应用重组弓形虫过氧化物酶蛋白斑点免疫金银染色法检测孕妇弓形虫感染
Evaluation of toxoplasmosis in pregnant women using dot-immunogold-silver staining with recombinant Toxoplasma gondii peroxiredoxin protein.
机构信息
Jiangsu Key Laboratory of Immunity and Metabolism, Department of Pathogen Biology and Immunology, Xuzhou Medical University, Xuzhou, 221004, China.
出版信息
BMC Infect Dis. 2020 Sep 22;20(1):694. doi: 10.1186/s12879-020-05414-8.
BACKGROUND
Toxoplasma gondii infection endangers human health and affects animal husbandry. Serological detection is the main method used for epidemiological investigations and diagnosis of toxoplasmosis. The key to effective diagnosis of toxoplasmosis is the use of a standardized antigen and a specific and sensitive detection method. Peroxiredoxin is an antigenic protein and vaccine candidate antigen of T. gondii that has not yet been exploited for diagnostic application.
METHODS
In this study, recombinant T. gondii peroxiredoxin protein (rTgPrx) was prepared and used in dot-immunogold-silver staining (Dot-IGSS) to detect IgG antibodies in serum from mice and pregnant women. The rTgPrx-Dot-IGSS method was established and optimized using mouse serum. Furthermore, serum samples from pregnant women were analyzed by rTgPrx-Dot-IGSS.
RESULTS
Forty serum samples from mice infected with T. gondii and twenty negative serum samples were analyzed. The sensitivity and specificity of rTgPrx-Dot-IGSS were 97.5 and 100%, respectively, equivalent to those of a commercial ELISA kit for anti-Toxoplasma IgG antibody. Furthermore, 540 serum samples from pregnant women were screened with a commercial ELISA kit. Eighty-three positive and 60 negative serum samples were analyzed by rTgPrx-Dot-IGSS. The positive rate was 95.18%, comparable to that obtained with the commercial ELISA kit.
CONCLUSIONS
The Dot-IGSS method with rTgPrx as an antigen might be useful for diagnosing T. gondii infection in individuals.
背景
弓形虫感染危害人类健康，影响畜牧业。血清学检测是进行弓形虫病流行病学调查和诊断的主要方法。有效诊断弓形虫病的关键是使用标准化抗原和特异性、敏感性检测方法。过氧化物还原酶是弓形虫的一种抗原蛋白和疫苗候选抗原，尚未被开发用于诊断应用。
方法
本研究制备了重组弓形虫过氧化物还原酶蛋白（rTgPrx），并用于斑点免疫金渗滤法（Dot-IGSS）检测小鼠和孕妇血清中的 IgG 抗体。使用小鼠血清对 rTgPrx-Dot-IGSS 方法进行了建立和优化，并用该方法分析了孕妇血清样本。
结果
分析了 40 份感染弓形虫的小鼠血清样本和 20 份阴性血清样本。rTgPrx-Dot-IGSS 的灵敏度和特异性分别为 97.5%和 100%，与商业 ELISA 试剂盒检测抗弓形虫 IgG 抗体的灵敏度和特异性相当。此外，用商业 ELISA 试剂盒筛选了 540 份孕妇血清样本。用 rTgPrx-Dot-IGSS 分析了 83 份阳性和 60 份阴性血清样本。阳性率为 95.18%，与商业 ELISA 试剂盒相当。
结论
以 rTgPrx 为抗原的 Dot-IGSS 方法可能有助于个体弓形虫感染的诊断。
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