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基质辅助激光解吸电离飞行时间质谱(MALDI-TOF MS)鉴定与生化试验的比较:一项针对奶牛场环境中肠杆菌的研究

The Matrix-Assisted Laser Desorption Ionization-Time of Flight Mass Spectrometry (MALDI-TOF MS) identification versus biochemical tests: a study with enterobacteria from a dairy cattle environment.

作者信息

Rodrigues Naiara Miranda Bento, Bronzato Greiciane França, Santiago Gabrielli Stefaninni, Botelho Larissa Alvarenga Batista, Moreira Beatriz Meurer, Coelho Irene da Silva, Souza Miliane Moreira Soares de, Coelho Shana de Mattos de Oliveira

机构信息

Universidade Federal Rural do Rio de Janeiro, Departamento de Microbiologia e Imunologia Veterinária, Seropédica, RJ, Brazil.

Laboratório de Investigação em Microbiologia Médica - LIMM, Instituto de Microbiologia Paulo de Goés, Rio de Janeiro, RJ, Brazil.

出版信息

Braz J Microbiol. 2017 Jan-Mar;48(1):132-138. doi: 10.1016/j.bjm.2016.07.025. Epub 2016 Sep 21.

Abstract

Mastitis adversely affects milk production and in general cows do not regain their full production levels post recovery, leading to considerable economic losses. Moreover the percentage decrease in milk production depends on the specific pathogen that caused the infection and enterobacteria are responsible for this greater reduction. Phenotypic tests are among the currently available methods used worldwide to identify enterobacteria; however they tend to misdiagnose the species despite the multiple tests carried out. On the other hand The Matrix-Assisted Laser Desorption Ionization-Time of Flight Mass Spectrometry (MALDI-TOF MS) technique has been attracting attention for its precise identification of several microorganisms at species level. In the current study, 183 enterobacteria were detected in milk (n=47) and fecal samples (n=94) from cows, and samples from water (n=23) and milk lines (n=19). All these samples were collected from a farm in Rio de Janeiro with the specific purpose of presenting the MALDI-TOF MS technique as an efficient methodology to identify Enterobacteriaceae from bovine environments. The MALDI-TOF MS technique results matched the biochemical test results in 92.9% (170/183) of the enterobacteria species and the gyrB sequencing confirmed 100% of the proteomic technique results. The amino acid decarboxylation test made the most misidentifications and Enterobacter spp. was the most misidentified genus (76.9%, 10/13). These results aim to clarify the current biochemical errors in enterobacteria identification, considering isolates from a bovine environment, and show the importance for more careful readings of phenotypic tests which are often used in veterinary microbiology laboratories.

摘要

乳腺炎会对牛奶产量产生不利影响,一般来说,奶牛康复后无法恢复到之前的全产量水平,从而导致可观的经济损失。此外,牛奶产量下降的百分比取决于引起感染的特定病原体,而肠杆菌是导致产量大幅下降的原因。表型测试是目前全球范围内用于鉴定肠杆菌的方法之一;然而,尽管进行了多项测试,但它们仍容易误诊菌种。另一方面,基质辅助激光解吸电离飞行时间质谱(MALDI-TOF MS)技术因其能在种水平上精确鉴定多种微生物而备受关注。在本研究中,从奶牛的牛奶样本(n = 47)和粪便样本(n = 94)以及水样本(n = 23)和奶管线样本(n = 19)中检测到183株肠杆菌。所有这些样本均采集自里约热内卢的一个农场,其特定目的是展示MALDI-TOF MS技术作为一种从牛环境中鉴定肠杆菌科细菌的有效方法。MALDI-TOF MS技术的结果与92.9%(170/183)的肠杆菌种的生化测试结果相符,gyrB测序证实了蛋白质组学技术结果的100%。氨基酸脱羧酶测试的错误鉴定最多,肠杆菌属是错误鉴定最多的属(76.9%,10/13)。这些结果旨在阐明目前在从牛环境中分离的肠杆菌鉴定中的生化错误,并表明在兽医微生物学实验室中经常使用的表型测试需要更仔细解读的重要性。

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本文引用的文献

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Taxonomic evaluation of the genus Enterobacter based on multilocus sequence analysis (MLSA): proposal to reclassify E. nimipressuralis and E. amnigenus into Lelliottia gen. nov. as Lelliottia nimipressuralis comb. nov. and Lelliottia amnigena comb. nov., respectively, E. gergoviae and E. pyrinus into Pluralibacter gen. nov. as Pluralibacter gergoviae comb. nov. and Pluralibacter pyrinus comb. nov., respectively, E. cowanii, E. radicincitans, E. oryzae and E. arachidis into Kosakonia gen. nov. as Kosakonia cowanii comb. nov., Kosakonia radicincitans comb. nov., Kosakonia oryzae comb. nov. and Kosakonia arachidis comb. nov., respectively, and E. turicensis, E. helveticus and E. pulveris into Cronobacter as Cronobacter zurichensis nom. nov., Cronobacter helveticus comb. nov. and Cronobacter pulveris comb. nov., respectively, and emended description of the genera Enterobacter and Cronobacter.基于多位点序列分析(MLSA)的肠杆菌属分类评估:建议将 E. nimipressuralis 和 E. amnigenus 重新分类为 Lelliottia gen. nov.,分别为 Lelliottia nimipressuralis comb. nov. 和 Lelliottia amnigena comb. nov.,将 E. gergoviae 和 E. pyrinus 重新分类为 Pluralibacter gen. nov.,分别为 Pluralibacter gergoviae comb. nov. 和 Pluralibacter pyrinus comb. nov.,将 E. cowanii、E. radicincitans、E. oryzae 和 E. arachidis 重新分类为 Kosakonia gen. nov.,分别为 Kosakonia cowanii comb. nov.、Kosakonia radicincitans comb. nov.、Kosakonia oryzae comb. nov. 和 Kosakonia arachidis comb. nov.,将 E. turicensis、E. helveticus 和 E. pulveris 重新分类为 Cronobacter 属,分别为 Cronobacter zurichensis nom. nov.、Cronobacter helveticus comb. nov. 和 Cronobacter pulveris comb. nov.,并修订了肠杆菌属和 Cronobacter 属的描述。
Syst Appl Microbiol. 2013 Jul;36(5):309-19. doi: 10.1016/j.syapm.2013.03.005. Epub 2013 Apr 28.

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