Report on overcoming the poor quality of Apai pulsotypes with a short review on PFGE for listeria monocytogenes.
Since listeriosis, caused by Listeria monocytogenes, is one of the important concerns of public health in Europe related to foodborne zoonoses, an efficient protocol for isolate typing is necessary when performing epidemiological studies. Three standardized PFGE protocols available for L. monocytogenes were briefly reviewed. Since observing a poor-quality of ApaI pulsotypes in our laboratory, enzymes from three different manufacturers were compared. The obtained pulsotypes showed that restriction digestion with ApaI from New England BioLabs should be complemented with a subsequent overnight incubation of PFGE plugs in TE buffer for better performance.
Institute of Microbiology and Parasitology, Veterinary Faculty, University of Ljubljana, Ljubljana, Slovenia. darja.kusar@vf.uni-lj.si
Darja
D
Kusar
Institute of Microbiology and Parasitology, Veterinary Faculty, University of Ljubljana, Ljubljana, Slovenia.
Maja
M
Kavalic
Institute of Microbiology and Parasitology, Veterinary Faculty, University of Ljubljana, Ljubljana, Slovenia.
Matjaz
M
Ocepek
Institute of Microbiology and Parasitology, Veterinary Faculty, University of Ljubljana, Ljubljana, Slovenia.
Irena
I
Zdovc
D004269:DNA, Bacterial / Q000235:genetics; D015252:Deoxyribonucleases, Type II Site-Specific / Q000737:chemistry*; D016521:Electrophoresis, Gel, Pulsed-Field / Q000295:instrumentation / Q000379:methods*; D006801:Humans; D008089:Listeria monocytogenes / Q000145:classification / Q000235:genetics / Q000302:isolation & purification*; D008088:Listeriosis / Q000382:microbiology; D011933:Reagent Kits, Diagnostic; D012703:Serotyping / Q000295:instrumentation / Q000379:methods*
D004269:DNA, Bacterial; D011933:Reagent Kits, Diagnostic; D015252:Deoxyribonucleases, Type II Site-Specific; C038485:GGGCCC-specific type II deoxyribonucleases