Utilization of Fluorogenic Assay for Rapid Detection of Escherichia coli in Acidic Fruit Juice
Abstract:This study was undertaken to investigate interference by acids commonly found in fruit juice in Escherichia coli assays involving the use of 4-methylumbelliferyl-β-D-glucuronide (MUG) as a fluorogenic substrate for enzyme reaction. Fluorescence intensity was negatively correlated (P < 0.001) with the volume of fresh citrus juice tested by the lauryl tryptose broth (LST)-MUG assay, and the permissible sample sizes were limited to 0.3 and 0.5 ml for fresh citrus juices with pHs of 3.3 and 3.9, respectively. In addition, false-negative results were visually observed under UV light when the E*Colite assay was used to test large volumes (5 to 10 ml per test) of fresh citrus juice or when the test broth used for the LSTMUG assay was supplemented with citric, malic, or tartaric acid at 2 to 4 g/liter. These results suggest that the size and pH of acidic samples should be controlled in MUG-based fluorogenic assays. The inhibitory effect on fluorescence was due to high acidity, which reduces fluorescence from 4-methylumbelliferone. Buffering improved the assays. When sodium bicarbonate was incorporated in the enrichment broth at 10 g/liter, the permissible sample sizes for fresh grapefruit juice (pH 3.1) increased from 0.3 to 1 ml for the LST-MUG (with 9.9 ml of broth) assay and from 3 to 10 ml for the E*Colite (with 99 ml of broth) assay.
Document Type: Research Article
Affiliations: 1: Virginia State University, Agricultural Research Station, P.O. Box 9061, Petersburg, Virginia 23806, USA 2: Florida Department of Citrus, Lake Alfred, Florida 33850, USA 3: University of Florida, Citrus Research and Education Center, Lake Alfred, Florida 33850, USA
Publication date: December 1, 2002
- IAFP Members with personal subscriptions to JFP Online: To access full-text JFP or JMFT articles, you must sign-in in the upper-right corner using your Ingenta sign-in details (your IAFP Member Login does not apply to this website). The Journal of Food Protection (JFP) is a refereed monthly publication. Each issue contains scientific research and authoritative review articles reporting on a variety of topics in food science pertaining to food safety and quality. The Journal is internationally recognized as the leading publication in the field of food microbiology with a readership exceeding 11,000 scientists from 70 countries. The Journal of Food Protection is indexed in Index Medicus, Current Contents, BIOSIS, PubMed, Medline, and many others.
Print and online subscriptions are available to IAFP Members and institutional subscribers. IAFP Members with a subscription to JFP Online will have access to all available JFP and JMFT content. Online visitors who are not IAFP Members or journal subscribers will be charged on a pay-per-view basis. Membership and subscription information is available at www.foodprotection.org.
- Information for Authors
- Submit a Paper
- Subscribe to this Title
- Membership Information
- Information for Advertisers
- Ingenta Connect is not responsible for the content or availability of external websites