| Development and Validation of a Real-Time TaqMan Assay for the Detection and Enumeration of Pseudomonas fluorescens ATCC 13525 Used as a Challenge Organism in Testing of Food Equipments. | |
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MedLine Citation:
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PMID: 22250861 Owner: NLM Status: Publisher |
Abstract/OtherAbstract:
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Pseudomonas fluorescens ATCC 13525 is used as the challenge organism to evaluate the efficacy of the clean-in-place (CIP) process of food equipment (automatic ice-maker) as per NSF/ANSI Standard 12. Traditional culturing methodology is presently used to determine the concentration of the challenge organism, which takes 48 h to confirm the cell density. Storage of the challenge preparation in the refrigerator might alter the cell density as P. fluorescens is capable of growing at 4 °C. Also, background organism can grow on the Pseudomonas F agar (PFA) used for the recovery of P. fluorescens thus affecting the results of the test. Real-time TaqMan assay targeting the cpn60 gene was developed for the enumeration and the identification of P. fluorescens because of its specificity, accuracy, and shorter turnaround time. The TaqMan primer-probe pair developed using the Allele ID® 7.0 probe design software was highly specific and sensitive for the target organism. The sensitivity of the assay was 10 colony forming units (CFU)/mL. The assay was also successful in determining the concentration of the challenge preparation within 2 h. Based on these observations, TaqMan assay targeting the cpn60 gene can be efficiently used for strain level identification and enumeration of bacteria. Practical Application: Pseudomonas fluorescens ATCC 13525 is used as a challenge organism in the efficacy testing of clean-in-place process of food equipments. Currently, culturing technique is used for its identification and estimation, which is not only time-consuming but also prone to error. Real-time TaqMan assay is more specific, sensitive, and accurate along with a shorter turnaround time compared to culturing techniques, thereby increasing the overall quality of the testing methodology to evaluate the clean-in-place process critical for the food industry to protect public health and safety. |
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Authors:
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Ratul Saha; Lorelle L Bestervelt; Robert S Donofrio |
Publication Detail:
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Type: JOURNAL ARTICLE Date: 2012-1-17 |
Journal Detail:
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Title: Journal of food science Volume: - ISSN: 1750-3841 ISO Abbreviation: - Publication Date: 2012 Jan |
Date Detail:
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Created Date: 2012-1-18 Completed Date: - Revised Date: - |
Medline Journal Info:
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Nlm Unique ID: 0014052 Medline TA: J Food Sci Country: - |
Other Details:
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Languages: ENG Pagination: - Citation Subset: - |
Copyright Information:
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© 2012 Institute of Food Technologists® |
Affiliation:
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Authors are with Dept. of Microbiology and Molecular Biology, NSF Intl., 789 N Dixboro Rd., Ann Arbor, MI 48105, U.S.A. Direct inquiries to author Saha (E-mail: rsaha@nsf.org). |
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From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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