产KPC肺炎克雷伯菌检测方法构建与耐药机制

发布时间：2018-05-19 21:22

本文选题：碳青酶烯酶 + 肺炎克雷伯菌　；参考：《安徽医科大学学报》2017年08期

【摘要】：目的探讨产肺炎克雷伯型碳青霉烯酶(KPC)肺炎克雷伯菌检测方法以及肺炎克雷伯菌对碳青霉烯类抗生素的耐药机制、可能的治疗药物。方法选取K-B法证实的耐碳青霉烯肺炎克雷伯菌36株纳入本研究,使用改良Hodge试验、Carba NP试验对菌株进行验证,随后采用PCR扩增、产物琼脂糖凝胶电泳实验,并对电泳阳性条带进行基因测序鉴定。结果 K-B法药敏试验显示36株耐碳氢烯类肺炎克雷伯菌,随后行Hodge验证,33株(91.7%)是产KPC肺炎克雷伯菌,对碳氢烯类耐药而对替加环素及米诺环素仍然敏感。对Hodge试验阳性33株再行Carba NP试验确证,其中27株(87.9%)阳性,6株(12.1%)阴性;对Hodge试验阳性33株行PCR扩增及电泳实验,共有27株出现目标条带(340bp)提示阳性,与Carba NP试验结果相一致。对出现目标条带的27株肺炎克雷伯菌再进行KPC-2耐药基因PCR扩增产物测序,均为KPC-2型耐药基因。结论 K-B法药敏试验加随后改良Hodge试验验证对于筛查、发现KPC肺炎克雷伯菌具有较高的符合度,为发现KPC肺炎克雷伯菌有效筛查方法。K-B法药敏试验证实对KPC肺炎克雷伯菌,替加环素等可能是有效治疗的药物之一。Hodge试验检测的KPC肺炎克雷伯菌,存在12.1%(6/33)的假阳性率。Carba NP试验及PCR扩增检测一致性好,具有确证产KPC肺炎克雷伯菌作用。随后的测序研究结果表明检测出的KPC肺炎克雷伯菌均为KPC-2,该型是引起肺炎克雷伯菌对碳青酶烯酶耐药的主要机制。
[Abstract]:Objective to investigate the detection methods of Klebsiella pneumoniae (KPC) pneumonia and the mechanism of Klebsiella pneumoniae resistance to carbapenems, the possible therapeutic drugs. Methods 36 strains of Klebsiella pneumoniae resistant to carbapenem pneumonia confirmed by K-B method were selected, and the modified Hodge test and Carba NP test were used. The plant was verified by PCR amplification, agarose gel electrophoresis and gene sequencing of the electrophoresis positive bands. Results the K-B assay showed that 36 strains of Klebsiella pneumoniae resistant to carbenol, followed by Hodge, 33 (91.7%) were KPC pneumonia KK Lei Bojun, and carbenes were resistant to tetracycline and rice. The 33 strains of Hodge test were confirmed by Carba NP test, of which 27 (87.9%) positive and 6 (12.1%) negative, 33 strains of Hodge test positive for PCR amplification and electrophoresis test, 27 of the target bands (340bp) were positive, and the results were in accordance with the Carba NP test results. The 27 strains of Klebsiella pneumoniae appeared in the target band. Then the KPC-2 resistant gene PCR amplified products were sequenced and all were KPC-2 type resistance genes. Conclusion the K-B method of drug sensitivity test plus the subsequent improved Hodge test proved that the KPC pneumonia Klebsiella pneumoniae has a higher coincidence, and the effective screening method for the detection of Klebsiella pneumoniae of KPC pneumonia was found in the.K-B method drug sensitivity test for KPC pneumonia, which was proved to be a substitute for Klebsiella pneumoniae. KPC pneumonia Klebsiella pneumoniae, one of the drugs for effective treatment, was detected by.Hodge test. There was a false positive rate of 12.1% (6/33),.Carba NP test and PCR amplification test, which confirmed the effect of Klebsiella pneumoniae. The result of sequencing showed that all Klebsiella pneumoniae was KPC-2, and this type was The main mechanism of Klebsiella pneumoniae resistance to C - tase is caused by Klebsiella pneumoniae.
【作者单位】：安徽医科大学附属省立医院呼吸内科;安徽医科大学第一附属医院检验科;安徽医科大学附属省立医院感染办;
【基金】：国家自然科学基金(编号:81171618)
【分类号】：R446.5

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