity antibodies were also detected in sera of reinfected animals.

• MeSH
• barvení a značení MeSH
• časové faktory MeSH
• ELISA MeSH
• helmintózy centrálního nervového systému patologie   MeSH
• imunoglobulin G krev   MeSH
• larva patogenita   MeSH
• mozek parazitologie   MeSH
• myši inbrední BALB C MeSH
• myši MeSH
• oči parazitologie   MeSH
• oční nemoci parazitologie   patologie   MeSH
• parazitární zátěž MeSH
• protilátky helmintové krev   MeSH
• Toxocara canis patogenita   MeSH
• toxokaróza parazitologie   patologie   MeSH
• zvířata MeSH
• Check Tag
• myši MeSH
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• imunoglobulin G MeSH
• protilátky helmintové MeSH