PDF(Pubmed)
Abstract：
UNASSIGNED: Bacterial foodborne pathogens pose a substantial global public health concern, prompting government agencies and public health organizations to establish food safety guidelines and regulations aimed at mitigating the risk of foodborne illness. The advent of DNA-based amplification coupled with mass spectrometry, known as MassARRAY analysis, has proven to be a highly precise, sensitive, high-throughput, and cost-effective method for bacterial detection. This study aimed to develop, validate, and evaluate a MassARRAY-based assay for the detection and identification of significant enteropathogenic bacteria.
UNASSIGNED: The MassARRAY-based assay was developed for the detection of 10 crucial bacterial foodborne pathogens, including Campylobacter coli, Campylobacter jejuni, Clostridium perfringens, Escherichia coli, Enterococcus faecalis, Enterococcus faecium, Listeria monocytogenes, Salmonella spp., Shigella spp., and Staphylococcus aureus. The assay was optimized using the reference gDNA (n = 19), followed by validation using gDNA (n = 85) of reference and laboratory isolates. Additionally, the evaluation of the assay\'s reaction using a mixture of gDNA from all nine targeted species was performed. The limit of detection of the developed MassARRAY-based assay was determined using bacterial cells. Moreover, the validation method for field samples was evaluated by comparing it with standard microbiological testing methods routinely analyzed.
UNASSIGNED: The developed MassARRAY-based assay demonstrated 100% concordance with known bacterial pure cultures. The assay\'s reaction using a mixture of gDNA from all nine targeted species revealed the MassARRAY\'s capability to detect all targeted species in a single assay with the lowest concentration of 1 ng/μL of gDNA. The limits of detection of the assay range from 357 ± 101 to 282,000 ± 79,196 cells. Moreover, the validation of the assay in field samples revealed a 100% correlation between the data obtained from the standard microbiological method and the MassARRAY-based assay.
UNASSIGNED: These findings suggested that the developed MassARRAY-based assay exhibited the excellence in high-throughput detection of foodborne bacterial pathogens with high accuracy, reliability, and potential applicability within real-world field samples.
UNASSIGNED: The MassARRAY-based assay was developed for the detection of 10 crucial bacterial foodborne pathogens, including Campylobacter coli, Campylobacter jejuni, Clostridium perfringens, Escherichia coli, Enterococcus faecalis, Enterococcus faecium, Listeria monocytogenes, Salmonella spp., Shigella spp., and Staphylococcus aureus. The assay was optimized using the reference gDNA (n = 19), followed by validation using gDNA (n = 85) of reference and laboratory isolates. Additionally, the evaluation of the assay\'s reaction using a mixture of gDNA from all nine targeted species was performed. The limit of detection of the developed MassARRAY-based assay was determined using bacterial cells. Moreover, the validation method for field samples was evaluated by comparing it with standard microbiological testing methods routinely analyzed.
UNASSIGNED: The developed MassARRAY-based assay demonstrated 100% concordance with known bacterial pure cultures. The assay\'s reaction using a mixture of gDNA from all nine targeted species revealed the MassARRAY\'s capability to detect all targeted species in a single assay with the lowest concentration of 1 ng/μL of gDNA. The limits of detection of the assay range from 357 ± 101 to 282,000 ± 79,196 cells. Moreover, the validation of the assay in field samples revealed a 100% correlation between the data obtained from the standard microbiological method and the MassARRAY-based assay.
UNASSIGNED: These findings suggested that the developed MassARRAY-based assay exhibited the excellence in high-throughput detection of foodborne bacterial pathogens with high accuracy, reliability, and potential applicability within real-world field samples.
摘要：
细菌食源性病原体构成了全球公共卫生问题，促使政府机构和公共卫生组织制定旨在降低食源性疾病风险的食品安全指南和法规。基于DNA的扩增与质谱联用的出现，称为MassARRAY分析，已经证明是一个高度精确的，敏感,高通量，和具有成本效益的细菌检测方法。这项研究旨在发展，验证,并评估基于MassARRAY的检测和鉴定重要肠致病菌的方法。
■开发了基于MassARRAY的检测方法，用于检测10种关键的细菌性食源性病原体，包括弯曲杆菌,空肠弯曲杆菌,产气荚膜梭菌,大肠杆菌,粪肠球菌,屎肠球菌,单核细胞增生李斯特菌,沙门氏菌属。,志贺氏菌属。,和金黄色葡萄球菌。使用参考gDNA(n=19)优化测定，然后使用参考和实验室分离株的gDNA（n=85）进行验证。此外,使用来自所有9种目标物种的gDNA混合物对测定反应进行评估。使用细菌细胞确定开发的基于MassARRAY的测定的检测限。此外，通过与常规分析的标准微生物检测方法进行比较,对现场样品的验证方法进行了评价.
■开发的基于MassARRAY的测定证明与已知的细菌纯培养物100%一致。使用来自所有9种目标物种的gDNA混合物进行的测定反应显示，MassARRAY能够在最低浓度为1ng/μL的gDNA的单次测定中检测所有目标物种。该测定的检测限范围为357±101至282,000±79,196个细胞。此外，在现场样本中进行的试验验证显示,从标准微生物学方法获得的数据与基于MassARRAY的试验数据之间存在100%的相关性.
■这些发现表明，开发的基于MassARRAY的检测方法在高通量检测食源性细菌病原体方面表现出卓越的准确性，可靠性,以及在现实世界现场样本中的潜在适用性。
■开发了基于MassARRAY的检测方法，用于检测10种关键的细菌性食源性病原体，包括弯曲杆菌,空肠弯曲杆菌,产气荚膜梭菌,大肠杆菌,粪肠球菌,屎肠球菌,单核细胞增生李斯特菌,沙门氏菌属。,志贺氏菌属。,和金黄色葡萄球菌。使用参考gDNA(n=19)优化测定，然后使用参考和实验室分离株的gDNA（n=85）进行验证。此外,使用来自所有9种目标物种的gDNA混合物对测定反应进行评估。使用细菌细胞确定开发的基于MassARRAY的测定的检测限。此外，通过与常规分析的标准微生物检测方法进行比较,对现场样品的验证方法进行了评价.
■开发的基于MassARRAY的测定证明与已知的细菌纯培养物100%一致。使用来自所有9种目标物种的gDNA混合物进行的测定反应显示，MassARRAY能够在最低浓度为1ng/μL的gDNA的单次测定中检测所有目标物种。该测定的检测限范围为357±101至282,000±79,196个细胞。此外，在现场样本中进行的试验验证显示,从标准微生物学方法获得的数据与基于MassARRAY的试验数据之间存在100%的相关性.
■这些发现表明，开发的基于MassARRAY的检测方法在高通量检测食源性细菌病原体方面表现出卓越的准确性，可靠性,以及在现实世界现场样本中的潜在适用性。
