双重过滤法联合MALDI-TOF MS快速检测尿液病原菌  

Combination of dual-filtration method with MALDI-TOF MS for rapid detection of urinary pathogens

作　　者：徐金莲[1] 陈学兵[1] 李书琴 肖学会[1] 王沛[1] XU Jinlian;CHEN Xuebing;LI Shuqin;XIAO Xuehui;WANG Pei(The First People′s Hospital of Jingmen, Jingmen 448000, Hubei, China)

机构地区：[1]荆门市第一人民医院检验科,湖北荆门448000

出　　处：《临床检验杂志》2021年第5期346-349,353,共5页Chinese Journal of Clinical Laboratory Science

基　　金：荆门市科技基金项目(2019YFYB029)。

摘　　要：目的采用一种微孔滤膜双重过滤法富集纯化尿液中病原菌,探讨此方法与UF1000i全自动尿沉渣分析仪联合基质辅助激光解吸电离飞行时间质谱(MALDI-TOF MS)在快速检测尿液病原菌中的临床应用。方法收集2020年5—8月门诊及住院疑似尿路感染患者中段尿标本1583份,采用传统细菌定量培养和MALDI-TOF MS鉴定。同时应用微孔滤膜双重过滤法对UF1000i细菌计数≥10^(5)个/mL尿液进行过滤,过滤后的细菌涂抹于靶板上进行MALDI-TOF MS鉴定,与培养法鉴定结果比较分析。结果1583份中段尿标本中,272份标本UF1000i细菌计数阳性且培养菌落数≥10^(4) CFU/mL。与培养法鉴定结果比较,双重过滤法对229份菌落数≥10^(4) CFU/mL的单一菌生长标本鉴定符合率为92.1%(211/229),其中革兰阴性菌符合率为94.7%(177/187),革兰阳性菌符合率为84.6%(33/39),真菌符合率为33.3%(1/3)。双重过滤法对2种菌生长标本仅能鉴定优势菌,43份2种菌混合生长标本优势菌鉴定符合率为72.1%(31/43)。双重过滤法中尿液病原菌富集纯化时间为2~3 min。结论微孔滤膜双重过滤法是一种快速有效的富集纯化尿液病原菌的方法,联合UF1000i和MALDI-TOF MS可实现快速检测尿液中病原菌。Objective To enrich and purify pathogenic bacteria in urine by a dual-filtration method with millipore membrane,and explore the clinical application of UF1000i,dual-filtration combined with matrix-assisted laser desorption ionization time-of-flight mass spectrometry(MALDI-TOF MS)in rapid detection for urinary pathogens.Methods A total of 1583 urine samples of were collected from the patients with suspected urinary tract infection during May 2020 to August 2020.The traditional blood agar plates were used for quantitative culture of bacteria,and MALDI-TOF MS identification was also performed.The urine samples with UF1000i bacterial count≥10^(5)/mL were filtered by dual-filtration method.The filtered bacteria were smeared on the target plate for MALDI-TOF MS identification,the results of which were compared with those of the culture method.Results Among the 1583 middle urine specimens,272 were found to be positive UF1000i bacterial count with colony count≥10^(4) CFU/mL.Compared with the results of culture method,the identification coincidence rate of dual-filtration method was 92.1%(211/229)for a single growth specimen with≥10^(4) CFU/mL of cultured colonies,of which gram-negative bacteria,gram-positive bacteria and fungi was 94.7%(177/187),84.6%(33/39)and 33.3%(1/3),respectively.The dual-filtration method could only identify the dominant bacteria in the specimens with two kinds of bacteria growing,and the identification coincidence rate of dominant bacteria was 72.1%(31/43)in 43 specimens with two kinds of mixed growing bacteria.The enrichment and purification of urine in the dual-filtration method just needed 2 to 3 minutes for each sample.Conclusion Dual-filtration method with millipore membrane should be a rapid and effective method for enrichment and purification of pathogens in urine.Combination of UF1000i and MALDI-TOF MS may speed up the detection process for pathogens in urine.

关 键 词：双重过滤法 基质辅助激光解吸电离飞行时间质谱 尿路感染 病原菌 快速检测 

分 类 号：R446.5[医药卫生—诊断学]