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Design of a New Universal Real-Time PCR System Targeting the tuf Gene for the Enumeration of Bacterial Counts in Food
- Source :
- Journal of Food Protection. 73:670-679
- Publication Year :
- 2010
- Publisher :
- Elsevier BV, 2010.
-
Abstract
- A novel universal real-time PCR, consisting of newly designed oligonucleotide subsets, was designed for a bacterial housekeeping gene encoding the peptide elongation factor Tu. Specificity and universality were confirmed in 66 bacterial strains, including 51 genera and 63 species. The amplification kinetics of tuf gene–targeted real-time quantitative PCR were consistent in a wide range of bacterial species tested. A calibration curve (r2 = 0.97) was produced for the estimation of bacterial counts, based on measurements of representative inoculations with 10-fold serial dilutions of the cells of representative bacterial species. Linear regression analysis of the real-time PCR–derived bacterial counts and aerobic plate counts, in a total 149 samples consisting of 25 minced meat, 34 fresh-cut vegetables, and 90 fish, exhibited a high correlation (r2 = 0.84, 0.87, and 0.95, respectively) over the range of 3.0 to 9.0 log CFU/g. In total, the difference between the two methods was less than 0.5 log in 75 of these samples, and in the remaining 74 samples, the difference was 0.5 to 1.0 log. Presently, our tuf gene–targeted real-time quantitative PCR assay achieves a rapid (within 2 h) estimation of bacterial counts of 3.0 to 9.0 log CFU/g, in a practical manner.
- Subjects :
- DNA, Bacterial
Time Factors
Serial dilution
Colony Count, Microbial
Food Contamination
Peptide Elongation Factor Tu
Biology
Polymerase Chain Reaction
Sensitivity and Specificity
Microbiology
Bacterial counts
law.invention
Species Specificity
law
Enumeration
Humans
Food microbiology
Polymerase chain reaction
Bacteria
Tuf gene
Gene Amplification
Housekeeping gene
Real-time polymerase chain reaction
Consumer Product Safety
Food Microbiology
Linear Models
Food Science
Subjects
Details
- ISSN :
- 19449097 and 0362028X
- Volume :
- 73
- Database :
- OpenAIRE
- Journal :
- Journal of Food Protection
- Accession number :
- edsair.doi.dedup.....fa070dc2c5a2d21972fcfb1e2b1e60e2