| 张才庆,李艳,吴青.基质辅助激光解吸电离飞行时间质谱技术直接鉴定尿路感染病原菌[J].安徽医药,2019,23(11):2151-2155. |
| 基质辅助激光解吸电离飞行时间质谱技术直接鉴定尿路感染病原菌 |
| Direct identification of urinary tract infection pathogens by matrix?assisted laser desorption/ionization time?of?flight mass spectrometry |
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| DOI：10.3969/j.issn.1009?6469.2019.11.008 |
| 中文关键词: 光谱法，质量，基质辅助激光解吸电离 集落计数，微生物 尿路感染 诊断 |
| 英文关键词: Spectrometry,mass,matrix?assisted laser desorption?ionization Colony count,microbial Urinary tract infection Diagnosis |
| 基金项目:国家自然科学基金面上项目(81772265) |
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| 中文摘要: |
| 目的利用基质辅助激光解吸电离飞行时间质谱技术( MALDI?TOF MS)建立尿路感染病原菌快速、准确的鉴定方法。方法收集 2017年 6月至 2018年 11月武汉大学人民医院中段尿培养标本 4 493例，采用差速离心法富集细菌，利用 MALDI? TOF MS对富集后的菌液进行快速鉴定，并与传统培养 1~2 d后形成的菌落最终鉴定结果进行比较，同时将尿路感染常见病原菌作倍比稀释评估细菌被鉴定的最低检测限度。结果 4 493例中段尿标本中， 775例单一菌感染且细菌数 ≥105cfu/mL的样本运用 MALDI?TOF MS直接检测出 663株菌，尿病原菌直接质谱鉴定率为 85.55%(663/775)菌株的属、种鉴定符合率分别为 85.55%(663/775)和 83.10%(644/775)；其中 557株革兰阴性菌属、种鉴定符合率分别为 94.80%(，528/557)和 94.08%(524/557)， 159株革兰阳性菌属、种鉴定符合率分别为 71.07%(113/159)和 64.78%(103/159)59例真菌属、种鉴定符合率分别为 37.29%(22/59)和 28.81%(17/59)。 183例样本为 2种菌混合感染，采用 MALDI?TOF MS直接，鉴定出 136例样本中的 1种优势菌，直接质谱鉴定率为 74.32%(136/183)，同时发现不同浓度的细菌计数与 MALDI?TOF MS阳性率有一定的关系，在 1.5×104cfu/mL革兰阴性菌比革兰阳性菌的质谱鉴定率高。结论当尿液中的细菌浓度达到一定的检测限度时， MALDI?TOF MS可以直接检测其细菌，且检出率高、结果可靠； MALDI?TOF MS对混合菌感染的样本也有一定鉴定能力，该方法能够极大地缩短病原菌的鉴定时间，为临床诊断提供有效、准确、可靠的手段。 |
| 英文摘要: |
| Objective To establish a rapid and accurate method of urinary tract infection pathogens by matrix?assisted laser desorp?tion/ionization time?of?flight mass spectrometry.Methods A total of 4 493 clean urine samples were collected from June 2017 toNovember 2018.The bacteria were enriched by differential centrifugation method，and the enriched bacterial liquid was quickly identified by MALDI?TOF MS，and compared with the final identification results of bacterial colonies formed after 1?2 d of tradition? al culture.At the same time，the minimal detection limit for the identification of common pathogens of urinary tract infection was de?termined by multiple dilution.Results Among the 4 493 cases of midcourse urine specimens，775 cases of single bacterial infec? tion with a bacterial count of more than 105cfu/ml were directly detected by MALDI?TOF MS.The identification rate of urinary pathogens was 85.55%(663/775)by direct mass spectrometry，and the identification rate of genus and species were 85.55%(663/ 775)and 83.10%(644/775)，respectively.Among them，the identification coincidence rates of 557 gram?negative bacteria genera and species were 94.80%(528/557)and 94.08%(524/557)，159 gram?positive bacteria genera and species were 71.07%(113/ 159)and 64.78%(103/159)，and 59 fungi genera and species were 37.29%(22/59)and 28.81%(17/59)，respectively.The 183 samples were mixed infection of 2 kinds of bacteria，and 1 dominant bacteria in 136 samples was directly identified by MALDI? TOF MS，and the direct mass spectrometry identification rate was 74.32%(136/183).At the same time，it was found that there was a certain relationship between the bacterial count of different concentrations and MALDI?TOF MS positive，and the mass spectrome? try identification rate of gram?negative bacteria in 1.5×104cfu/ml was higher than that of gram?positive bacteria.Conclusion when the concentration of bacteria in urine reaches a certain limit，MALDI?TOF MS can directly detect the bacteria，with high detectionrate and reliable results.MALDI?TOF MS also has a certain ability to identify samples infected with mixed bacteria.This method cangreatly shorten the identification time of pathogenic bacteria and provide effective，accurate and reliable means for clinical diagnosis. |
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