Use of an amplified-fragment length polymorphism technique to fingerprint and differentiate isolates of Helicobacter pylori.
Amplified-fragment length polymorphism (AFLP) analysis is the name given to a genotypic technique in which adapter oligonucleotides are ligated to restriction enzyme fragments and then used as target sites for primers in a PCR amplification process. The amplified fragments are electrophoretically separated to give strain-specific band profiles. We have developed a single-enzyme approach that did not require costly equipment or reagents for the fingerprinting of strains of Helicobacter pylori. The method was assessed with 46 isolates of H. pylori from 28 patients, and the results were compared with those from other genotypic tests. The AFLP profiles derived from HindIII fragments differentiated strains of H. pylori from unrelated individuals and confirmed the common origin of strains in some family members. AFLP analysis was also applied to investigate persistent infection following antibiotic therapy. Overall, the modified technique was relatively rapid and technically simple yet gave reproducible and discriminatory results. AFLP analysis samples variation throughout the genome and is a valuable addition to the existing genotypic fingerprinting methods for H. pylori.
Helicobacter Reference Unit, Laboratory of Enteric Pathogens, Central Public Health Laboratory, London NW9 5HT, United Kingdom.
J R
JR
Gibson
E
E
Slater
J
J
Xerry
D S
DS
Tompkins
R J
RJ
Owen
D000293:Adolescent; D000328:Adult; D001483:Base Sequence; D002648:Child; D017931:DNA Primers; D004269:DNA, Bacterial / Q000737:chemistry / Q000235:genetics; D015247:Deoxyribonuclease HindIII; D005260:Female; D005753:Gastric Mucosa / Q000382:microbiology; D005838:Genotype; D016481:Helicobacter Infections / Q000382:microbiology / Q000473:pathology; D016480:Helicobacter pylori / Q000145:classification* / Q000235:genetics* / Q000302:isolation & purification; D006801:Humans; D008297:Male; D008826:Microbial Sensitivity Tests; D009679:Nuclear Family; D016133:Polymerase Chain Reaction / Q000379:methods; D012150:Polymorphism, Restriction Fragment Length; D015183:Restriction Mapping; D013698:Templates, Genetic
D017931:DNA Primers; D004269:DNA, Bacterial; D015247:Deoxyribonuclease HindIII