Speciation of thermotolerant Campylobacter isolates involved in foodborne disease by means of DNA restriction analysis and molecular probes.
The molecular identification of several strains of Campylobacter jejuni and Campylobacter coli involved in foodborne disease was carried out by investigating the restriction profiles of their chromosomal DNA and by DNA/DNA hybridization. Cleavage with EcoRV allowed the visualization of a 3 kb DNA fragment characteristic of C. jejuni, whereas restriction with ClaI allowed the identification of a 9.3 kb DNA fragment, also characteristic of C. jejuni, and a DNA duplet of 9.5-10 kb, specific to C. coli. Restriction analysis with enzyme BglII allowed the visualization of DNA fragments of 3.5, 4, and 6.7 kb, characteristic of C. jejuni. C. jejuni subsp. doylei strains investigated shared a higher genetic homology among themselves-as determined by DNA/DNA hybridization-than with C. jejuni subsp. jejuni. A DNA probe, initially designed by Korolik et al. (Korolik, V.; Coloe, P. J.; Krishnapillai, V. J. Gen. Microbiol. 1988, 134, 521-529), including a DNA fragment encoding an antigenic membrane protein of 31.5 kDa in C. jejuni, when used as probe, allowed the specific identification of all strains of C. jejuni through the detection of strong hybridization signals in two BglII DNA fragments of 2.3 and 2.5 kb, which were not observed in C. coli. Cleavage of chromosomal DNA with BglII-either alone or coupled with probing assays with specific probes-proved to be a valuable tool for the speciation of Campylobacter isolates involved in foodborne disease.
Barros-Velázquez
Jorge
J
Departamento de Química Analítica, Nutrición y Bromatología, Facultad de Veterinaria, Campus Universitario Norte, Universidad de Santiago de Compostela, E-27002 Lugo, Spain.
Jiménez
Ana
A
Villa
Tomás G
TG
D002169:Campylobacter Infections / Q000382:microbiology; D017000:Campylobacter coli / Q000145:classification / Q000235:genetics*; D016123:Campylobacter jejuni / Q000145:classification / Q000235:genetics*; D015342:DNA Probes; D004262:DNA Restriction Enzymes; D004269:DNA, Bacterial / Q000235:genetics*; D005516:Food Microbiology; D005517:Foodborne Diseases / Q000382:microbiology; D005838:Genotype; D006801:Humans; D009693:Nucleic Acid Hybridization; D012150:Polymorphism, Restriction Fragment Length; D013045:Species Specificity
D015342:DNA Probes; D004269:DNA, Bacterial; D004262:DNA Restriction Enzymes