Validation of Multiplex Serology for human hepatitis viruses B and C, human T-lymphotropic virus 1 and Toxoplasma gondii
Validation of Multiplex Serology for human hepatitis viruses B and C, human T-lymphotropic virus 1 and Toxoplasma gondii
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DOI：
10.1371/journal.pone.0210407
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发表时间：
2019-01-07
期刊：
影响因子：
3.7
通讯作者：
Waterboer, Tim
中科院分区：
文献类型：
--
作者：
Brenner, Nicole;Mentzer, Alexander J.;Waterboer, Tim
多重血清学是一种高通量技术，用于在一个反应容器中同时检测针对多种病原体的特异性血清抗体。建立了乙肝病毒(乙肝病毒)和丙型肝炎病毒(丙型肝炎病毒)、人类T淋巴细胞病毒1型(HTLV-1)和原虫寄生虫弓形虫(弓形虫)的血清学检测方法，并与已建立的参考检测方法进行了验证。对于每个病原体，有3到5个特定的抗原在大肠杆菌中以GST-Tag融合蛋白的形式重组表达，并在单链血清学中进行检测，即仅限于来自某一特定病原体的抗原的检测。对于四种病原体特异性单链分析中的每一种，使用两种病原体特异性抗原来定义总体血清阳性。在乙肝病毒单链血清学的情况下，基于两个独立的参考板验证了对过去自然感染的检测，两个参考板的灵敏度分别为92.3%和93.0%，特异度均为100%。丙型肝炎病毒和HTLV-1单链血清学检测的敏感性分别为98.0%和95.0%，特异性分别为96.2%和100.0%。建立的弓形虫单链血清学检测方法的敏感性为91.2%，特异性为92.0%。当发展的单链血清学分析方法被纳入一个多重小组(即多重血清学)时，它们基本上保持了它们的特征，其中包含了来自广泛其他病原体的额外抗原。因此，可以将乙肝病毒、丙型肝炎病毒、HTLV-1和弓形虫单链血清学分析有效地合并到为应用于血清流行病学研究而量身定做的多重血清分析板中。
Multiplex Serology is a high-throughput technology developed to simultaneously measure specific serum antibodies against multiple pathogens in one reaction vessel. Serological assays for hepatitis B (HBV) and C (HCV) viruses, human T-lymphotropic virus 1 (HTLV-1) and the protozoan parasite Toxoplasma gondii (T. gondii) were developed and validated against established reference assays. For each pathogen, between 3 and 5 specific antigens were recombinantly expressed as GST-tag fusion proteins in Escherichia coliand tested in Monoplex Serology, i.e. assays restricted to the antigens from one particular pathogen. For each of the four pathogen-specific Monoplex assays, overall seropositivity was defined using two pathogen-specific antigens. In the case of HBV Monoplex Serology, the detection of past natural HBV infection was validated based on two independent reference panels resulting in sensitivities of 92.3% and 93.0%, and specificities of 100% in both panels. Validation of HCV and HTLV-1 Monoplex Serology resulted in sensitivities of 98.0% and 95.0%, and specificities of 96.2% and 100.0%, respectively. The Monoplex Serology assay for T. gondii was validated with a sensitivity of 91.2% and specificity of 92.0%. The developed Monoplex Serology assays largely retained their characteristics when they were included in a multiplex panel (i.e. Multiplex Serology), containing additional antigens from a broad range of other pathogens. Thus HBV, HCV, HTLV-1 and T. gondii Monoplex Serology assays can efficiently be incorporated into Multiplex Serology panels tailored for application in seroepidemiological studies.