Rapid identification and classification of bacteria by 16S rDNA restriction fragment melting curve analyses (RFMCA).
The aim of this work was to evaluate restriction fragment melting curve analyses (RFMCA) as a novel approach for rapid classification of bacteria during food production. RFMCA was evaluated for bacteria isolated from sous vide food products, and raw materials used for sous vide production. We identified four major bacterial groups in the material analysed (cluster I-Streptococcus, cluster II-Carnobacterium/Bacillus, cluster III-Staphylococcus and cluster IV-Actinomycetales). The accuracy of RFMCA was evaluated by comparison with 16S rDNA sequencing. The strains satisfying the RFMCA quality filtering criteria (73%, n=57), with both 16S rDNA sequence information and RFMCA data (n=45) gave identical group assignments with the two methods. RFMCA enabled rapid and accurate classification of bacteria that is database compatible. Potential application of RFMCA in the food or pharmaceutical industry will include development of classification models for the bacteria expected in a given product, and then to build an RFMCA database as a part of the product quality control.
Rudi
Knut
K
MATFORSK Norwegian Food Research Institute, Oslo, Norway. knut.rudi@matforsk.no
Kleiberg
Gro H
GH
Heiberg
Ragnhild
R
Rosnes
Jan T
JT
D001419:Bacteria / Q000145:classification / Q000235:genetics / Q000302:isolation & purification; D001483:Base Sequence; D004269:DNA, Bacterial / Q000032:analysis / Q000737:chemistry / Q000235:genetics*; D005506:Food Contamination / Q000032:analysis*; D005516:Food Microbiology; D010802:Phylogeny; D012336:RNA, Ribosomal, 16S / Q000032:analysis / Q000737:chemistry / Q000235:genetics*; D015203:Reproducibility of Results; D015183:Restriction Mapping / Q000379:methods*; D012680:Sensitivity and Specificity; D017422:Sequence Analysis, DNA / Q000379:methods*; D044366:Transition Temperature
D004269:DNA, Bacterial; D012336:RNA, Ribosomal, 16S