Laser Optical Sensor, a Label-Free On-Plate <named-content content-type="genus-species">Salmonella enterica</named-content> Colony Detection Tool

ABSTRACT We investigated the application capabilities of a laser optical sensor, BARDOT (bacterial rapid detection using optical scatter technology) to generate differentiating scatter patterns for the 20 most frequently reported serovars of Salmonella enterica. Initially, the study tested the class...

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Autores principales:	Atul K. Singh, Amanda M. Bettasso, Euiwon Bae, Bartek Rajwa, Murat M. Dundar, Mark D. Forster, Lixia Liu, Brent Barrett, Judith Lovchik, J. Paul Robinson, E. Daniel Hirleman, Arun K. Bhunia
Formato:	article
Lenguaje:	EN
Publicado:	American Society for Microbiology 2014
Materias:	Microbiology QR1-502
Acceso en línea:	https://doaj.org/article/17437b611d1b463eaa418f8b5aaa51d5
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spelling	oai:doaj.org-article:17437b611d1b463eaa418f8b5aaa51d52021-11-15T15:45:10ZLaser Optical Sensor, a Label-Free On-Plate <named-content content-type="genus-species">Salmonella enterica</named-content> Colony Detection Tool10.1128/mBio.01019-132150-7511https://doaj.org/article/17437b611d1b463eaa418f8b5aaa51d52014-02-01T00:00:00Zhttps://journals.asm.org/doi/10.1128/mBio.01019-13https://doaj.org/toc/2150-7511ABSTRACT We investigated the application capabilities of a laser optical sensor, BARDOT (bacterial rapid detection using optical scatter technology) to generate differentiating scatter patterns for the 20 most frequently reported serovars of Salmonella enterica. Initially, the study tested the classification ability of BARDOT by using six Salmonella serovars grown on brain heart infusion, brilliant green, xylose lysine deoxycholate, and xylose lysine tergitol 4 (XLT4) agar plates. Highly accurate discrimination (95.9%) was obtained by using scatter signatures collected from colonies grown on XLT4. Further verification used a total of 36 serovars (the top 20 plus 16) comprising 123 strains with classification precision levels of 88 to 100%. The similarities between the optical phenotypes of strains analyzed by BARDOT were in general agreement with the genotypes analyzed by pulsed-field gel electrophoresis (PFGE). BARDOT was evaluated for the real-time detection and identification of Salmonella colonies grown from inoculated (1.2 × 102 CFU/30 g) peanut butter, chicken breast, and spinach or from naturally contaminated meat. After a sequential enrichment in buffered peptone water and modified Rappaport Vassiliadis broth for 4 h each, followed by growth on XLT4 (~16 h), BARDOT detected S. Typhimurium with 84% accuracy in 24 h, returning results comparable to those of the USDA Food Safety and Inspection Service method, which requires ~72 h. BARDOT also detected Salmonella (90 to 100% accuracy) in the presence of background microbiota from naturally contaminated meat, verified by 16S rRNA sequencing and PFGE. Prolonged residence (28 days) of Salmonella in peanut butter did not affect the bacterial ability to form colonies with consistent optical phenotypes. This study shows BARDOT’s potential for nondestructive and high-throughput detection of Salmonella in food samples. IMPORTANCE High-throughput screening of food products for pathogens would have a significant impact on the reduction of food-borne hazards. A laser optical sensor was developed to screen pathogen colonies on an agar plate instantly without damaging the colonies; this method aids in early pathogen detection by the classical microbiological culture-based method. Here we demonstrate that this sensor was able to detect the 36 Salmonella serovars tested, including the top 20 serovars, and to identify isolates of the top 8 Salmonella serovars. Furthermore, it can detect Salmonella in food samples in the presence of background microbiota in 24 h, whereas the standard USDA Food Safety and Inspection Service method requires about 72 h.Atul K. SinghAmanda M. BettassoEuiwon BaeBartek RajwaMurat M. DundarMark D. ForsterLixia LiuBrent BarrettJudith LovchikJ. Paul RobinsonE. Daniel HirlemanArun K. BhuniaAmerican Society for MicrobiologyarticleMicrobiologyQR1-502ENmBio, Vol 5, Iss 1 (2014)
institution	DOAJ
collection	DOAJ
language	EN
topic	Microbiology QR1-502
spellingShingle	Microbiology QR1-502 Atul K. Singh Amanda M. Bettasso Euiwon Bae Bartek Rajwa Murat M. Dundar Mark D. Forster Lixia Liu Brent Barrett Judith Lovchik J. Paul Robinson E. Daniel Hirleman Arun K. Bhunia Laser Optical Sensor, a Label-Free On-Plate <named-content content-type="genus-species">Salmonella enterica</named-content> Colony Detection Tool
description	ABSTRACT We investigated the application capabilities of a laser optical sensor, BARDOT (bacterial rapid detection using optical scatter technology) to generate differentiating scatter patterns for the 20 most frequently reported serovars of Salmonella enterica. Initially, the study tested the classification ability of BARDOT by using six Salmonella serovars grown on brain heart infusion, brilliant green, xylose lysine deoxycholate, and xylose lysine tergitol 4 (XLT4) agar plates. Highly accurate discrimination (95.9%) was obtained by using scatter signatures collected from colonies grown on XLT4. Further verification used a total of 36 serovars (the top 20 plus 16) comprising 123 strains with classification precision levels of 88 to 100%. The similarities between the optical phenotypes of strains analyzed by BARDOT were in general agreement with the genotypes analyzed by pulsed-field gel electrophoresis (PFGE). BARDOT was evaluated for the real-time detection and identification of Salmonella colonies grown from inoculated (1.2 × 102 CFU/30 g) peanut butter, chicken breast, and spinach or from naturally contaminated meat. After a sequential enrichment in buffered peptone water and modified Rappaport Vassiliadis broth for 4 h each, followed by growth on XLT4 (~16 h), BARDOT detected S. Typhimurium with 84% accuracy in 24 h, returning results comparable to those of the USDA Food Safety and Inspection Service method, which requires ~72 h. BARDOT also detected Salmonella (90 to 100% accuracy) in the presence of background microbiota from naturally contaminated meat, verified by 16S rRNA sequencing and PFGE. Prolonged residence (28 days) of Salmonella in peanut butter did not affect the bacterial ability to form colonies with consistent optical phenotypes. This study shows BARDOT’s potential for nondestructive and high-throughput detection of Salmonella in food samples. IMPORTANCE High-throughput screening of food products for pathogens would have a significant impact on the reduction of food-borne hazards. A laser optical sensor was developed to screen pathogen colonies on an agar plate instantly without damaging the colonies; this method aids in early pathogen detection by the classical microbiological culture-based method. Here we demonstrate that this sensor was able to detect the 36 Salmonella serovars tested, including the top 20 serovars, and to identify isolates of the top 8 Salmonella serovars. Furthermore, it can detect Salmonella in food samples in the presence of background microbiota in 24 h, whereas the standard USDA Food Safety and Inspection Service method requires about 72 h.
format	article
author	Atul K. Singh Amanda M. Bettasso Euiwon Bae Bartek Rajwa Murat M. Dundar Mark D. Forster Lixia Liu Brent Barrett Judith Lovchik J. Paul Robinson E. Daniel Hirleman Arun K. Bhunia
author_facet	Atul K. Singh Amanda M. Bettasso Euiwon Bae Bartek Rajwa Murat M. Dundar Mark D. Forster Lixia Liu Brent Barrett Judith Lovchik J. Paul Robinson E. Daniel Hirleman Arun K. Bhunia
author_sort	Atul K. Singh
title	Laser Optical Sensor, a Label-Free On-Plate <named-content content-type="genus-species">Salmonella enterica</named-content> Colony Detection Tool
title_short	Laser Optical Sensor, a Label-Free On-Plate <named-content content-type="genus-species">Salmonella enterica</named-content> Colony Detection Tool
title_full	Laser Optical Sensor, a Label-Free On-Plate <named-content content-type="genus-species">Salmonella enterica</named-content> Colony Detection Tool
title_fullStr	Laser Optical Sensor, a Label-Free On-Plate <named-content content-type="genus-species">Salmonella enterica</named-content> Colony Detection Tool
title_full_unstemmed	Laser Optical Sensor, a Label-Free On-Plate <named-content content-type="genus-species">Salmonella enterica</named-content> Colony Detection Tool
title_sort	laser optical sensor, a label-free on-plate <named-content content-type="genus-species">salmonella enterica</named-content> colony detection tool
publisher	American Society for Microbiology
publishDate	2014
url	https://doaj.org/article/17437b611d1b463eaa418f8b5aaa51d5
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Laser Optical Sensor, a Label-Free On-Plate <named-content content-type="genus-species">Salmonella enterica</named-content> Colony Detection Tool

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