Evaluation of ribotyping techniques as applied to Arcobacter, Campylobacter and Helicobacter.
Restriction fragment length polymorphisms of ribosomal DNA (ribotyping) of many bacterial species has been useful for both epidemiologic subtyping and species identification. However, the use of ribotyping has been confined to major research and reference laboratories due to two factors: (a) the procedure must be carefully optimized for each organism one wishes to investigate and (b) most currently available protocols use hazardous chemicals or radioisotopes. The purpose of this study is to suggest an overall scheme that a clinical or research microbiologist could apply to ribotyping of any organism. In general, we recommend using a guanidium extraction method for DNA extraction, careful optimization of restriction conditions, and hybridization with non-radioactive digoxigenin-labelled probes; these procedures do not use hazardous chemicals or radioisotopes.
Kiehlbauch
J A
JA
Enteric Diseases Branch, Centers for Disease Control and Prevention, Atlanta, GA 30333.
Cameron
D N
DN
Wachsmuth
I K
IK
D015139:Blotting, Southern; D002167:Campylobacter / Q000145:classification / Q000235:genetics*; D004269:DNA, Bacterial / Q000032:analysis / Q000235:genetics*; D004275:DNA, Ribosomal / Q000032:analysis / Q000235:genetics*; D004076:Digoxigenin; D016998:Helicobacter / Q000145:classification / Q000235:genetics*; D008828:Microbiological Techniques; D012150:Polymorphism, Restriction Fragment Length
D004269:DNA, Bacterial; D004275:DNA, Ribosomal; D004076:Digoxigenin