华南地区常见的8种食源性致病菌检测基因芯片研制  

Development of a DNA microarray for detecting 8 common species of food-borne bacterial pathogens in south China

作　　者：王洪敏[1,2] 何冬梅[1] 周辉[2] 柯碧霞[1] 邓小玲[1] 朱海明[1] 陈经雕[1] 黎薇[1] 杨杏芬[1] 柯昌文[1] 

机构地区：[1]广东省疾病预防控制中心,广东广州510300 [2]解放军第458医院,广东广州510600

出　　处：《南方医科大学学报》2010年第11期2472-2476,共5页Journal of Southern Medical University

基　　金：广东省科技计划项目(2006A20301006)

摘　　要：目的研制能同时检测华南地区常见8种食源性致病菌的oligo基因芯片。针对8种食源性致病菌基因组上的特征性靶区域,设计70mer的寡核苷酸探针。方法分别以检测8种食源性致病菌中每一种菌的检测探针组构建一个亚阵列;然后再将上述的8个亚阵列构建为一个大阵列,并命名为Biosafood-8芯片。用Biosafood-8芯片检测18个种属样品后进行PPR的排序与比较。结果 LM亚阵列检测李斯特菌属的3个李斯特菌LM、Lin和Liv的PPR分别为68.8%、51.8%和59.6%,而其它非李斯特菌属样品的PPR都在43%以下;VC亚阵列检测VC样品的PPR为54.1%,而其它非VC样品的PPR都在17.2%以下;VP亚阵列检测VP样品为66.7%,而其它非VP样品都在24.2%以下;Sal亚阵列检测Sal样品为55.9%,而其它非Sal样品都在50.5%以下;Shi亚阵列检测Shi样品为53.8%,而其它非Shi样品都在36.6%以下;SA亚阵列检测SA样品为65.2%,而其它非SA样品都在22.7%以下;CJ亚阵列检测CJ和CC分别为88.2%和58.8%,而其它非弯曲菌样品都在35.3%以下;EC亚阵列检测EC样品为47.9%,而其它非EC样品都在41.6%以下。结论用Biosafood-8芯片检测18个不同种属来源的已知参考生物样品,从全阵列和芯片上每一个亚阵列两个不同的水平,都证实该芯片能够通过集成的8个目标菌检测探针亚阵列,在整体芯片阵列水平,清晰而又明确地区分目标致病菌样品和非目标致病菌样品,特异而又准确地显示被检测样品的目标菌种属。Objective To prepare a DNA Microarray that can detect 8 commonspecies of food borne bacterial pathogens in south China.Methods 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.Results 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.Conclusion 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.

关 键 词：食源性致病菌 DNA微阵列 寡核苷酸探针 

分 类 号：R155.5[医药卫生—营养与食品卫生学]