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首页> 外文期刊>Analytical Biochemistry: An International Journal of Analytical and Preparative Methods >Preparation and identification of chicken egg yolk immunoglobulins against human enterovirus 71 for diagnosis of hand-foot-and-mouth disease
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Preparation and identification of chicken egg yolk immunoglobulins against human enterovirus 71 for diagnosis of hand-foot-and-mouth disease

机译：鸡蛋蛋黄免疫球蛋白对人肠道病毒71的制备及鉴定，用于诊断手足口病

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摘要

Human enterovirus 71 (EV71) is one of the major pathogens that causes hand-foot-and mouth disease, and there is an urgent need for rapid diagnosis of EV71 virus infection for early antiviral treatment. The aim of this study was to prepare chicken egg yolk immunoglobulin Y (IgY) for the diagnosis of enterovirus type 71 infection. The antibodies were raised by intramuscular immunization of laying hens with inactivated human EV71 and isolated from the egg yolk by multiple steps of polyethylene glycol 6000 extraction. The average concentration of IgY antibody was 26.60 mg/mL during the whole immunization. After the first immunization, the IgY titer gradually increased, and reached the peak on the 55th days. Meanwhile, the use of western blotting test demonstrated that specific IgY binds specifically to capsid proteins VP2 and VP3 of EV71 virus. Furthermore, a facile one-step method based on turn-on fluorescence sensing was developed by using IgY antibodies, which can detect EV71 virus at low concentrations of 10(4)PFU/mL and was 94.44% coincidence with RT-PCR in 30 clinical samples. These findings indicate that EV71-IgY antibodies are an easily prepared and rich source of antibodies that offers a potential alternative strategy for routine screening of EV71 infection.
机译：人类肠道病毒71（EV71）是导致手足口病的主要病原体之一，迫切需要快速诊断EV71病毒感染对早期抗病毒治疗。本研究的目的是制备鸡蛋蛋黄免疫球蛋白Y（IgY），用于肠病患者71型感染的诊断。通过用灭活的人EV71静置母鸡的肌肉免疫，通过多乙二醇6000萃取从蛋黄分离，从蛋黄蛋白质中分离出抗体。在整个免疫过程中，IgY抗体的平均浓度为26.60mg / ml。在第一次免疫后，IgY滴度逐渐增加，并在第55天达到峰值。同时，蛋白质印迹试验的使用表明，特定IgY特异性地与EV71病毒的衣壳蛋白VP2和VP3结合。此外，通过使用IgY抗体开发了基于导通荧光感测的容易一步法，其可以在10（4）PFU / mL的低浓度下检测EV71病毒，并在30次临床中与RT-PCR重合94.44％样品。这些发现表明EV71-IgY抗体是易于制备的抗体且丰富的抗体来源，其提供常规筛选EV71感染的潜在替代策略。

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