牛和人天然感染巨型片形吸虫排泄/分泌抗原的血清学免疫检测

Rehab Khalil, A. Ibrahim
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引用次数: 1

摘要

传统的片吸虫病诊断方法是基于粪便中卵的鉴定,但这种方法有许多限制,不是一种可靠的方法。采用夹心Elisa法和点印迹法分别对天然感染的牛血清和自然感染的人血清进行巨片吸虫排泄/分泌抗原的免疫检测,并与传统方法进行比较。本研究采集新鲜的巨片吸虫成虫,提取粗排泄/分泌(E/S)抗原。用E/S免疫家兔和小鼠,制备多克隆抗体。纯化兔和小鼠抗片形吸虫抗体IgG部分。抗片形吸虫IgG抗体蛋白含量分别为8.8和5.2mg/ml。采用夹心ELISA法检测248头牛血清中的片形吸虫抗原。同时,采用Dot blot法检测38份人血清中的片形吸虫抗原。结果经粪便寄生虫学检查,26份人标本呈阳性,12份人血清健康。仅在所有感染样本中出现红棕色。这种技术既便宜又节省时间;多个样品可以同时制作,不需要昂贵的实验室设备。综上所示,夹心ELISA和Dot blot检测是较传统方法更可靠的板形吸虫病早期血清诊断工具。意义的声明
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Serological Immunodetection of Fasciola gigantica Excretory/ Secretory Antigens in Naturally Infected Cattle and Human
The traditional diagnostic method of fascioliasis was based on the identification of the eggs in the stool, but this is not a reliable way as it has many restrictions. The present work was evaluated to immunodetect the Excretory/ Secretory antigens of Fasciola gigantica by sandwich Elisa in naturally infected cattle's sera and by dot blot in naturally infected human sera and compare these methods and the traditional methods. In this study, fresh adult Fasciola gigantica worms were collected to extract crude excretory/secretory (E/S) antigens. E/S was used to immunize rabbits and mice to raise polyclonal antibodies. The IgG fraction of rabbit and mouse anti-Fasciola antibodies was purified. The protein content of anti-Fasciola IgG antibody, was 8.8 and 5.2mg/ml, respectively. Sandwich ELISA was performed to detect Fasciola antigens in serum samples collected from 248 cattle. Also, a Dot blot was performed to detect Fasciola antigens in 38 human sera. Results showed that after parasitological stool examination, 26 human samples were positive and 12 human sera were healthy. The red-brown color appeared with all infected samples only. This technique is cheap, and saves time; multiple samples can make at the same time and not require expensive laboratory equipment. In conclusion, the sandwich ELISA and Dot blot assays were more reliable tools for early serodiagnosis of fasciolosis than traditional methods. Significance Statement
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