检验医学 ›› 2024, Vol. 39 ›› Issue (9): 880-887.DOI: 10.3969/j.issn.1673-8640.2024.09.011

• 论著 • 上一篇    下一篇

不同类型临床样本来源肺炎克雷伯菌耐药性和毒力分析

郭超楠1, 王妍妍1, 张贝1, 庞敬莹2, 崔非非2, 赵永新1, 苏兵1   

  1. 1.新乡医学院第三附属医院检验科,河南 新乡 453003
    2.新乡医学院第一附属医院检验科,河南 新乡 453003
  • 收稿日期:2023-07-14 修回日期:2024-02-01 出版日期:2024-09-30 发布日期:2024-10-15
  • 作者简介:郭超楠,女,1992年生,硕士,主管检验师,主要从事临床病原微生物检验工作。

Analysis of drug resistance and virulence of Klebsiella pneumoniae from different clinical samples

GUO Chaonan1, WANG Yanyan1, ZHANG Bei1, PANG Jingying2, CUI Feifei2, ZHAO Yongxin1, SU Bing1   

  1. 1. Department of Clinical Laboratory,the Third Affiliated Hospital of Xinxiang Medical University,Xinxiang 453003,Henan,China
    2. Department of Clinical Laboratory,the First Affiliated Hospital of Xinxiang Medical University,Xinxiang 453003,Henan,China
  • Received:2023-07-14 Revised:2024-02-01 Online:2024-09-30 Published:2024-10-15

摘要:

目的 分析不同类型临床样本来源肺炎克雷伯菌(KP)的耐药性、生物膜形成能力、毒力基因和耐药基因的携带特点,及其相互间的联系,为控制医院内感染提供参考。方法 收集2021年11月—2022年9月新乡市3家教学医院分离自痰液、尿液(中段尿)和血液样本的148株KP。采用全自动微生物鉴定系统进行细菌鉴定和药物敏感性分析,采用聚合酶链反应(PCR)检测菌株荚膜血清型、毒力基因和耐药基因,采用拉丝试验检测菌株高黏液表型,采用结晶紫染色法检测菌株生物膜的形成情况。结果 148株KP中,有55株(37.2%)分离自痰液样本,40株(27.0%)分离自血液样本,53株(35.8%)分离自尿液样本。尿液样本的强生物膜形成菌株分离率 (54.3%)高于痰液样本(27.5%)和血液样本(29.7%),差异有统计学意义(P=0.023);耐药率(69.8%)高于分离自痰液样本的菌株(49.1%)(P=0.028)。携带率(85.8%)最高的耐药基因是blaSHV-1。分离自痰液样本的KP毒力基因iroNrmpA检出率高于尿液样本(P=0.004)和血液样本(P=0.012);wabG fimHmrkD基因携带率均>93.2%。携带iroNiucArmpA基因和K1、K2血清型的KP对抗菌药物较敏感,且易检出高黏液表型(P<0.05)。高黏液表型KP菌株同时携带4种及以上毒力基因的构成比高于非高黏液表型KP菌株(P<0.001),且对抗菌药物更敏感(P<0.05)。74.5%的痰液样本来源KP同时携带4种及以上毒力基因,高于血液样本来源(52.5%)和尿液样本来源(49.1%)的KP(P=0.016);形成生物膜的KP菌株wabG mrkDugefimH基因携带率更高(P<0.05)。结论 尿液样本来源的KP耐药率较高,且易形成强生物膜。某些特定毒力基因普遍存在于KP临床分离株中。痰液样本来源的KP毒力基因构成复杂,iroNrmpA基因与呼吸系统KP感染有一定关系。iroNiucArmpA基因和K1、K2血清型与KP高黏液表型有关,且携带此类基因和血清型的KP菌株对抗菌药物更敏感。

关键词: 肺炎克雷伯菌, 痰液, 中段尿, 血液, 高黏液表型, 毒力基因, 生物膜, 耐药

Abstract:

Objective To analyze the characteristics of drug resistance,biofilm forming ability,virulence gene and drug resistance gene in Klebsiella pneumoniae (KP) from different clinical samples,and to provide a reference for controlling nosocomial infection. Methods Totally,148 KP isolates from sputum,midstream urine and blood samples were collected from 3 teaching hospitals in Xinxiang from November 2021 to September 2022. Bacterial identification and drug susceptibility test were performed by automatic microbial identification system. Polymerase chain reaction (PCR) was used to determine serotypes,virulence genes and drug resistance genes. Stringing test was used to determine high viscosity phenotype,and crystal violet staining was used to determine biofilm formation. Results Among 148 KP isolates,55(37.2%)isolates were isolated from sputum samples,40(27.0%)isolates were isolated from blood samples,and 53(35.8%)isolates were isolated from midstream urine samples. The isolation rate of strong biofilm forming isolates in midstream urine samples (54.3%) was higher than those in sputum samples(27.45%)and in blood samples(29.73%)(P=0.027),and the drug resistance rate (69.8%) was higher than that in sputum samples (49.1%)(P=0.028). The highest carrying rate of drug resistance gene was blaSHV-1(85.8%). The determination rates of KP virulence genes,iroN and rmpA,in sputum samples were higher than those in midstream urine samples (P=0.004) and blood samples (P=0.012). The carrying rates of wabGfimH and mrkD genes were >93.2%. KP carrying iroNiucArmpA genes and K1 and K2 serotypes were sensitive to antibiotics and easily determined high viscosity phenotype (P<0.05). The proportion of KP isolates with high viscosity phenotype was higher than that of KP isolates without high viscosity phenotype (P<0.001),and KP isolates with high viscosity were more sensitive to antibiotics (P<0.05). The 74.5% of sputum samples had KP with 4 or more virulence genes,which was higher than those of blood samples (52.5%) and midstream urine samples (49.1%) (P=0.016). The biofilm forming KP isolates carried more wabG mrkDuge and fimH genes (P<0.05). Conclusions he drug resistance rate of KP from midstream urine samples is high,which is easy to form strong biofilm. Some specific virulence genes are prevalent in KP. The composition of KP virulence genes from sputum samples is complex. The iroN and rmpA genes are associated with KP infection in respiratory system. The iroNiucArmpA genes and K1,K2 serotypes are associated with high viscosity phenotypes,and KP isolates containing these genes and serotypes are more sensitive to antibiotics.

Key words: Klebsiella pneumoniae, Sputum, Midstream urine, Blood, High viscosity, Virulence gene, Biofilm, Drug resistance

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