肺炎克雷伯菌耐药基因的检测及流行病学研究

Detection of drug-resistant Klebsiella pneumoniae genes and epidemiological research

目的研究医院临床分离的肺炎克雷伯菌的耐药基因以及流行病学特征,提高临床对其认识及诊治水平。方法收集2012年3月-2013年5月临床分离的45株肺炎克雷伯菌,采用纸片琼脂扩散法(K-B)测定其对18种药敏纸片的耐药性,并对β-内酰胺酶、氨基糖苷类药物修饰酶等耐药基因进行PCR扩增,并用DNA测序证实。结果 45株肺炎克雷伯菌对亚胺培南/西司他丁100.0%敏感,对氨苄西林/舒巴坦、头孢西丁、氨曲南、妥布霉素、哌拉西林/他唑巴坦的耐药率均<60.0%,对其余抗菌药物的耐药率均>60.0%;45株肺炎克雷伯菌检出β-内酰胺酶基因最多的为TEM32株检出率为71.1%,其次为SHV10株检出率为22.2%;氨基糖苷类修饰酶阳性检出最多的是aac(3)-Ⅱ39株,检出率为86.7%,其次为aac(6)-Ⅰb 18株、ant(3)-Ⅰ6株,检出率分别为40.0%、13.3%;检出氨基糖苷类修饰酶基因40株,检出率为88.9%,qacE-1-sull 38株,检出率为84.4%。结论耐药基因的肺炎克雷伯菌对临床患者治疗造成了极大困难,因此,有必要加强对产ESBLs肺炎克雷伯菌的分子流行病学监测,从而更好地指导临床,控制肺炎克雷伯菌感染。

OBJECTIVE To study the drug resistant gene of the hospital clinical isolates of K lebsiella pneumoniae （KPN） and its epidemiologic features so as to improve clinical awareness and diagnosis and treatment .METHODS The disk diffusion （K‐B method） method was used for the determination of drug resistance of 45 clinical isolates of KPN isolated during Mar .2012 to May 2013 ,and the drug‐resistant genes such as β‐lactamases ,aminoglycoside modification enzymes were treated by PCR amplification and confirmed by DNA sequencing .RESULTS The sus‐ceptibility test showed the 45 strains of KPN had the 100 .0% sensitivity to imipenem/cilastatin ,the resistance rate of ＆lt;60 .0% to ampicillin/sulbactam ,cefoxitin ,aztreonam ,tobramycin ,piperacillin/tazobactam ,and the re‐sistance rate of ＆gt;60 .0% to other antimicrobials .Among the 45 strains of KPN ,theβ‐lactamase gene was most detected in TEM （32 strains ,the detection rate 71 .1% ） ,followed by SHV （10 ,22 .2% ） .Aminoglycoside modi‐fication enzymes were most detected positive in aac（3）‐Ⅱ （39 ,86 .7% ） ,followed by aac（6）‐Ⅰ b （18 ,40 .0% ） and ant（3）‐Ⅰ （6 ,13 .3% ） .And 40 strains were detected with aminoglycoside modification enzyme gene ,the de‐tection rate was 88 .9% ;38 strains with qacE‐1‐sull were detected ,the detection rate of 84 .4% .CONCLUSION KPN with drug‐resistant genes causes great difficulties in clinical treatment of patients ,therefore ,there is a need to strengthen the monitoring of the molecular epidemiology of ESBLs‐producing KPN ,in order to better guide clinical and control of KPN infections .