TY - JOUR
T1 - Repetitive element sequence-based pcr typing for improved discrimination of Campylobacter Jejuni
AU - Malakauskas, Mindaugas
AU - Malakauskas, Alvydas
AU - Christensen, Henrik
AU - Olsen, John Elmerdahl
AU - Brogren, Carl Henrik
PY - 2017
Y1 - 2017
N2 - The aim of our studies was to evaluate rep-PCR typing using different primers as a fast genotyping method for discrimination of C. jejuni. Also a microfluidic electrophoretic DNA separation method for rapid DNA fingerprinting based on Lab-on-a-Chip technology was tested. Based on the test results, the single polytrinucleotide (GTG)5 was chosen as primer. Nine strains representing three epidemiological groups from the CAMPYNET network strains were assigned to three different groups by both (GTG)5- PCR and PFGE. Typing of 72 epidemiologically independent strains using (GTG)5-PCR and PFGE clustered the strains in 60 and 53 clusters, respectively, with an identical discriminatory power (D=0,99). When PCR-products were separated on the Agilent 2100 Bioanalyzer Lab-on-a-Chip device the strains were grouped with the same discriminatory power, though some strains were clustered differently compare to gel-based DNA amplicon separation. The main disadvantage of (GTG)5-PCR typing was differences in band intensity that complicates comparison of (GTG)5-PCR fingerprints when numerous strains are evaluated. The advantages of rep-PCR based subtyping were rapidity, simplicity, similar discriminatory power to PFGE and possible rapid DNA amplicon separation based on Lab-on-a-Chip technology with Agilent 2100 Bioanalyzer.
AB - The aim of our studies was to evaluate rep-PCR typing using different primers as a fast genotyping method for discrimination of C. jejuni. Also a microfluidic electrophoretic DNA separation method for rapid DNA fingerprinting based on Lab-on-a-Chip technology was tested. Based on the test results, the single polytrinucleotide (GTG)5 was chosen as primer. Nine strains representing three epidemiological groups from the CAMPYNET network strains were assigned to three different groups by both (GTG)5- PCR and PFGE. Typing of 72 epidemiologically independent strains using (GTG)5-PCR and PFGE clustered the strains in 60 and 53 clusters, respectively, with an identical discriminatory power (D=0,99). When PCR-products were separated on the Agilent 2100 Bioanalyzer Lab-on-a-Chip device the strains were grouped with the same discriminatory power, though some strains were clustered differently compare to gel-based DNA amplicon separation. The main disadvantage of (GTG)5-PCR typing was differences in band intensity that complicates comparison of (GTG)5-PCR fingerprints when numerous strains are evaluated. The advantages of rep-PCR based subtyping were rapidity, simplicity, similar discriminatory power to PFGE and possible rapid DNA amplicon separation based on Lab-on-a-Chip technology with Agilent 2100 Bioanalyzer.
KW - Campylobacter
KW - Genotyping
KW - Rep-PCR
M3 - Journal article
AN - SCOPUS:85030530790
SN - 1392-2130
VL - 75
SP - 43
EP - 53
JO - Veterinarija ir Zootechnika
JF - Veterinarija ir Zootechnika
IS - 97
ER -