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用于检测南方根结线虫的酶联免疫吸附测定(ELISA)的开发。

Development of enzyme linked immunosorbent assay (ELISA) for the detection of root-knot nematode Meloidogyne incognita.

作者信息

Kapur-Ghai J, Kaur M, Goel P

机构信息

Department of Zoology, Punjab Agricultural University, Ludhiana, 141004 India.

Amity Institute of Biotechnology, Amity University, Noida, India.

出版信息

J Parasit Dis. 2014 Sep;38(3):302-6. doi: 10.1007/s12639-013-0246-0. Epub 2013 Jan 26.

Abstract

Root-knot nematodes (Meloidogyne incognita) are obligate, sedentary plant endoparasites that are extremely polyphagous in nature and cause severe economic losses in agriculture. Hence, it is essential to control the parasite at an early stage. For any control strategy to be effective, an early and accurate diagnosis is of paramount importance. Immunoassays have the inherent advantages of sensitivity and specificity; have the potential to identify and quantify these plant-parasitic nematodes. Hence, in the present studies, enzyme-linked immunosorbent assay (ELISA) has been developed for the detection of M.incognita antigens. First an indirect ELISA was developed for detection and titration of anti-M.incognita antibodies. Results indicated as high as 320 K titre of the antisera. Finally competitive inhibition ELISA was developed employing these anti-M.incognita antibodies for detection of M.incognita antigens. Sensitivity of ELISA was 10 fg. Competitive inhibition ELISA developed in the present studies has the potential of being used as an easy, rapid, specific and sensitive diagnostic tool for the detection of M.incognita infection.

摘要

根结线虫(南方根结线虫)是专性定居型植物内寄生线虫,具有极强的多食性,给农业造成严重经济损失。因此,在早期控制这种寄生虫至关重要。对于任何有效的防治策略而言,早期准确诊断至关重要。免疫测定法具有灵敏度和特异性的固有优势,有潜力识别和定量这些植物寄生线虫。因此,在本研究中,已开发出酶联免疫吸附测定法(ELISA)用于检测南方根结线虫抗原。首先开发了一种间接ELISA用于检测和滴定抗南方根结线虫抗体。结果表明抗血清效价高达320K。最后,利用这些抗南方根结线虫抗体开发了竞争抑制ELISA用于检测南方根结线虫抗原。ELISA的灵敏度为10fg。本研究中开发的竞争抑制ELISA有潜力作为一种简便、快速、特异且灵敏的诊断工具用于检测南方根结线虫感染。

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