BackgroundToxocara canis is a nematode that parasitizes dogs, while humans are paratenic hosts. When humans are infected the migrating larvae damage the liver, lungs and even the nervous system. Larva migrans diagnosis is based on immunological techniques; however, the commercial immunodiagnostic kits detect anti-T. canis antibodies which may cross-react with other parasites, mainly nematodes with extra-intestinal migration. Moreover, antibodies do not necessarily reflect an active infection; so detection and quantification of circulating antigens may provide appropriate and timely information for treatment, which prevents irreversible damage. Here we report the standardization of a monoclonal antibody based antigen capture ELISA to diagnose human toxocariasis without cross-reaction.MethodsWe developed anti-T. canis polyclonal antibodies in rabbits and a monoclonal antibody in mouse which did not cross-react with 15 antigens from several parasites. The sandwich ELISA standardization was performed using sera from mice experimentally infected. We tested the method using 29 positive and 58 negative human sera previously typified with a commercial kit, which detects antibodies.ResultsOnly 5.0 μg/mL and 10 μg/mL polyclonal antibodies and monoclonal antibody, respectively, were needed in the sandwich ELISA standardization, detecting since 440 pg/mL larva antigens. Nine out of 29 antibody-positive sera were also positive for antigens and no false positive were found. Taking the antibody kit as the reference standard, the sensibility and specificity of the antigen test were 31% and 100%, respectively.ConclusionsWith these tools we established a detection threshold as low as 440 pg/mL antigen. Monoclonal antibody is specific, and did not cross-react with antigens from other parasites. Detection of circulating antigens helps provide appropriate and timely treatment and prevents irreversible damage.
The erratic migration of nematode larvae is potentially deleterious for humans, especially those produced by Toxocara roundworms. Diagnosis of human toxocariosis: visceral larva migrans depends on antibody detection against excretions-secretions (E/S) from Toxocara larvae by enzyme-linked immunosorbent assay. In the present work, we wish to propose a simple and cheap culture procedure for keeping T. canis female worms alive. We removed living worms from the small intestine of euthanized puppies at the Canine Control Centre "Culhuacan" in the Southern part of Mexico City. To ascertain the utility of the method, we compared the population size of fertilized eggs from both female worms kept in culture (FCM) and those obtained using uterus excision method (UEM). Operationally, a fertilized egg develops a larva after 1 month. Our results show that the culture method was superior at a ratio of 3:1 over UEM. FCM had advantages over UEM, such as (1) low probability of contamination with foreign antigens, (2) greater safety for infective egg manipulation, and (3) gave rise to a higher percentage of fertile eggs.
Introduction:Experiencing or being exposed to violence is called victimization; living it can generate repercussions on health, quality of life and life expectancy. Objectives: To describe victimization and identify factors related to polyvictimization (≥ 4 victimization incidents) in adolescents. Methods: After informed consent and assent were obtained, the ICAST-C and Youth Self-Report validated self-report questionnaires were applied in nine public secondary schools, by means of which demographic data, six forms of victimization and symptoms related to eight mental health problems were investigated. Frequencies of the forms of victimization and polyvictimization were obtained and an ordinal regression was carried out to identify variables related to polyvictimization. Results: The answers of 638 participants were included; 49.37% reported victimization throughout life, 53.37% before previous year and 68.86% during previous year; 47.65% reported polyvictimization, 21.75% before previous year and 17.53% during previous year. The factors related to polyvictimization were depression-introversion, attention deficit, rule-breaking behaviors, bullying and parental separation/divorce. Conclusions: Victimization and polyvictimization were frequent in this sample of adolescents; the factors that were related to polyvictimization included symptoms of mental health problems, bullying and parental divorce/separation.
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