常见食源性致病菌基因芯片鉴定技术  

A gene chip method for identifying foodborne pathogenic microorganisms

作　　者：顾鸣[1] 韩伟[1] 关嵘[1] 曹际娟[2] 

机构地区：[1]上海出入境检验检疫局,上海200135 [2]辽宁出入境检验检疫局,沈阳116001

出　　处：《中国卫生检验杂志》2005年第11期1309-1312,共4页Chinese Journal of Health Laboratory Technology

摘　　要：目的:建立鉴定食源性致病菌的基因芯片技术。方法:采用复合PCR方法扩增致病菌特异性DNA片段,结合基因芯片杂交技术鉴定不同的食源性致病菌。结果:本研究完成了采用3对引物的复合PCR扩增;不同致病菌特异性核苷酸片段同时布阵在一张芯片上;标记物质采用了非放射性的生物素,经不同标准菌种、实际检验样品和水平测试样品的考核验证,该鉴定系统灵敏度达620 cfu/g,特异性高,基因芯片质量稳定,该鉴定系统基本含盖了目前食源性致病菌鉴定的需要。结论:常见食源性致病菌基因芯片鉴定技术,可为常规细菌检验方法的最终鉴定提供进一步佐证,尤其在一些培养条件苛刻的致病菌(产单核李斯特菌、弯曲菌等)的鉴定,以及在VITEK仪无法鉴定和手工鉴定判断误差大的情况下,基因芯片方法将发挥其独特的技术性优势,大大提高检验鉴定结果的准确性。Objective：To establish a gene chip method for identifying foodborne pathogenic microorganisms. Methods：Different foodborne pathogenic microorganisms were distinguished by amplification of pathogenic microorganisms specific DNA sequence combined with gene chip hybridization technique. Results：Complex PCR was used to amplify specific genes for different pathogenic microorganisms with three pairs of primers. Specific genes of different foodborne pathogenic microorganism were applied on the same chip and the probe represents different pathogenic microorganisms such as Listeria monocytogenes, Campylobacterjejuni was labeled with DIG. The sensitivity of the method had been repeatedly identified by standard strains. Both conventional and rapid methods were used within practical inspection samples and level test assessment and validation. This rapid method met several reZ quirements concerning accuracy, validation, speed, automation, stabilization, etc. Conclusion： The sensitivity of this identification system can reach 620 cfu/g, and it can be used in the distinguishing for many kinds of hard culture microorganisms such as Listeria monocytogenes, Campylobacterjejuni. This system is a supplement evidence for results obtained from conventional detection methods, and can largely improve the accuracy of inspection on the background of unidentifiable samples beyond of VITEK recognizing range and can minish the estimation error of handwork identification.

关 键 词：食源性致病菌鉴定 基因芯片技术 

分 类 号：R155.3[医药卫生—营养与食品卫生学]