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使用针对绵羊肝片吸虫排泄/分泌抗原的多克隆抗体评估不同免疫技术的诊断潜力。

Evaluation of the diagnostic potential of different immunological techniques using polyclonal antibodies against Fasciola gigantica excretory/secretory antigens in sheep.

作者信息

El Amir Azza, Rabee Ibraheem, Kamal Nour, El Deeb Somaya

机构信息

Zoology Department, Faculty of Science, Cairo University, Egypt.

出版信息

Egypt J Immunol. 2008;15(1):65-74.

Abstract

The early detection of Fasciola antigens (Ags) in serum or stool could be more valuable in diagnosis as early treatment would be applied before irreparable damage occurs. In this study, fresh adult Fasciola gigantica (F. gigantica) worms were cultivated for 16 hrs. Excretory/secretory (E/S) Ags were extracted from the culturemedium and used to raise rabbit antibodies (Abs) to Fasciola. The purified Abs were then used in sandwich ELISA (S-ELISA) to detect Fasciola Ags in serum and stool samples from a total of 152 sheep, and sandwich-Dot-ELISA (S-D-ELISA) for the serum samples. Gross inspection of liver for flukes or other parasites was performed and results of parasitological stool examination were recorded. Accordingly, sheep were divided into healthy control group (25 sheep), Fasciola positive group (97 sheep) and other helminthic infection groups (30 sheep). S-ELISA for serum samples showed 91.9% sensitivity and 89% specificity. Fasciola Ags, detected in serum of sheep by S-D-ELISA, showed 97.2% sensitivity and 95% specificity and coproantigens detected by S-ELISA, showed 95.8% sensitivity and 92.7% specificity. Although, the specificity of stool examination was higher than that recorded for serum, the sensitivity of ELISA techniques to diagnose Fasciola Ags was higher than that recorded for parasitological examination. It is concluded that, S-D-ELISA has better sensitivity and specificity than S-ELISA for both stool and serum, and may prove useful for field applications.

摘要

在血清或粪便中早期检测肝片吸虫抗原(Ags)对于诊断可能更有价值,因为在不可修复的损伤发生之前就可以进行早期治疗。在本研究中,将新鲜的成年巨片吸虫(F. gigantica)虫体培养16小时。从培养基中提取排泄/分泌(E/S)抗原,并用于制备兔抗肝片吸虫抗体(Abs)。然后将纯化的抗体用于夹心ELISA(S-ELISA)以检测总共152只绵羊的血清和粪便样本中的肝片吸虫抗原,并用于血清样本的夹心斑点ELISA(S-D-ELISA)。对肝脏进行肉眼检查以查找吸虫或其他寄生虫,并记录粪便寄生虫学检查结果。据此,将绵羊分为健康对照组(25只绵羊)、肝片吸虫阳性组(97只绵羊)和其他蠕虫感染组(30只绵羊)。血清样本的S-ELISA显示敏感性为91.9%,特异性为89%。通过S-D-ELISA在绵羊血清中检测到的肝片吸虫抗原显示敏感性为97.2%,特异性为95%,通过S-ELISA检测到的粪抗原显示敏感性为95.8%,特异性为92.7%。虽然粪便检查的特异性高于血清检查,但ELISA技术诊断肝片吸虫抗原的敏感性高于寄生虫学检查。结论是,S-D-ELISA对于粪便和血清的敏感性和特异性均优于S-ELISA,可能在现场应用中证明是有用的。

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