Abstract：
:An oligonucleotide array based on PCR method containing a combination of probes for taxonomic markers and species-specific virulence genes was developed for the simultaneous identification of 14 Lactic Acid Bacteria (LAB) and food-borne pathogenic bacteria in raw milk. The hypervariable regions V3 and V6 of 16S, together with a variable region of 23S rRNA genes and several genes specific for virulence factors were selected. Universal primers and multiplex PCR were used for the rapid differentiation of bacterial species and low concentration of specific pathogenic and spoilage bacteria were detected in milk. The dominant species such as Streptococcus thermophilus, Enterococcus faecalis, Lactococcus lactis, andLeuconostoc lactis were identified by indirect-labelling reactions based upon incorporation of amino-allyl dUTPs. The results regarding food-borne pathogens detection showed highly specific hybridisation patterns with the genomic DNA from Campylobacter jejuni, Escherichia coli, Salmonella thyphimurium, Listeria monocytogenes and Yersinia enterocolitica. A clear differentiation of dominant species present in a complex microbial community such as raw milk was achieved by the application of short oligonucleotide probes which discriminate sequences differing by few nucleotides.
journal_name
J Microbiol Methodsjournal_title
Journal of microbiological methodsauthors
Giannino ML,Aliprandi M,Feligini M,Vanoni L,Brasca M,Fracchetti Fdoi
10.1016/j.mimet.2009.05.015subject
Has Abstractpub_date
2009-08-01 00:00:00pages
181-8issue
2eissn
0167-7012issn
1872-8359pii
S0167-7012(09)00154-7journal_volume
78pub_type
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