免疫分离技术在生鲜乳致病菌检测中的研究进展

doi:10.3864/j.issn.0578-1752.2015.S.006

摘要/Abstract

摘要： 生鲜乳作为原料乳，其微生物含量是影响生鲜乳新鲜度和产品货架期的重要因素，对生产优质乳及乳制品具有重要意义。生鲜乳中可能存在的金黄色葡萄球菌、单核增生李斯特菌、沙门氏菌及大肠杆菌等致病菌严重威胁公共健康。建立快速灵敏高特异性的检测方法对于保护公共健康非常重要。目前，国内外主要通过传统培养法、ELISA、LAMP及qPCR等方法开展生鲜乳中主要致病菌检测相关研究。上述检测方法对未经前增菌的样品检测灵敏度较低，极大限制了生鲜乳样品中主要致病微生物的快速检测。免疫磁分离技术可以在短时间内高效富集样品中的目的菌，与现代检测技术相结合，可以缩短检测周期，提高检测灵敏度。该方法已广泛应用于金黄色葡萄球菌、单核增生李斯特菌、沙门氏菌、大肠杆菌、禽分枝杆菌、空肠弯曲杆菌、幽门螺旋杆菌、结肠炎耶尔森杆菌及沙眼衣原体等病原微生物的分离检测。免疫磁珠技术与分光光度计、显色培养基和荧光定量PCR、荧光免疫层析、胶体金试纸条、real time PCR相结合极大的缩短了大肠杆菌的检测和分析时间。该技术与显色培养基、比浊分析法、免疫荧光检测和环等温介导技术相结合可实现金黄色葡萄球菌的快速检测。其与qPCR、酶联免疫法、荧光显色成像及环介导等温扩增技术相结合极大提高了沙门氏菌的检测效率。其与流式细胞仪、选择性培养基及荧光定量PCR相结合可有效的缩短单核增生李斯特菌的分离检测时间。然而该技术在生产及应用上还存在检测成本高、适用性受限等一些问题。文章就免疫分离技术在生鲜乳中主要致病菌检测方面的研究进展进行了综述，并在此基础上分析了该技术在生鲜乳致病菌检测方面存在的问题及发展方向，为该技术的应用发展提供一定的参考。

关键词: 免疫分离技术, 生鲜乳, 致病菌检测

Abstract: Microbial community in raw milk is an important factor influencing quality and safety of the dairy products, as it reflects hygienic conditions during milking and storage of raw milk in the bulk tank. Foodborne pathogens of high public health concern, including Staphylococcus aureus, Listeria monocytogenes, Salmonella, Bacillus cereus and Escherichia coli, seriously affected human health. Traditional culture-based methods, ELISA, LAMP and PCR are used for detection of pathogens. Detection sensitivity of existing methods are low without pre-enrichment, limiting the methods directly used for rapid detection of pathogen in raw milk. Immunomagnetic separation has been suggested to reduced the analysis time and improved the sensitivity detection of pathogenic microorganisms. The methods were successfully used to detect Staphylococcus aureus, Listeria monocytogenes, Bacillus cereus, Escherichia coli, Mycobacterium avium, Campylobacter jejuni, Helicobacter pylori, Campylobacter spp, Yersinia spp and Chlamydia trachomatis. Immunomagnetic separation combined with spectrophotometer, Chrom agar, real-time PCR, fluorescence immunochr omatographic assay or colloidal gold lateral flow assay were effective for detection and isolation of Escherichia coli. Immunomagnetic separation combined with Chrom agar, colorimetric detection, fluorescence immunochr omatographic assay and LAMP were effective for detection and isolation of Staphylococcus aureus. Immunomagnetic separation combined with qPCR, ELISA, quantum dots and LAMP were effective for detection and isolation of Salmonella spp. Immunomagnetic separation combined with flow cytometry, selective medium and real time PCR were effective for detection and isolation of Listeria monocytogenes. Recent progress in the application of immunomagnetic separation technology for the mainly pathogens in raw milk was reviewed, and the problems and development trends about immunomagnetic separation were analyzed and discussed in the paper.

Key words: immunomagnetic separation, raw milk, pathogen detection

兰欣怡，刘慧敏，郑楠，李发弟，王加启. 免疫分离技术在生鲜乳致病菌检测中的研究进展[J]. 中国农业科学, 2015, 48(S): 49-57.

LAN Xin-yi, LIU Hui-min, ZHENG Nan, LI Fa-di, WANG Jia-qi. Immunomagnetic Separation for Foodborne Pathogen Detection in Raw Milk[J]. Scientia Agricultura Sinica, 2015, 48(S): 49-57.

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