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Title: RAPID ESTIMATION OF SPOILAGE BACTERIAL LOAD IN AEROBICALLY STORED MEAT BY A QUANTITATIVE POLYMERASE CHAIN REACTION

Author
item VENKITANARAYANAN, KUMAR - UNIV OF CONNECTICUT
item FAUSTMAN, CAMERON - UNIV OF CONNECTICUT
item CRIVELLO, JOSEPH - UNIV OF CONNECTICUT
item KHAN, MOHAMMED - UNIV OF CONNECTICUT
item HOAGLAND, THOMAS - UNIV OF CONNECTICUT
item Berry, Bradford

Submitted to: Journal of Applied Microbiology
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 9/20/1997
Publication Date: N/A
Citation: N/A

Interpretive Summary: Two major groups of bacteria, pathogenic and spoilage, are responsible for significant food safety concern and monetary loss in the meat industry. Rapid and accurate testing procedures are needed for identifying these two groups of microorganisms before meat products are purchased by consumers. Previously, DNA probes have been successfully used in rapidly detecting food-borne pathogens. However, little research has been performed to determine the usefulness of this approach for spoilage bacteria. Based on 23S DNA of Pseudomas aeruginosa, two primers were synthesized for a Polymerase Chain Reaction. Using nine different meat spoilage bacteria, a unique 207 basepair DNA product was identified. Using a DNA probe specific for this product, it was confirmed that the DNA sequence was unique for the spoilage bacteria studied. A quantification of the polymerase chain reaction product was conducted by electrochemiluminescence. Values for the quantification product were highly related to aerobic plate counts. Since this test procedure requires only four hours to complete and was very sensitive with high bacterial numbers, it should prove to be very effective in HAACP programs.

Technical Abstract: We report a Quantitative PCR which utilizes primers from a conserved 23S rDNA sequence identified in nine different spoilage bacteria commonly present in meat. The PCR detected the spoilage bacteria by amplifying a specific 207 bp sequence from their chromosomal DNA. Quantification of PCR product by electrochemiluminescence revealed that the concentration of the amplified product was dependent on cycle number and the initial number of bacteria present in the sample. Statistical analysis of the results indicated a correlation of coefficient of 0.94 (P<0.001) between aerobic plate count and OPCR luminosity units.