Comparative fingerprinting analysis of Campylobacter jejuni subsp. jejuni strains by amplified-fragment length polymorphism genotyping.
Amplified-fragment length polymorphism (AFLP) analysis with the endonucleases BglII and MfeI was used to genotype 91 Campylobacter jejuni subsp. jejuni strains from outbreaks and sporadic cases. AFLP-generated fragments were labeled with fluorescent dye and separated by capillary electrophoresis. The software packages GeneScan and GelCompar II were used to calculate AFLP pattern similarities and to investigate phylogenetic relationships among the genotyped strains. The AFLP method was compared with two additional DNA-based typing methods, pulsed-field gel electrophoresis (PFGE) using SmaI and restriction fragment length polymorphism analysis on PCR products (PCR-RFLP) of the flaA and flaB genes. We found that AFLP analysis of C. jejuni strains is a rapid method that offers better discriminatory power than do both PFGE and PCR-RFLP. AFLP and, to a lesser extent, PCR-RFLP could differentiate strains within the same PFGE profiles, which also makes PCR-RFLP an alternative to PFGE. We were able to clearly distinguish 9 of 10 recognized outbreaks by AFLP and to identify similarities among outbreak and sporadic strains. Therefore, AFLP is suitable for epidemiological surveillance of C. jejuni and will be an excellent tool for source identification in outbreak situations.
Lindstedt
B A
BA
National Institute of Public Health, Department of Bacteriology, N-0403 Oslo, Norway. bjorn-arne.lindstedt@folkehelsa.no
Heir
E
E
Vardund
T
T
Melby
K K
KK
Kapperud
G
G
D000818:Animals; D001426:Bacterial Proteins; D002169:Campylobacter Infections / Q000382:microbiology; D016123:Campylobacter jejuni / Q000145:classification* / Q000235:genetics*; D016172:DNA Fingerprinting / Q000379:methods*; D004269:DNA, Bacterial / Q000032:analysis; D016208:Databases, Factual; D015252:Deoxyribonucleases, Type II Site-Specific / Q000378:metabolism; D016521:Electrophoresis, Gel, Pulsed-Field; D005838:Genotype; D006801:Humans; D016133:Polymerase Chain Reaction; D012150:Polymorphism, Restriction Fragment Length; D015183:Restriction Mapping
D001426:Bacterial Proteins; D004269:DNA, Bacterial; C059776:endodeoxyribonuclease MfeI; C024404:BglII endonuclease; D015252:Deoxyribonucleases, Type II Site-Specific