Differentiation of closely related Carnobacterium food isolates based on 16S-23S ribosomal DNA intergenic spacer region polymorphism.
A novel strategy for identification of Carnobacterium food isolates based on restriction fragment length polymorphism (RFLP) of PCR-amplified 16S-23S ribosomal intergenic spacer regions (ISRs) was developed. PCR amplification from all Carnobacterium strains studied always yielded three ISR amplicons, which were designated the small ISR (S-ISR), the medium ISR (M-ISR), and the large ISR (L-ISR). The lengths of these ISRs varied from one species to another. Carnobacterium divergens NCDO 2763(T) and C. mobile DSM 4849(T) generated one major S-ISR band (ca. 400 bp) and minor M-ISR and L-ISR bands (ca. 500 and ca. 600 bp, respectively). The ISRs amplified from C. gallinarum NCFB 2766(T) and C. piscicola NCDO 2762(T) were larger (S-ISR, ca. 600 bp; M-ISR, ca. 700 bp; and L-ISR, ca. 800 bp). The L-ISR contained two tDNAs coding for tRNA(Ile) and tRNA(Ala) genes. The M-ISR included one tRNA(Ala) gene, and the S-ISR did not contain a tDNA gene. The RFLP scheme devised involves estimation of variable PCR product sizes together with HinfI, TaqI, and HindIII restriction analysis. Forty-two isolates yielded four unique band patterns that correctly resolved these isolates into four Carnobacterium species. This method is very suitable for rapid, low-cost identification of a wide variety of Carnobacterium species without sequencing.
Kabadjova
Petia
P
Laboratoire de Microbiologie Alimentaire et Industrielle, ENITIAA, F-44322 Nantes Cedex 3, France.
Dousset
Xavier
X
Le Cam
Virginie
V
Prevost
Hervé
H
D001419:Bacteria / Q000145:classification / Q000235:genetics / Q000302:isolation & purification*; D001483:Base Sequence; D004269:DNA, Bacterial / Q000032:analysis*; D021901:DNA, Intergenic / Q000032:analysis / Q000235:genetics*; D008969:Molecular Sequence Data; D011110:Polymorphism, Genetic; D012335:RNA, Ribosomal / Q000032:analysis / Q000235:genetics*; D012689:Sequence Homology, Nucleic Acid
D004269:DNA, Bacterial; D021901:DNA, Intergenic; D012335:RNA, Ribosomal