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应用刚地弓形虫排泄/分泌抗原对脑弓形虫病和 HIV 感染患者的脑脊液进行免疫诊断。

Immunodiagnosis in cerebrospinal fluid of cerebral toxoplasmosis and HIV-infected patients using Toxoplasma gondii excreted/secreted antigens.

机构信息

Laboratório de Parasitologia, Instituto Adolfo Lutz, CEP 01246-902 São Paulo, SP, Brazil.

出版信息

Diagn Microbiol Infect Dis. 2011 Nov;71(3):279-85. doi: 10.1016/j.diagmicrobio.2011.07.008. Epub 2011 Sep 9.

Abstract

Cerebral toxoplasmosis is the most common neurologic opportunistic infection in HIV-infected patients. Excretory-secretory antigens (ESA) are the majority of the circulating antigens in sera from hosts with acute toxoplasmosis, and their usefulness as antigens has been shown. This study considered whether it could find anti-ESA antibodies in cerebrospinal fluid (CSF) and whether these antibodies can be markers of active infection. Samples of CSF from 270 HIV-infected patients were analyzed and divided into 3 groups according to the presence or absence of active toxoplasmosis. Group I: 99 patients with cerebral toxoplasmosis; group II: 112 patients with other opportunistic neurologic diseases and seropositive for toxoplasmosis; and group III: 59 patients with other opportunistic neurologic diseases and seronegative for toxoplasmosis. Toxoplasma gondii ESA and a crude tachyzoite antigen were used as antigens using ELISA and immunoblotting. The statistical analysis was done using the F test and unpaired Student's t test. Crude tachyzoite antigen: mean ELISA-relative values ± standard error for CSF of groups I and II were 7.0 ± 0.27 and 3.9 ± 0.19, respectively. Variance analysis revealed that results of both groups of patients were statistically different (1.80, P = 0.0025). The difference between the mean results was 3.0 ± 0.3, and the Student's t test value was 9.41 (P = 0.0001). Samples from groups I and II were reactive by immunoblotting, with similar intensities. In ESA-ELISA, the mean for group I was 9.0 ± 0.39. Group II showed a mean value of 2.7 ± 0.12. Both groups were statistically different (9.16, P < 0.001). However, in ESA, the difference between the mean results was higher (6.2 ± 0.39) and the Student's t test value was 16.04 (P < 0.0001). Similar results were shown in immunoblotting where a CSF sample from group I reacted well with ESA, and the sample from a group II patient failed to do so. The mean ELISA-relative value of the control group (group III) was 0.5 ± 0.09 for the first antigen and 0.4 ± 0.22 for the second. ESA-ELISA and/or immunoblotting of CSF samples can be used for diagnosis of cerebral toxoplasmosis in association with clinical, serologic, and radiological information, thus providing a simple straightforward methodology, particularly suitable in countries with high prevalence of latent toxoplasmosis in the general population.

摘要

脑弓形体病是 HIV 感染患者中最常见的神经机会性感染。排泄-分泌抗原(ESA)是急性弓形体病宿主循环抗原中的大多数,其作为抗原的有用性已得到证实。本研究考虑是否能在脑脊液(CSF)中找到抗 ESA 抗体,以及这些抗体是否可以作为活动性感染的标志物。分析了 270 例 HIV 感染患者的 CSF 样本,并根据是否存在活动性弓形体病将其分为 3 组。第 I 组:99 例脑弓形体病患者;第 II 组:112 例患有其他机会性神经疾病且弓形体病血清阳性患者;第 III 组:59 例患有其他机会性神经疾病且弓形体病血清阴性患者。使用 ELISA 和免疫印迹法,用刚地弓形体 ESA 和粗线状体抗原作为抗原。使用 F 检验和未配对学生 t 检验进行统计分析。粗线状体抗原:第 I 组和第 II 组 CSF 的 ELISA-相对值平均值±标准误差分别为 7.0±0.27 和 3.9±0.19。方差分析表明两组患者的结果均具有统计学差异(1.80,P=0.0025)。两组平均结果的差异为 3.0±0.3,学生 t 检验值为 9.41(P=0.0001)。第 I 组和第 II 组的样本通过免疫印迹法呈反应性,强度相似。在 ESA-ELISA 中,第 I 组的平均值为 9.0±0.39。第 II 组的平均值为 2.7±0.12。两组均具有统计学差异(9.16,P<0.001)。然而,在 ESA 中,平均结果的差异更高(6.2±0.39),学生 t 检验值为 16.04(P<0.0001)。免疫印迹也显示出相似的结果,第 I 组的 CSF 样本与 ESA 反应良好,而第 II 组患者的样本则没有反应。对照组(第 III 组)的 ELISA-相对平均值分别为第一抗原的 0.5±0.09 和第二抗原的 0.4±0.22。CSF 样本的 ESA-ELISA 和/或免疫印迹可与临床、血清学和影像学信息一起用于脑弓形体病的诊断,从而提供一种简单直接的方法,特别适用于一般人群中潜伏性弓形体病流行率较高的国家。

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