弓形虫是一种主要的食源性人畜共患病原体,可以通过食用小反刍动物的生肉或未煮熟的肉来传播,在其他人中。血清学已被建议作为流行病学指标,如今有几种测试可用。然而,没有与最常用的比较研究。因此,本研究的目的是开发和验证两个内部测试(Westernblot-TgSALUVETWB-和ELISA-TgSALUVETELISA2.0-),并进行比较研究,包括此类测试和四个商业ELISA试剂盒(IDScreen®,PrioCHECK®,Pigtype®和IDEXX)。首先,通过未感染绵羊和感染弓形虫或新孢子虫的绵羊的血清面板确定TgSALUVETWB对免疫显性抗原的特定识别模式。接下来,使用来自天然感染弓形虫的绵羊和山羊的血清,将TgSALUVETWB用作参考,以初步验证TgSALUVETELISA2.0。然后,通过所有测试分析上述绵羊血清面板,并进行TG-ROC分析和一致性测试,与抗N的交叉反应性研究了犬IgG。所有技术对于所有血清面板最初建议的截止值都足够准确(Se和Sp≥94%),除了PrioCHECK®,显示83%的Sp。然而,截止重新调整提高了他们的诊断性能。此外,反-N之间的交叉反应所有测试均检测到犬抗体和弓形虫抗原。因此,我们进行了第二次截止值重新调整,建议使用两个重新调整的截止值,以获得可比较的数据并避免假阳性结果.
Toxoplasma gondii is a major foodborne zoonotic pathogen that can be transmitted through the consumption of raw or undercooked meat of small ruminants, among others. Serology has been suggested as an epidemiological indicator and several tests are available nowadays. However, there is no comparative
study with the most used ones. Therefore, the objective of this
study was to develop and validate two in-house tests (Western blot -TgSALUVET WB- and ELISA -TgSALUVET ELISA 2.0-) and perform a comparative
study including such tests and four commercial ELISA kits (IDScreen®, PrioCHECK®, Pigtype® and IDEXX). First, a specific pattern of recognition of immunodominant antigens by TgSALUVET WB was determined with serum panels of noninfected sheep and sheep infected with T. gondii or
Neospora caninum. Next, TgSALUVET WB was used as a reference to preliminary validate TgSALUVET ELISA 2.0 using sera from sheep and goats naturally infected with T. gondii. Then, the abovementioned sheep serum panels were analyzed by all tests and subjected to TG-ROC analyses and agreement tests, and cross-reactivity with the anti-N. caninum IgGs was studied. All the techniques were accurate enough for the cutoff values initially suggested with all serum panels (Se and Sp ≥ 94%), except for PrioCHECK®, which showed 83% Sp. However, a cutoff readjustment improved their diagnostic performance. Additionally, cross-reactions between anti-N. caninum antibodies and T. gondii antigens were detected with all tests. Thus, a second cutoff readjustment was carried out and the use of both readjusted cutoff values is recommended to obtain comparable data and avoid false-positive results.