The detection of Toxoplasma gondii ME49 infections in BALB/c mice using various techniques.

0 PARASITOLOGY Parasites, hosts and diseases Pub Date : 2023-11-01 Epub Date: 2023-11-28 DOI:10.3347/PHD.23048
Hae-Ji Kang, Jie Mao, Min-Ju Kim, Keon-Woong Yoon, Gi-Deok Eom, Ki-Back Chu, Eun-Kyung Moon, Fu-Shi Quan
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Abstract

Toxoplasma gondii infections are primarily diagnosed by serological assays, whereas molecular and fluorescence-based techniques are garnering attention for their high sensitivity in detecting these infections. Nevertheless, each detection method has its limitations. The toxoplasmosis detection capabilities of most of the currently available methods have not been evaluated under identical experimental conditions. This study aimed to assess the diagnostic potential of enzyme-linked immunosorbent assay (ELISA), real-time polymerase chain reaction (RT-PCR), immunohistochemistry (IHC), and immunofluorescence (IF) in BALB/c mice experimentally infected with various doses of T. gondii ME49. The detection of toxoplasmosis from sera and brain tissues was markedly enhanced in mice subjected to high infection doses (200 and 300 cysts) compared to those subjected to lower doses (10 and 50 cysts) for all the detection methods. Additionally, increased B1 gene expression levels and cyst sizes were observed in the brain tissues of the mice. Importantly, IHC, IF, and ELISA, but not RT-PCR, successfully detected T. gondii infections at the lowest infection dose (10 cysts) in the brain. These findings may prove beneficial while designing experimental methodologies for detecting T. gondii infections in mice.

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采用多种技术检测BALB/c小鼠刚地弓形虫ME49感染。
刚地弓形虫感染主要通过血清学检测来诊断,而分子和荧光技术因其在检测这些感染方面的高灵敏度而受到关注。然而,每种检测方法都有其局限性。目前大多数可用方法的弓形虫病检测能力尚未在相同的实验条件下进行评估。本研究旨在评估酶联免疫吸附试验(ELISA)、实时聚合酶链反应(RT-PCR)、免疫组织化学(IHC)和免疫荧光(IF)对实验感染不同剂量弓形虫ME49的BALB/c小鼠的诊断潜力。在所有检测方法中,高感染剂量(200和300个囊肿)小鼠的血清和脑组织中弓形虫病的检出率明显高于低感染剂量(10和50个囊肿)小鼠。此外,在小鼠脑组织中观察到B1基因表达水平和囊肿大小的增加。重要的是,IHC, IF和ELISA,而不是RT-PCR,成功地检测了大脑中最低感染剂量(10个囊肿)的弓形虫感染。这些发现可能有助于设计检测小鼠弓形虫感染的实验方法。
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