Wang Hong-min, He Dong-mei, Zhou Hui, Ke Bi-xia, Deng Xiao-ling, Zhu Hai-ming, Chen Jing-diao, Li Wei, Yang Xing-fen, Ke Chang-wen
Guangdong Provincial Center for Disease Control and Prevention, Guangzhou 510300, China.
Nan Fang Yi Ke Da Xue Xue Bao. 2010 Nov;30(11):2472-6.
To prepare a DNA Microarray that can detect 8 common species of food borne bacterial pathogens in south China.
All the 70mer oligo probes were designed on the characteristic genome loci of the 8 species of food borne bacterial pathogens. Eight subarrays corresponding to the 8 food borne bacterial pathogens were spotted onto the slide and integrated into a pan-array on the chip. A number of identified and known bacterial samples from the storage bank were selected for the validation test.
Based on the PPR ranking, for LM sub-array, the PPR of the 3 Listeria bacteria LM, Lin and Liv was 68.8%, 51.8% and 59.6%, respectively, while that of the non-Listeria bacterial samples was all below 43%. For VC sub-array, the PPR of VC sample was 54.1% and that of the non-VC bacterial samples was lower than 17.2%. For VP sub-array, the PPR was 66.7% for VP sample and below 24.2% for non-VP bacterial samples. For Sal sub-array, the PPR was 55.9% for Sal sample and below 50.5% for non-Sal bacterial samples. For Shi sub-array, the PPR of Shi sample and the non-Shi bacterial samples was 53.8% and below 36.6%, respectively. For SA sub-array, the PPR of SA sample and non-SA bacterial samples was 65.2% and below 22.7%, respectively. For CJ sub-array, the PPR of the 2 Campylobacter bacteria CJ and CC were 88.2% and 58.8%, respectively, and that of the non-Campylobacter bacterial samples was lower than 35.3%. For EC sub-array, the PPR of EC sample was 47.9%, and that of the non-EC bacterial samples was lower than 41.6%. Evaluation of the Biosafood-8 chip developed in this study by 18 biological samples from different origins demonstrated its good specificity and accuracy in the identification of the pathogens.
The chip we developed can clearly differentiate the target food borne pathogenic bacteria and non-target bacteria and allows specific and accurate identification of the species of the tested bacteria isolates.
制备一种能够检测华南地区8种常见食源性细菌病原体的DNA微阵列。
针对8种食源性细菌病原体的特征基因组位点设计所有70mer寡核苷酸探针。将对应8种食源性细菌病原体的8个子阵列点样到载玻片上，并整合到芯片上的一个泛阵列中。从菌种保藏库中选取一些已鉴定的已知细菌样本进行验证测试。
基于PPR排名，对于李斯特菌（LM）子阵列，3种李斯特菌（LM、Lin和Liv）的PPR分别为68.8%、51.8%和59.6%，而非李斯特菌细菌样本的PPR均低于43%。对于副溶血性弧菌（VC）子阵列，VC样本的PPR为54.1%，非VC细菌样本的PPR低于17.2%。对于小肠结肠炎耶尔森菌（VP）子阵列，VP样本的PPR为66.7%，非VP细菌样本的PPR低于24.2%。对于沙门氏菌（Sal）子阵列，Sal样本的PPR为55.9%，非Sal细菌样本的PPR低于50.5%。对于志贺氏菌（Shi）子阵列，Shi样本和非Shi细菌样本的PPR分别为53.8%和低于36.6%。对于金黄色葡萄球菌（SA）子阵列，SA样本和非SA细菌样本的PPR分别为65.2%和低于22.7%。对于弯曲杆菌（CJ）子阵列，2种弯曲杆菌（CJ和CC）的PPR分别为88.2%和58.8%，而非弯曲杆菌细菌样本的PPR低于35.3%。对于大肠埃希氏菌（EC）子阵列，EC样本的PPR为47.9%，非EC细菌样本的PPR低于41.6%。用来自不同来源的18个生物样本对本研究开发的Biosafood-8芯片进行评估，结果表明其在病原体鉴定方面具有良好的特异性和准确性。
我们开发的芯片能够清晰地区分目标食源性病原体和非目标细菌，并能特异性、准确地鉴定被测细菌分离株的种类。
