苹果浓缩汁中耐热菌的间接ELISA快速检测方法

doi:10.3864/j.issn.0578-1752.2011.22.014

摘要/Abstract

摘要： 【目的】研究果汁中耐热菌的免疫化学快速检测方法。【方法】以果汁中分离的耐热菌免疫家兔获得抗体，采用酶联免疫吸附法（enzyme-linked immunosorbent assay, ELISA）原理建立果汁中耐热菌快速检测方法。【结果】首次成功建立并优化了耐热菌间接ELISA快速检测方法，检测限为5×10 4CFU/mL。实际检测时，通过对果汁样品集菌及预增菌，应用本研究建立的耐热菌间接ELISA快速检测方法，可以在24 h内给出果汁中耐热菌是否超过1 CFU/10 mL的检测结果，满足国内外果汁标准中耐热菌不超过1 CFU/10 mL的要求，检测时间比目前国内外普遍采用的KFL（Krueger Food Laboratories）法缩短3-4 d，检测结果与KFL法完全相符。【结论】本研究建立的果汁中耐热菌间接ELISA快速检测方法快速、简便、准确，具有良好的特异性、敏感性和重复性。

关键词: 苹果浓缩汁, 耐热菌, 酶联免疫吸附法, 快速检测

Abstract: 【Objective】 An establishment of a rapid and accurate indirect ELISA for detection of Alicyclobacillus acidoterrestris in apple juice concentrate(AJC) was studied. 【Method】 Polyclonal antibody was generated by immunizing rabbits with A. acidoterrestris separated from AJC and indirect enzyme-linked immunosorbent assay was performed to analyze the antigenicity of A. acidoterrestris and its spores. 【Result】 The new indirect ELISA was used successfully in detecting A. acidoterrestris for the first time, and the detection limit was 5×104 CFU/mL. In the detection of AJC, the samples were filter sterilized and enriched, followed by the indirect ELISA assay. This method could be performed in less than 24 h, and the minimum detection limit has reached the international standard for detecting A. acidoterrestris, which AJC is less than 1 CFU/10 mL. The result gained by indirect ELISA was in accord with that by KFL, a conventional culture assay. 【Conclusion】 The developed indirect ELISA has good specificity, sensitivity and reproducibility for the detection of A. acidoterrestris in juice concentrate, which is accurate, convenient, and quick.

Key words: apple juice concentrate, A. acidoterrestris, enzyme-linked immunosorbent assay, rapid detection

李建科, 王峰, 夏凯. 苹果浓缩汁中耐热菌的间接ELISA快速检测方法[J]. 中国农业科学, 2011, 44(22): 4669-4677.

LI Jian-Ke, WANG Feng, XIA Kai. An Indirect and Accurate ELISA for Detection of Alicyclobacillus acidoterrestris in Apple Juice Concentrate[J]. Scientia Agricultura Sinica, 2011, 44(22): 4669-4677.

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