骨科创伤患者伤口分泌物金黄色葡萄球菌毒力基因分析

Analysis of virulence genes of Staphylococcus aureus in wound secretion of orthopedic trauma patients

目的了解骨科创伤患者伤口分泌物金黄色葡萄球菌(SAU)分离株的毒力基因分布情况,为防控SAU的医院内感染提供依据。方法收集2018年10月至2019年12月东莞市中医院骨科创伤患者伤口分泌物标本80份,经细菌培养分离出80株SAU菌株,根据感染位置深浅将其分为浅表感染组和深部感染组,每组40株。采用聚合酶链反应(PCR)进行毒力基因检测,观察浅表感染组和深部感染组SAU菌株的毒力基因分布情况,比较2组SAU菌株毒力基因检出率的差异。结果80株SAU共检出20种毒力基因,分别为黏附素基因bbp、clfA、clfB、cna、ebpS、fnbA、sdrC、sdrD、sdrE、icaA、map,溶血毒素基因hla、hlb、hld、hlg,杀白细胞素基因lukE、lukM、pvl,侵袭毒素基因splB、ssp;其中各种毒力基因的检出情况分别为:黏附素基因bbp 78株(97.5%)、clfA 74株(92.5%)、clfB 75株(93.8%)、cna 66株(82.5%)、ebpS 75株(93.8%)、fnbA 80株(100.0%)、sdrC 56株(70.0%)、sdrD 40株(50.0%)、sdrE 77株(96.2%)、icaA 78株(97.5%)、map 72株(90.0%),溶血毒素基因hla 80株(100.0%)、hlb 78株(97.5%)、hld 74株(92.5%)、hlg 72株(90.0%),杀白细胞素基因lukE 71株(88.8%)、lukM 6株(7.5%)、pvl 19株(23.8%),侵袭毒素基因splB 14株(17.5%)、ssp 66株(82.5%)。浅表感染组及深部感染组fnbA、hla基因检出率均为100.0%(40/40);深部感染组SAU的clfA、pvl、ssp基因检出率高于浅表感染组(P<0.05);深部感染组与浅表感染组SAU的bbp、clfB、cna、ebpS、sdrC、sdrD、sdrE、icaA、map、hlb、hld、hlg、lukE、lukM及splB基因检出率比较差异无统计学意义(P>0.05)。结论骨科创伤患者伤口分泌物分离SAU检出的20种毒力基因在浅部感染和深部感染患者均有检出,其中clfA基因、pvl基因和ssp基因可能与感染深度相关。

Objective To investigate the distribution of virulence gene of Staphylococcus aureus in wound secretion of orthopedic trauma patients,and to provide evidence for the prevention and control of nosocomial infection of Staphylococcus aureus.Methods Eighty specimen of wound secretion of orthopedic trauma patients in Department of Osteology,Dongguan TCM Hospital from October 2018 to December 2019 were collected.Eighty strains Staphylococcus aureus were isolated by bacterial culture and they were divided into superficial infection group and deep infection group,with 40 strains in each group.The virulence genes of Staphylococcus aureus were detected by polymerase chain reaction.The detection rate and distribution of virulence genes of Staphylococcus aureus were compared between the two groups.Results A total of 20 virulence genes were detected from 80 strains of Staphylococcus aureus,including adhesin gene(bbp,clfA,clfB,cna,ebpS,fnbA,sdrC,sdrD,sdrE,icaA,map),hemolytic toxin gene(hla,hlb,hld,hlg),leukocidin gene(lukE,lukM,pvl),invasive toxin gene(splB,ssp).Among them,78(97.5%)strains were bbp,74(92.5%)strains were clfA,75(93.8%)strains were clfB,66(82.5%)strains were cna,75(93.8%)strains were ebpS,80(100.0%)strains were fnbA,56(70.0%)strains were sdrC,40(50.0%)strains were sdrD,77(96.2%)strains were sdrE,78(97.5%)strains were icaA,72(90.0%)strains were map of adhesin gene;80(100.0%)strains were hla,78(97.5%)strains were hlb,74(92.5%)strains were hld,72(90.0%)strains were hlg of hemolytic toxin gene;71(88.8%)strains were lukE,6(7.5%)strains were lukM,19(23.8%)strains were pvl of leukocidin gene;14(17.5%)strains were splB,66(82.5%)strains were ssp of invasive toxin gene.The detection rates of fnbA and hla genes in both superficial infection group and deep infection group were 100.0%(40/40).The detection rates of clfA,plv and ssp genes of Staphylococcus aureus in the deep infection group were higher than those in the superficial infection group(P<0.05);there was no significant difference in the detection rates of bbp,clfB,cna,ebpS,sdrC,sdrD,sdrE,icaA,map,hlb,hld,hlg,lukE,lukM and splB genes of Staphylococcus aureus between the deep infection group and the superficial infection group(P>0.05).Conclusion The 20 virulence genes of the Staphylococcus aureus from the wound secretions of orthopedic trauma patients were detected in both superficial infection and deep infection patients,among which clfA,pvl and ssp genes may be correlated with the depth of infection.