DETECTION OF HUMAN ANTI-ZIKA VIRUS IgG BY ELISA USING AN ANTIGEN FROM in vitro INFECTED VERO CELLS: PRELIMINARY RESULTS.
DETECTION OF HUMAN ANTI-ZIKA VIRUS IgG BY ELISA USING AN ANTIGEN FROM in vitro INFECTED VERO CELLS: PRELIMINARY RESULTS.
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DOI：
10.1590/s1678-9946201658089
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发表时间：
2016-12-08
影响因子：
1.9
通讯作者：
Júnior HF
中科院分区：
文献类型：
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作者：
Sumita LM;Rodrigues JP;Ferreira NE;Felix AC;Souza NC;Machado CM;Júnior HF
寨卡病毒（ZKV）感染在巴西是一个巨大的公共卫生问题，因为受感染母亲所生的新生儿小头畸形的发病率增加。检测母体血清中特异性IgG抗体是诊断ZKV感染及评价其与新生儿小头畸形关系的重要手段。然而，由于巴西没有检测抗ZKV的IgM和IgG抗体的血清学检测方法，我们试图检测来自患者或疑似母亲的血清样本中的特异性IgG，以检测以前的感染，并检测发生在同一地区的黄病毒感染的特异性。从感染的Vero细胞层或培养基中的游离病毒粒子中获得巴西寨卡病毒原生抗原，然后用于ELISA。我们检测了8例经ZKV rna诊断的感染患者（ZKVR）、7例小头畸形新生儿及其产后母亲（MM）、140例登革热病毒igm阳性（DM）和IgG （DG）阳性患者以及100例黄热病（YF）疫苗接种患者的血清。酶联免疫吸附试验结果显示，ZKVR抗体大部分呈阳性（7/8），MM血清ZKV IgG抗体全部呈阳性（7/7）。相反，在接种登革热或黄热病疫苗的患者中观察到交叉反应，包括DM（48/95）、DG（10/45）或YF（3/100）血清样本；然而，这些交叉反应表现出低抗原亲和性，因此6m尿素在很大程度上消除了这种交叉反应性，只剩下少数交叉反应样品（8/140）。基于提取的病毒粒子的ELISA更具特异性，所有ZKVR（8/8）和MM血清ZKV IgG（7/7）阳性，仅发现DM (6/95)， DG（3/45）或YF（4/100）接种血清样本具有临界交叉反应性。该技术（ELISA）可识别zkv感染患者特异性IgG，可能有助于母体RNA病毒清除后先天性感染的诊断或流行病学研究。
Zika virus (ZKV) infection is a huge public health problem in Brazil because of the increased incidence of microcephaly in neonates from infected mothers. Detection of specific IgG antibodies in maternal serum samples constitutes an important approach for diagnosing ZKV infection and evaluating its relationship with neonatal microcephaly. However, as there is no serological test produced in Brazil to detect IgM and IgG antibodies against ZKV, we sought to examine specific IgG in serum samples from patients or suspected mothers to detect previous infection and to test for specificity with regard to flaviviral infections occurring in the same area. Brazilian Zika virus native antigens were obtained from infected Vero cell layers or free virions in the culture medium and then used in ELISA. We tested sera from eight ZKV RNA-diagnosed infected patients (ZKVR), seven neonates with microcephaly and their mothers after delivery (MM), 140 dengue virus IgM-positive (DM) and IgG (DG)-positive patients, and 100 yellow fever (YF)-vaccinated patients. According to the ELISA, ZKVR samples were mostly positive (7/8), and all the MM serum samples were positive for ZKV IgG (7/7). In contrast, cross-reactions for dengue or yellow fever-vaccinated patients were observed, including DM (48/95), DG (10/45) or YF (3/100) serum samples; however, these cross-reactions exhibited low antigen avidity so that 6 M urea largely removed this cross-reactivity, with only a few cross-reacting samples remaining (8/140). ELISA based on extracted virions was much more specific, with all ZKVR (8/8) and MM sera being positive for ZKV IgG (7/7) and only borderline cross-reactivity found for DM (6/95), DG (3/45) or YF (4/100)-vaccinated serum samples. This technique (ELISA) can identify specific IgG in ZKV-infected patients and may be helpful in diagnosing congenital infetions after maternal RNA virus clearance or in epidemiological studies.