高毒力肺炎克雷伯菌临床感染特征及质谱同源性分析

Homology analysis and clinical infection characteristics of hypervirulent Klebsiella pneumonia

目的从敏感和多重耐药的肺炎克雷伯菌中筛选出hvKP表型株,对比探讨其临床感染特征的分布和同源性。方法回顾性收集2014至2018年中山大学附属第三医院临床感染标本所分离的敏感和多重耐药的肺炎克雷伯菌共158株,通过黏液拉丝试验(string test,ST)初筛hvKP感染情况,共筛选到65例患者,男性51例,女性14例,平均年龄56岁。构成比分析其临床感染相关信息,基质辅助激光解析电离飞行时间质谱技术(MALDI-TOF MS)对筛选出的hvKP进行种属鉴定及建立新数据库,利用分析软件对所有符合实验条件的菌株进行生物树聚类分析和主成分分析,获得蛋白水平的同源性。结果根据ST试验,158株肺炎克雷伯菌株中,65例患者共筛选到hvKP 65株,其中62株为全敏感菌株,3株为多重耐药菌株,感染者入院主要疾病诊断集中在肝胆疾病、脑病及肝脓肿;人群基础病主要为糖尿病与高血压。聚类分析将hvKP在距离水平线550处分为3群;主成分分析发现部分菌株具有较近亲缘关系,其中占比最大的Kc群主要与肝胆管疾病有关。Ka群主要与手术后感染有关。结论hvKP具有高侵袭力,临床感染表现多样,感染病患多为免疫力低下老年患者。MALDI-TOF MS质谱快速同源性分析对hvKP感染快速筛查具有重要临床意义。

Objective The hvKP phenotype strains were screened from the sensitive and multi-drug resistant Klebsiella pneumoniae,and the distribution and homology of their clinical infection characteristics were compared.Methods A total of 158 Klebsiella pneumoniae strains isolated from clinical infection specimens in the Third Affiliated Hospital of Sun Yat-sen University from 2014 to 2018 were retrospectively collected.The string test(ST)was used to screen the hvKP infection,and a total of 65 were screened.The number of patients included 51 males and 14 females,with an average age of 56 years.The composition ratio of Klebsiellapneumoniae was analyzed for clinical infection related information.Matrix assisted laser desorption-ionization time of flight mass spectrometry(MALDI-TOF MS)was used to identify the hypervirulent Klebsiella pneumoniae and establish a new database,and bio-tree cluster analysis and principal component analysis were carried out for all strains that met the experimental conditions by using analysis software and the homology of protein level was obtained.Results A total of 65 strains hvKP were screened,62 of which were susceptible strains and 3 were multi-drug resistant strains.The main diseases of infected patients were diagnosed as hepatobiliary disease,encephalopathy and liver abscess.The basic diseases of the population were mainly diabetes and hypertension.Dendrogram cluster analysis divided hvKP into 3 groups at a distance of 550 from the horizontal line;principal component analysis found that some strains were closely related,and the Kc group with the largest proportion was mainly related to hepatobiliary diseases.Ka group was mainly related to infection after surgery.Conclusions HvKP was highly invasive and the clinical manifestations of infection were diverse.The infected patients were mostly elderly patients with weakened immunity.The rapid homology analysis of MALDI-TOF MS mass spectrometry has important clinical significance for the rapid screening of hvKP infection.