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目的利用毛细管电泳技术和多位点可变数目串联重复序列分析方法(MLVA)对引起一起食物中毒的金黄色葡萄球菌进行分子分型,了解毒株的肠毒素类型,为金黄色葡萄球菌的流行病学调查和溯源提供参考数据。方法以食物中毒分离出的10株金黄色葡萄球菌的基因组为模板,选用8个可变数目串联重复序列(VNTR)位点进行PCR扩增,扩增产物进行毛细管电泳分析,利用BioCalculator软件对电泳图谱进行分析,产生的数据与MLVA数据库提供信息进行比对。同时测定毒株的肠毒素类型。结果 10株菌的8个VNTR位点的PCR扩增产物一致,VNTR09_01、VNTR61_01、VNTR61_02、VNTR67_01、VNTR21_01、VNTR24_01、VNTR63_01和VNTR81_01扩增出的条带大小分别为373、361、328、279、845、354、394和658 bp。其VNTR重复数目为15、2、2、2、35、8、2和7。10株菌的MLVA型(MT)与目前MLVA数据库公布的3892个MT均不相同,是一种新的MLVA型别,10株不同来源金黄色葡萄球菌均产肠毒素A和肠毒素B。结论 10株金黄色葡萄球菌的肠毒素特性和MLVA分型结果均一致,此次食物中毒分离出的10株菌具有高度同源性。
OBJECTIVE: To characterize the type of enterotoxigenic strains of Staphylococcus aureus causing food poisoning by capillary electrophoresis and multi-locus variable number tandem repeat analysis (MLVA) Provide the reference data for the medical investigation and traceability. Methods The genomes of 10 Staphylococcus aureus isolates from food poisoning were used as templates. The 8 VNTR sites were selected for PCR amplification. The amplified products were analyzed by capillary electrophoresis and analyzed by BioCalculator software. Atlas was analyzed and the data generated was aligned with the information provided by the MLVA database. Simultaneous determination of the strain’s enterotoxin type. Results The PCR products of 8 VNTR loci in 10 isolates were identical. The amplified bands of VNTR09_01, VNTR61_01, VNTR61_02, VNTR67_01, VNTR21_01, VNTR24_01, VNTR63_01 and VNTR81_01 were 373, 361, 328, 279, 845 , 354,394 and 658 bp. The MLVA type (MT) with VNTR repeats of 15, 2, 2, 2, 35, 8, 2 and 7.10 strains differ from the 3892 MTs currently published in the MLVA database and is a new type of MLVA Do not, 10 different sources of Staphylococcus aureus produce enterotoxin A and enterotoxin B. Conclusions The enterotoxin characteristics of 10 strains of Staphylococcus aureus are consistent with the results of MLVA typing. The 10 strains isolated from food poisoning have high homology.