铜绿假单胞菌金属酶及整合酶的检测

Detection of Metallo-β-Lactamases and Integrasesin from Pseudomonas Aeruginosa Isolates

目的：了解哈尔滨医科大学附属第二医院铜绿假单胞菌（Pa）株中金属β-内酰胺酶及整合酶携带情况。方法采用改良 Hodge法、双纸片协同法、组合纸片法进行金属酶表型检测，PCR法检测金属酶基因型以及整合酶Ⅰ、整合酶Ⅱ和整合酶Ⅲ。将 IMP金属酶全基因PCR产物进行纯化，测序再进行Blast比对分析。结果62株 Pa经金属β-内酰胺酶改良 Hodge法检出3株、双纸片协同法检出4株，组合纸片法检出4株，PCR法检测出4株（6.45％，4／62）金属酶阳性菌株，经全基因测序结果为 IMP-1型，且全部为亚胺培南耐药菌株10％（4／40）；未检测出 VIM，SPM和 GIM型金属酶；PCR检测整合酶Ⅰ在亚胺培南耐药菌株中阳性率为40％（16／40），在亚胺培南敏感菌株中检出率为22.73％（5／22）；所有菌株均未检出整合酶Ⅱ，Ⅲ。结论哈尔滨医科大学附属第二医院存在 IMP-1型金属酶阳性的Pa，这些菌株多为多重耐药细菌，且携带Ⅰ类整合子，该院应做好防范工作，防止金属酶和整合子的传播。

Objective To investigate the situation of Pseudomonas aeruginosa strains carrying metallo-β-lactamases and integrases in the Second Affiliated Hospital of Harbin Medical University.Methods The phenotype of metallo-β-lactamases were detected by modified Hodge test,double-disc synergy and combination paper method,respectvily.PCR method was used to detecte metallo-β-lactamases genotypes and the integrationⅠ,Ⅱ and Ⅲ.The PCR products of the whole length bla-IMP gene were purified,sequenced and analyzed by Blast.Results Among 62 Pseudomonas aeruginosa strains,metallo-β-lactamases phenotype of 3 by modified hodge test,4 by double-disc synergy test,4 by combination paper method and were all positive 4 of IMP-1 type metallo-β-lactamases of Pseudomonasaeruginosa strains （10%,4/40）were positives by PCR.Inte-grase Ⅰ of 16（40 %,16/40）strains were positives by PCR method in IMP-resistant Pseudomonasaeruginosa,and integraseⅠ of 22.73 %（5/22）were positives in IMP-sensitive Pseudomonasaeruginosa.No other metallo-β-lactamases,integraseⅡandⅢ were detected in this study.Conclusion IMP-type metallo-β-lactamase existed in IMP-resistant Pseudomonasaerugi-nosa isolates of the Second Affiliated Hospital of Harbin Medical University.Most strains carried integraseⅠ,and other resistance mechanisms may be associated with multi-drug resistance,so it is important to prevent the Pseudomonasaeruginosa strains which carried metallo-β-lactamases and integrons widely spread in the hospital.