金黄色葡萄球菌毒力基因检测及Spa分型研究

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目的研究深圳市南山区2009 2015年医院内患者临床检测标本、医院台面与医护人员双手涂抹样以及食物中毒样本中金黄色葡萄球菌(Staphylococcus aureus,SA)蛋白A基因多态性分型(single locus DNA-sequencing of the repeat region of the Staphylococcus protein A gene,Spa)的特征及毒力基因分布情况。方法采用多重PCR方法检测SA的mec A与fem A基因,PCR方法检测24个毒力基因以及SA蛋白A基因,PCR产物测序后序列上传数据库进行比对。结果 87株SA共检出48株耐甲氧西林金黄色葡萄球菌(methicillin-resistant S.aureus,MRSA),检出率为55.2%。共检出21种毒力基因,检出率>60%的毒力基因有LUKDE(65.5%)、SHE(67.8%)、mpHLG2(85.1%)、HLD(93.1%)和HLA(100.0%)。94.3%菌株携带5个以上毒力基因。三类样本中有11种毒力基因检出率差异有统计学意义,分别是SEB、mpSEC、mpSEJ、mpSEO、TST、HLB、HLD、mpHLG1、mpHLG2,mpETB和PVL。其中mpETB与PVL仅在患者中检出,肠毒素基因在食物中毒株中携带率较高。MRSA与甲氧西林敏感金黄色葡萄球菌(methicillin-sensitive S.aureus,MSSA)的3种毒力基因检出率差异有统计学意义,分别是mpETA、PVL及mpHLG2;MRSA的毒力基因mpETA与PVL检出率明显高于MSSA(P=0.033,P=0.030),而mpHLG2检出率明显低于MSSA(P=0.021)。87株SA分为24种Spa型,患者优势型别为t030(30.0%)与t437(36.7%),医院其他样本优势型别为t091(50.0%),食物中毒优势型别为t127(36.8%)与t091(21.1%)。结论三类样本部分毒力基因检出差异有统计学意义。MRSA与MSSA携带毒力基因差异无统计学意义。三类样本中均存在优势的Spa型别。 Objective To investigate the relationship between the clinical samples of hospital in Nanshan District, Shenzhen, 2015 and the double smears of hospital tabletops and medical personnel and the single locus of Staphylococcus aureus (SA) protein A gene in food poisoning samples. DNA-sequencing of the repeat region of the Staphylococcus protein A gene, Spa) and the distribution of virulence genes. Methods Multiplex PCR was used to detect the mec A and fem A genes in SA. 24 virulence genes and SA protein A gene were detected by PCR. The sequence of the PCR products was sequenced and compared. Results 48 strains of methicillin-resistant S. aureus (methicillin-resistant S. aureus, MRSA) were detected in 87 isolates. The detection rate was 55.2%. A total of 21 virulence genes were detected. LUKDE (65.5%), SHE (67.8%), mpHLG2 (85.1%), HLD (93.1%) and HLA (100.0%) were the virulence genes with the detection rate of> 60%. 94.3% strains carry more than 5 virulence genes. The detection rate of 11 virulence genes in three types of samples was statistically significant, SEB, mpSEC, mpSEJ, mpSEO, TST, HLB, HLD, mpHLG1, mpHLG2, mpETB and PVL respectively. Which mpETB and PVL detected only in patients, enterotoxin gene carrier in food poisoning higher. The detection rates of the three virulence genes of MRSA and methicillin-sensitive S. aureus (MSSA) were statistically different, which were mpETA, PVL and mpHLG2, respectively. The virulence genes mpETA and The detection rate of PVL was significantly higher than that of MSSA (P = 0.033, P = 0.030), but the detection rate of mpHLG2 was significantly lower than that of MSSA (P = 0.021). 87 strains of SA were divided into 24 spa types. The predominant types of patients were t030 (30.0%) and t437 (36.7%), the dominant type of other samples was t091 (50.0%) and the predominant food poisoning type was t127 (36.8% ) And t091 (21.1%). Conclusion The differences of virulence genes in three types of samples were statistically significant. There was no significant difference in the virulence genes between MRSA and MSSA. There are three types of spa in the advantages of the type.
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