A Single-Laboratory Performance Evaluation of MALDI-TOF MS in Rapid Identification of Staphylococcus aureus, Cronobacter sakazakii, Vibrio parahaemolyticus, and Some Closely Related Bacterial Species of Public Health Importance.

Irshad M Sulaiman, Nancy Miranda, Whitney Hook, Joseph Mendoza, Quincy Kumfert, Tamayo Barnes, Kidon Sung, Saeed Khan, Mohamed Nawaz, Pratik Banerjee, Steven Simpson, Kevin Karem
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Abstract

Background: Staphylococcus is a genus of Gram-positive bacteria, known to cause food poisoning and gastrointestinal illness in humans. Additionally, the emergence of methicillin-resistant S. aureus (MRSA) strains has caused a major health care burden worldwide. Cronobacter is a group of Gram-negative bacteria that can survive in extreme dry conditions. Cronobacter sakazakii is known to contaminate powdered infant formula and cause life-threatening infections in neonates. Vibrio is a genus of human-pathogenic Gram-negative bacteria that can cause foodborne illness by consuming undercooked or raw seafood. Vibrio parahaemolyticus can cause serious gastrointestinal disease in humans. Thus, rapid identification of Staphylococcus spp., Cronobacter spp., and Vibrio spp. is crucial for the source tracking of contaminated food, as well as to measure the transmission dynamics of these bacterial pathogens causing foodborne diseases and outbreaks.

Objective: This single-laboratory performance evaluation study used the VITEK MS system to evaluate the potential of MALDI-TOF MS technology for rapid identification of S. aureus-like, C. sakazakii-like, and V. parahaemolyticus-like isolates of public health importance.

Method: A total of 226 isolates recovered from various food, environmental surveillance samples, and other sources were identified by bioMérieux VITEK 2 and VITEK MS systems as Staphylococcus spp., Cronobacter spp., and Vibrio spp. Five American Type Culture Collection (ATCC) reference Gram-positive and Gram-negative bacterial isolates were also tested to complete the study. In addition, for some Staphylococcus spp. isolates, whole genome sequencing (WGS) and DNA sequencing of 16S rRNA partial region were also performed for species identification.

Results: The VITEK MS system was able to provide species identification to all 96 isolates of Staphylococcus spp. and to all 29 isolates of Vibrio spp. examined with a high confidence value (99.9%). Similarly, species identification was observed for the majority of spots (245 of 303) for the 101 Cronobacter spp. isolates (∼82.0%) with a high confidence value (99.9%), and genus level identification was noticed for the rest of the Cronobacter spp. isolates (18.0%; 58 of the 303 spots) analyzed. Species identification data generated by VITEK 2 system were comparable to data obtained by the VITEK MS system.

Conclusions: The VITEK MS system is a reliable high-throughput platform that can rapidly identify Staphylococcus, Vibrio, and Cronobacter to the genus level, as well as S. aureus, C. sakazakii, V. parahaemolyticus, and other closely related foodborne isolates and bacterial isolates from additional sources, in most cases.

Highlights: The VITEK MS system can be used in the rapid genus and species identification of human-pathogenic Staphylococcus spp., Cronobacter spp., and Vibrio spp. isolates.

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MALDI-TOF MS在快速鉴定金黄色葡萄球菌、阪崎克罗诺杆菌、副溶血性弧菌和一些具有公共卫生重要性的密切相关细菌中的单实验室性能评估。
背景:葡萄球菌是一属革兰氏阳性菌,已知可引起人类食物中毒和胃肠道疾病。此外,耐甲氧西林金黄色葡萄球菌(MRSA)菌株的出现在全球范围内造成了重大的医疗负担。克罗诺杆菌是一组革兰氏阴性细菌,可以在极端干燥的条件下生存。众所周知,阪崎克罗诺杆菌会污染婴儿配方奶粉,并导致新生儿感染危及生命。弧菌是一种人类致病性革兰氏阴性菌,食用未煮熟或生的海鲜会导致食源性疾病。副溶血性弧菌可引起人类严重的胃肠道疾病。因此,快速鉴定葡萄球菌属、克罗诺杆菌属和弧菌属对于追踪受污染食品的来源以及测量这些导致食源性疾病和暴发的细菌病原体的传播动态至关重要。目的:本单实验室性能评估研究使用VITEK MS系统评估MALDI-TOF MS技术快速鉴定具有公共卫生重要性的金黄色葡萄球菌样、阪崎肠杆菌样和副溶血性弧菌样分离株的潜力。方法:通过bioMérieux VITEK 2和VITEK MS系统,从各种食品、环境监测样品和其他来源中回收的226个分离株被鉴定为葡萄球菌属、克罗诺杆菌属和弧菌属。还对5个美国典型培养物保藏中心(ATCC)参考革兰氏阳性和革兰氏阴性菌株进行了检测,以完成研究。此外,对于一些葡萄球菌,还进行了分离物全基因组测序(WGS)和16S rRNA部分区域的DNA测序以进行物种鉴定。结果:VITEK-MS系统能够对所有96个葡萄球菌分离株和所有29个弧菌分离株进行物种鉴定。检测结果具有高置信度值(99.9%)。同样,101个克罗诺杆菌分离株(~82.0%)的大多数斑点(303个斑点中的245个)也观察到物种鉴定,其余Cronobacter spp.分离株(18.0%;303个斑点中的58个)进行了属级鉴定。VITEK 2系统产生的物种鉴定数据与VITEK MS系统获得的数据相当。结论:VITEK-MS系统是一个可靠的高通量平台,在大多数情况下,它可以快速鉴定属水平的葡萄球菌、弧菌和克罗诺杆菌,以及金黄色葡萄球菌、阪崎肠杆菌、副溶血性弧菌和其他密切相关的食源性分离株,以及来自其他来源的细菌分离株。亮点:VITEK-MS系统可用于人类致病性葡萄球菌、克罗诺杆菌和弧菌分离株的快速属种鉴定。
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