Agilent 2100 bioanalyzer for restriction fragment length polymorphism analysis of the Campylobacter jejuni flagellin gene.
The Agilent 2100 bioanalyzer (Agilent Technologies, Palo Alto, Calif.) utilizes capillary electrophoresis on a microchip device (LabChip 7500; Caliper Technologies, Mountain View, Calif.) that is capable of rapidly sizing small DNA fragments. To determine whether the system could replace conventional restriction fragment length polymorphism (RFLP) typing by agarose gel electrophoresis, we compared the analyzer with conventional flagellin RFLP for typing Campylobacter jejuni. Ninety-seven isolates representing 46 Fla types were initially analyzed. Correct Fla types were detected in 59% of the isolates. The major problem with the system was in resolving samples containing multiple DNA fragments differing from 8 to 20 bp. Overall, the bioanalyzer has the potential to replace conventional RFLP analysis by gel electrophoresis, but improvements in the chip separation are needed.
Nachamkin
I
I
Department of Pathology and Laboratory Medicine, University of Pennsylvania School of Medicine, Philadelphia, Pennsylvania 19104-4283, USA. nachamki@mail.med.upenn.edu
Panaro
N J
NJ
Li
M
M
Ung
H
H
Yuen
P K
PK
Kricka
L J
LJ
Wilding
P
P
D001322:Autoanalysis / Q000295:instrumentation / Q000379:methods; D016123:Campylobacter jejuni / Q000145:classification* / Q000235:genetics* / Q000302:isolation & purification; D019075:Electrophoresis, Capillary / Q000295:instrumentation / Q000379:methods; D004867:Equipment Design; D005408:Flagellin / Q000235:genetics*; D006801:Humans; D012150:Polymorphism, Restriction Fragment Length; D012703:Serotyping / Q000295:instrumentation / Q000379:methods
D005408:Flagellin