Computer identification of Escherichia coli rRNA gene restriction patterns.
A total of 191 strains of Escherichia coli comprising 164 serovar reference strains and 28 clinical strains were characterized by rRNA gene restriction patterns (ribotypes) generated after cleavage of total DNA with MluI, ClaI or HindIII restriction endonucleases and hybridization of fragments with acetylaminofluorene-labelled 16 + 23S rRNA. A wide diversity of ribotypes was observed with endonucleases MluI (104 patterns), ClaI (90 patterns) and HindIII (98 patterns). When MluI was used, 85% of patterns (11 to 15 fragments) shared five fragments 17.09, 3.94, 3.06, 2.23 and 1.76 kb in size. When these fragments were used as internal standards, the percent errors in fragment length determination was half of that obtained with an external standard. Two fragment size databases of MluI and ClaI ribotypes were built. Automatic identification was obtained after setting the percent fragment size variation tolerance (error) at 5%. MluI ribotyping is recommended as a primary epidemiological marker. Strains with similar MluI ribotype should then be submitted to ClaI ribotyping. Ribotyping with HindIII can only be the third choice, since the patterns were often uncertain due to the frequent occurrence of faint bands. Most of the studied serovars gave discrete patterns and these data provide the basis for a molecular typing system for E. coli which could possibly substitute for serotyping when the latter is not available.
Machado
J
J
Centre National de Typage Moléculaire des Entérobactéries, Unité des Entérobactéries, INSERM U389, Paris.
Grimont
F
F
Grimont
P A
PA
D015073:2-Acetylaminofluorene / Q000737:chemistry; D001426:Bacterial Proteins; D015373:Bacterial Typing Techniques; D004262:DNA Restriction Enzymes / Q000378:metabolism*; D004269:DNA, Bacterial / Q000737:chemistry*; D015247:Deoxyribonuclease HindIII / Q000378:metabolism; D015252:Deoxyribonucleases, Type II Site-Specific / Q000378:metabolism; D001330:Electronic Data Processing / Q000379:methods*; D004587:Electrophoresis, Agar Gel; D004926:Escherichia coli / Q000737:chemistry / Q000145:classification* / Q000235:genetics; D020459:Genes, rRNA / Q000235:genetics*; D006801:Humans; D007124:Immunoenzyme Techniques; D012150:Polymorphism, Restriction Fragment Length; D012336:RNA, Ribosomal, 16S / Q000737:chemistry / Q000235:genetics; D012338:RNA, Ribosomal, 23S / Q000737:chemistry / Q000235:genetics; D012680:Sensitivity and Specificity; D012703:Serotyping
D001426:Bacterial Proteins; D004269:DNA, Bacterial; D012336:RNA, Ribosomal, 16S; D012338:RNA, Ribosomal, 23S; D015073:2-Acetylaminofluorene; D004262:DNA Restriction Enzymes; D015247:Deoxyribonuclease HindIII; C035041:endodeoxyribonuclease MluI; C032824:ATCGAT-specific type II deoxyribonucleases; D015252:Deoxyribonucleases, Type II Site-Specific