R D Wagner1, D D Paine, C E Cerniglia. 1. Microbiology Division, FDA National Center for Toxicological Research, Jefferson, AR 72079, USA. dagner@nctr.fda.gov
Abstract
AIMS: Phenotypic and genotypic bacteria identification methods were compared for their efficacy in determining the composition of competitive exclusion (CE) products. METHODS AND RESULTS: Phenotypic methods used for bacterial identification were fatty acid methyl ester profiles, biochemical assays and carbohydrate utilization profiles. Genotypic methods were MicroSeq16S rRNA sequence analysis and BLAST searches of the GenBank sequence database. Agreement between phenotypic and genotypic methods for identification of bacteria isolated from the Preempt CE product was 20%. A defined test mixture of bacteria was identified to the species level 100% by BLAST analysis, 64% by MicroSeq and 36% by phenotypic techniques. CONCLUSIONS: The wide range of facultative and obligate anaerobic bacteria present in a CE product are more accurately identified with 16S rRNA sequence analyses than with phenotypic identification techniques. SIGNIFICANCE AND IMPACT OF THE STUDY: These results will provide guidelines for manufacturers of CE products to submit more reliable product information for market approval by regulatory agencies.
AIMS: Phenotypic and genotypic bacteria identification methods were compared for their efficacy in determining the composition of competitive exclusion (CE) products. METHODS AND RESULTS: Phenotypic methods used for bacterial identification were fatty acid methyl ester profiles, biochemical assays and carbohydrate utilization profiles. Genotypic methods were MicroSeq16S rRNA sequence analysis and BLAST searches of the GenBank sequence database. Agreement between phenotypic and genotypic methods for identification of bacteria isolated from the Preempt CE product was 20%. A defined test mixture of bacteria was identified to the species level 100% by BLAST analysis, 64% by MicroSeq and 36% by phenotypic techniques. CONCLUSIONS: The wide range of facultative and obligate anaerobic bacteria present in a CE product are more accurately identified with 16S rRNA sequence analyses than with phenotypic identification techniques. SIGNIFICANCE AND IMPACT OF THE STUDY: These results will provide guidelines for manufacturers of CE products to submit more reliable product information for market approval by regulatory agencies.
Authors: Anita Menconi; Gopala Kallapura; Juan D Latorre; Marion J Morgan; Neil R Pumford; Billy M Hargis; Guillermo Tellez Journal: Biosci Microbiota Food Health Date: 2014-01-30
Authors: Javier Fernando Melo-Bolívar; Ruth Yolanda Ruiz Pardo; Michael E Hume; David J Nisbet; Fernando Rodríguez-Villamizar; Juan F Alzate; Howard Junca; Luisa Marcela Villamil Díaz Journal: PLoS One Date: 2019-05-03 Impact factor: 3.240