Analysis of PCR-based methods for characterization of Listeria monocytogenes strains isolated from different sources.
Listeria monocytogenes strains, isolated from various sources (food, environment, and animals), were used to test different PCR-based methods to investigate their capability to define the strain origin. RAPD-PCR with three primers and the SAU-PCR method, in which the DNA was first digested with the Sau3A restriction endonuclease and then amplified with a primer designed on the restriction site, were carried out, and the profiles obtained were used to perform cluster analysis. Based on the cluster analysis of Listeria spp. strains, obtained from international collections, the coefficient of similarity was selected. The results obtained showed that the methods tested in the study gave different levels of differentiation between the strains tested. The RAPD protocol using the P1254 primer and the SAU-PCR gave appreciable results only for strains isolated from animals and from a food processing plant in two different periods of the year 2003. Better differentiation was observed using the RAPD-PCR with primer D8635. As a matter of fact, it was able to distinguish L. monocytogenes obtained from different species of animals, different food samples and strains from the same production plant isolated in different periods of the year. Also primer M13 gave positive results, but the coefficient of similarity to use had to be increased to 80%. On the basis of the results observed, RAPD-PCR with primers D8635 and M13 should be considered reliable tools for epidemiological investigations focusing on L. monocytogenes.
Cocolin
Luca
L
Dipartimento di Scienze degli Alimenti, Università degli studi di Udine, via Marangoni 97, 33100, Udine, Italy. lscocolin@uniud.it
Stella
Simone
S
Nappi
Raffaella
R
Bozzetta
Elena
E
Cantoni
Carlo
C
Comi
Giuseppe
G
D016000:Cluster Analysis; D017931:DNA Primers; D004269:DNA, Bacterial / Q000032:analysis*; D005506:Food Contamination / Q000032:analysis; D005516:Food Microbiology; D008089:Listeria monocytogenes / Q000145:classification* / Q000302:isolation & purification*; D010802:Phylogeny; D019105:Random Amplified Polymorphic DNA Technique / Q000379:methods*; D012680:Sensitivity and Specificity; D013045:Species Specificity
D017931:DNA Primers; D004269:DNA, Bacterial