RRC ID 29462
著者 Ramachandran P, Lacher DW, Pfeiler EA, Elkins CA.
タイトル Development of a tiered multilocus sequence typing scheme for members of the Lactobacillus acidophilus complex.
ジャーナル Appl Environ Microbiol
Abstract Members of the Lactobacillus acidophilus complex are associated with functional foods and dietary supplements because of purported health benefits they impart to the consumer. Many characteristics of these microorganisms are reported to be strain specific. Therefore, proper strain typing is essential for safety assessment and product labeling, and also for monitoring strain integrity for industrial production purposes. Fifty-two strains of the L. acidophilus complex (L. acidophilus, L. amylovorus, L. crispatus, L. gallinarum, L. gasseri, and L. johnsonii) were genotyped using two established methods and compared to a novel multilocus sequence typing (MLST) scheme. PCR restriction fragment length polymorphism (PCR-RFLP) analysis of the hsp60 gene with AluI and TaqI successfully clustered 51 of the 52 strains into the six species examined, but it lacked strain-level discrimination. Random amplified polymorphic DNA PCR (RAPD-PCR) targeting the M13 sequence resulted in highly discriminatory profiles but lacked reproducibility. In this study, an MLST scheme was developed using the conserved housekeeping genes fusA, gpmA, gyrA, gyrB, lepA, pyrG, and recA, which identified 40 sequence types that successfully clustered all of the strains into the six species. Analysis of the observed alleles suggests that nucleotide substitutions within five of the seven MLST loci have reached saturation, a finding that emphasizes the highly diverse nature of the L. acidophilus complex and our unconventional application of a typically intraspecies molecular typing tool. Our MLST results indicate that this method could be useful for characterization and strain discrimination of a multispecies complex, with the potential for taxonomic expansion to a broader collection of Lactobacillus species.
巻・号 79(23)
ページ 7220-8
公開日 2013-12-1
DOI 10.1128/AEM.02257-13
PII AEM.02257-13
PMID 24038697
PMC PMC3837765
MeSH Cluster Analysis DNA, Bacterial / chemistry DNA, Bacterial / genetics Food Microbiology / methods* Genes, Essential Genotype Lactobacillus acidophilus / classification* Lactobacillus acidophilus / genetics* Molecular Sequence Data Multilocus Sequence Typing / methods* Sequence Analysis, DNA
IF 4.016
引用数 10
WOS 分野 BIOTECHNOLOGY & APPLIED MICROBIOLOGY MICROBIOLOGY
リソース情報
一般微生物 JCM 5810