Zhang Aiying, Xiu Bingshui, Zhang Heqiu, Li Ning
Beijing Institute of Hepatology, Beijing YouAn Hospital, Capital Medical University, Beijing, China.
Department of Vaccine Engineering, Institute of Basic Medical Sciences, Beijing, China.
J Int Med Res. 2016 Apr;44(2):287-96. doi: 10.1177/0300060515604981. Epub 2016 Feb 1.
To utilize prokaryotic gene expression and protein microarray to develop and evaluate a sensitive, accurate protein microarray assay for detecting antienterovirus antibodies in serum samples from patients with hand, foot and mouth disease (HFMD). Enterovirus 71 (EV71) and coxsackievirus A16 (CA16), two common causative agents for HFMD, were used for assay development.
Serum was collected from patients with HFMD and healthy controls. EV71 and CA16 VP1 and VP3 genes were expressed in transfected Escherichia coli; the resultant VP1 and 3 proteins were used in a microarray assay for human serum EV71 and CA16 immunoglobulin (Ig) M and IgG. To validate the microarray assay, serum samples were tested for EV71 IgM using enzyme-linked immunosorbent assay (ELISA).
Out of 50 patients with HFMD, EV71 IgM and CA16 IgM was detected in 80% and 44% of serum samples, respectively, using protein microarray, and EV71 IgM was detected in 78% of samples using ELISA. Protein microarray and ELISA showed 100% specificity for EV71-IgM detection.
The protein microarray assay developed in the present study shows potential as a sensitive technique for detecting EV71 IgM in serum samples from patients with HFMD.
利用原核基因表达和蛋白质芯片技术开发并评估一种灵敏、准确的蛋白质芯片检测方法，用于检测手足口病（HFMD）患者血清样本中的抗肠道病毒抗体。肠道病毒71型（EV71）和柯萨奇病毒A16型（CA16）是手足口病两种常见的病原体，用于检测方法的开发。
采集手足口病患者和健康对照者的血清。EV71和CA16的VP1和VP3基因在转染的大肠杆菌中表达；所得的VP1和VP3蛋白用于检测人血清中EV71和CA16免疫球蛋白（Ig）M和IgG的蛋白质芯片检测。为验证该蛋白质芯片检测方法，采用酶联免疫吸附测定（ELISA）检测血清样本中的EV71 IgM。
在50例手足口病患者中，使用蛋白质芯片分别在80%和44%的血清样本中检测到EV71 IgM和CA16 IgM，使用ELISA在78%的样本中检测到EV71 IgM。蛋白质芯片和ELISA对EV71-IgM检测的特异性均为100%。
本研究开发的蛋白质芯片检测方法显示出作为一种灵敏技术检测手足口病患者血清样本中EV71 IgM的潜力。
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