食品企业污水中细菌的基因芯片快速检测技术

A Genechip Technique for Rapidly Detecting Bacteria in Wastewater of Food Industry

食品企业污水中含有各种无机污物和有机污物,其中夹带的致病菌将导致多种疾病的爆发和流行,严重威胁着人类的健康。传统的细菌分离、培养和鉴定技术操作繁琐且耗时长(一般需4～7d)。为了快速、准确地检测食品企业污水中存在的细菌,建立了一种采用基因芯片技术对食品企业污水中常见细菌检测和鉴定的实验方法(需时4h)。实验中设计了8对特异引物并成功分成2组混合引物进行多重 PCR 反应:引物Ⅰ为 Kp+HyA+InvA+ipaH,引物Ⅱ为 Cadc+Oprl+Ent+23SrRNA,效果良好。实验确定了适宜的PCR 反应循环数为35个循环,适宜的杂交温度为48℃。用实验制备的基因芯片对模拟水样检测结果的准确率达100%,说明该基因芯片对目的细菌特征基因的检测结果是可靠的。用该方法对食品企业污水水样进行检测具有高准确率,为今后更大规模的检测研究奠定了基础。

There are all kinds of organic and inorganic contaminants in the wastewater of food industry. The pathogens contained in it can result in epidemic diseases to threaten the human health seriously. The traditional operation of bacterium separation, incubation and identification is convoluted and time-consuming （usually taking about 4 to 7 days）. To detect the bacteria in wastewater of food industry quickly and accurately, a rapid produce based on the genechip technique has been set up （accomplishing within 4 hours）. In the experiment, eight pairs of special primers were designed and successfully divided into two sets, Kp＋HlyA＋lnvA＋ipaH and Cadc＋Oprl＋Ent＋23SrRNA, to perform the multiple polymerase chain reaction （PCR） effectively. Through the experiment, the suitable PCR rotation numbers was 35 cycles and the suitable hybridisation temperature was 48℃. In the simulation detection, the accuracy of the resultsby using the genechip technique reached 100%. It meant that this technique was reliable and able to be used in accurately detecting wastewater samples of food industry. This technique is likely promising for more extensive application in the future.