Using all three tests, in combination, raises diagnostic sensitivity

Using all three tests, in combination, raises diagnostic sensitivity. along with goats and pigs possess the highest incidence of cysts in meat, and play a major role as a source of human contamination. Methods In this study, a new commercial ELISA kit (PrioCHECK? Toxoplasma Ab SR, Prionics Schlieren-Zurich, Switzerland) for the detection of anti-antibodies in serum, plasma and meat juice of sheep, was evaluated by comparing it with the indirect fluorescent antibody test (IFAT), indirect haemagglutination test (IHA) and real-time PCR, on samples from experimentally inoculated and naturally uncovered sheep. Results The commercial ELISA detected the infection status in 50% and 100% of sheep orally inoculated with 10,000 oocysts (n?=?6), from two or three weeks post contamination (wpi), respectively, both on serum and plasma samples. Meat juice from all experimentally inoculated sheep collected at slaughter (12 wpi) showed positive ELISA values. In naturally uncovered sheep (n?=?396), the ELISA showed a very good agreement with IFAT (in sheep populations at the farm level or for diagnosis at the slaughterhouse, contributing to the control of this widespread zoonosis. is usually a worldwide-distributed, cyst-forming protozoan parasite that affects warm-blooded animals and humans. Felids, the only known definitive hosts, shed oocysts in their faeces. These oocysts sporulate in the environment and represent the main source of contamination for grazing animals. In sheep, contamination with is considered an important cause of abortion and stillbirth but subclinical infections are also very common. Worldwide, seroprevalences ranging from 4% to 95% have been reported for sheep [1,2]. In these animals, the parasite can persist asymptomatically in the form of bradyzoite-containing tissue BMS-986158 cysts, mainly in the brain and muscle tissue [3]. Among food animals, sheep along with goats and pigs, possess the highest incidence of cysts in meat, and play Rabbit Polyclonal to SHANK2 an important role as a source of contamination for humans [2,4,5]. According to a multicentre caseCcontrol study among pregnant women in Europe, consumption of inadequately cooked or cured meat was the risk factor that most strongly predicted acute contamination with in humans is frequently asymptomatic, it can be life-threatening for congenitally-infected as well as BMS-986158 for immunosuppressed patients [7]. BMS-986158 Therefore, the implementation of new meat safety strategies is an important issue for prevention of transmission to humans [8]. In recent years, in order to improve data collection and to better understand the significance of toxoplasmosis, a scientific panel appointed by the European Food Safety Expert (EFSA) recommended that monitoring of the pre-harvest sector in sheep, goats, pigs and game should occur. They pointed out the need for reference materials and reagents and for well characterized diagnostic methods to be applied to food and animals [9]. So far, diagnostic tools available to detect contamination in sheep include direct methods such as histopathology, immunhistochemistry, polymerase chain reaction (PCR) and bioassays, and indirect (serological) methods, based on the detection of antibodies against the parasite. Serological assessments (e.g. indirect fluorescent antibody test (IFAT), enzyme-linked immune sorbent assay (ELISA), altered agglutination test (MAT), western blot (WB), BMS-986158 latex agglutination test (LAT) and indirect hemagglutination test (IHA) are generally highly sensitive and have been largely used worldwide. Recently, the overall performance of the different serological assessments available for detecting BMS-986158 antibodies against in sheep was compared and discussed [1]. In an effort to facilitate implementation of new, simple reliable assessments we evaluated a new commercial ELISA test kit (PrioCHECK? Toxoplasma Ab SR, Prionics Schlieren-Zurich, Switzerland) for detection of anti-antibodies in serum and meat juice of sheep, by comparing it with the indirect fluorescent antibody test (IFAT), indirect haemagglutination test (IHA) and real-time PCR, using samples derived from naturally uncovered and from experimentally infected sheep. Methods Experimental contamination of sheep with isolate (type II) originally obtained from the faeces of a captive Siberian tiger (seronegative sheep were purchased from a sheep breeder in Kemptthal, Zurich, Switzerland. Six animals were orally inoculated with a suspension made up of 10,000 oocysts of the CZ-Tiger isolate in phosphate buffered saline (PBS). The three remaining sheep received only PBS and served as negative, non-infected controls. All nine animals were clinically monitored over 12 weeks, and blood was taken from the at weeks 0 (inoculation day), 1, 2, 3, 4, 6, 8 and 12 (euthanasia) after inoculation in tubes with and without EDTA. Plasma and serum, respectively, were separated, fractioned, and stored at -20C until use. All six inoculated sheep experienced fever (40.2 up to 41.8C) for 6 or more days between days 3 and 11 post inoculation (dpi), and some of them showed.