Ojasanya Rasaq A, Gardner Ian A, Groman David, Saksida Sonja, Saab Matthew E, Thakur Krishna K
Department of Health Management, Atlantic Veterinary College, University of Prince Edward Island, Charlottetown, PE, Canada.
Aquatic Diagnostic Services, Atlantic Veterinary College, University of Prince Edward Island, Charlottetown, PE, Canada.
Front Vet Sci. 2022 Oct 20;9:1031373. doi: 10.3389/fvets.2022.1031373. eCollection 2022.
Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) allows rapid and reliable identification of microorganisms. The accuracy of bacterial identification using MALDI-TOF MS depends on main spectral profiles (MSPs) provided in a quality-assured commercial reference library, which requires ongoing improvement. This study aimed to develop and validate an in-house MALDI-TOF MS MSP to rapidly identify isolated from Atlantic salmon (). The novel MSP was prepared using an isolate of recovered from Atlantic salmon and confirmed by 16S rRNA gene sequencing. Subsequently, a validation set which comprises 29 isolates of were examined from three fishes: Atlantic salmon () ( = 26), American eel () ( = 1), and Atlantic cod () ( = 2). These isolates were randomly selected from the Atlantic Veterinary College, Aquatic Diagnostic Services Bacteriology Laboratory's culture collection to validate the novel MSP. Analytical sensitivity of MALDI-TOF MS using the novel MSP to identify the validation set was 86.2%. Repeatability was assessed by acquiring spectra from 30 different spots of a randomly-selected isolate of , and analyzed spectra from each spot were compared against the novel MSP. The coefficient of variation was 3.3%. The novel MSP clustered with Bruker MSPs ( = 3) of in the reference library and did not falsely identify any closely related bacteria to . This study reports the development of a novel MSP of high analytical sensitivity and specificity for rapid identification of using MALDI-TOF MS.
基质辅助激光解吸/电离飞行时间质谱（MALDI-TOF MS）可实现对微生物的快速可靠鉴定。使用MALDI-TOF MS进行细菌鉴定的准确性取决于质量有保证的商业参考库中提供的主要光谱图谱（MSP），而这需要不断改进。本研究旨在开发并验证一种用于快速鉴定从大西洋鲑鱼中分离出的[具体细菌名称未给出]的内部MALDI-TOF MS MSP。该新型MSP是使用从大西洋鲑鱼中分离出的[具体细菌名称未给出]菌株制备的，并通过16S rRNA基因测序进行了确认。随后，对来自三种鱼类的29株[具体细菌名称未给出]分离株进行了验证：大西洋鲑鱼（[具体数量未给出]）、美洲鳗鲡（[具体数量未给出]）和大西洋鳕鱼（[具体数量未给出]）。这些分离株是从大西洋兽医学院水生诊断服务细菌学实验室的培养物保藏中心随机选取的，以验证该新型MSP。使用新型MSP的MALDI-TOF MS对验证集进行鉴定的分析灵敏度为86.2%。通过从随机选择的一株[具体细菌名称未给出]的30个不同斑点获取光谱来评估重复性，并将每个斑点的分析光谱与新型MSP进行比较。变异系数为3.3%。该新型MSP与参考库中[具体细菌名称未给出]的布鲁克MSP（[具体数量未给出]）聚类，且未错误鉴定任何与[具体细菌名称未给出]密切相关的细菌。本研究报告了一种用于使用MALDI-TOF MS快速鉴定[具体细菌名称未给出]的具有高分析灵敏度和特异性的新型MSP的开发情况。
