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基于鸡卵黄抗体的粪便抗原捕获ELISA法诊断人体华支睾吸虫病

Chicken IgY-based coproantigen capture ELISA for diagnosis of human opisthorchiasis.

作者信息

Teimoori Salma, Arimatsu Yuji, Laha Thewarach, Kaewkes Sasithorn, Sereerak Piya, Sripa Manop, Tangkawattana Sirikachorn, Brindley Paul J, Sripa Banchob

机构信息

Centre of Research Excellence for Therapeutic Proteins and Antibody Engineering, Department of Parasitology, Faculty of Medicine, Siriraj Hospital, Bangkok 10700, Thailand; WHO Collaborating Centre for Research and Control of Opisthorchiasis (Southeast Asian Liver Fluke Disease), Tropical Disease Research Laboratory, Department of Pathology, Faculty of Medicine, Khon Kaen University, Khon Kaen 40002, Thailand.

WHO Collaborating Centre for Research and Control of Opisthorchiasis (Southeast Asian Liver Fluke Disease), Tropical Disease Research Laboratory, Department of Pathology, Faculty of Medicine, Khon Kaen University, Khon Kaen 40002, Thailand.

出版信息

Parasitol Int. 2017 Aug;66(4):443-447. doi: 10.1016/j.parint.2015.10.011. Epub 2016 Apr 29.

Abstract

Diagnosis of Opisthorchis viverrini infection by conventional stool examination is increasingly difficult due to the low intensity of the infection after several rounds of control programmes in endemic regions as well as coinfections with intestinal flukes. Therefore sensitive and specific diagnostic test is needed. In this study, a coproantigen sandwich ELISA using recombinant O. viverrini cathepsin F (rOv-CF) was developed. This sandwich ELISA employing chicken IgY raised against rOv-CF in combination with rabbit IgG antibody to the somatic O. viverrini antigens showed a lower detection limit (LLD) of 70ng native O. viverrini somatic antigens by spiking the parasite antigens into control feces. When applied to the diagnosis, the IgY-based sandwich ELISA exhibited sensitivity and specificity of 93.3% and 76.7%, respectively, in an investigation of 90 human cases positive or negative for opisthorchiasis. The positive predictive value (PPV) and negative predictive value (NPV) for this coproantigen detection were 66.7% and 95.2%, respectively. This IgY-based sandwich ELISA using parasite cathepsin F detection shows a promising immunodiagnostic alternative for human opisthorchiasis in endemic regions.

摘要

由于流行地区经过几轮防控计划后感染强度较低以及与肠道吸虫的混合感染,通过传统粪便检查诊断华支睾吸虫感染变得越来越困难。因此,需要灵敏且特异的诊断检测方法。在本研究中,开发了一种使用重组华支睾吸虫组织蛋白酶F(rOv-CF)的粪便抗原夹心ELISA。这种夹心ELISA采用针对rOv-CF产生的鸡IgY,并结合针对华支睾吸虫体抗原的兔IgG抗体,通过将寄生虫抗原加入对照粪便中,显示出对70ng天然华支睾吸虫体抗原的较低检测限(LLD)。在对90例华支睾吸虫病阳性或阴性的人类病例进行调查时,基于IgY的夹心ELISA的敏感性和特异性分别为93.3%和76.7%。这种粪便抗原检测的阳性预测值(PPV)和阴性预测值(NPV)分别为66.7%和95.2%。这种基于IgY使用寄生虫组织蛋白酶F检测的夹心ELISA为流行地区人类华支睾吸虫病提供了一种有前景的免疫诊断替代方法。

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