Identification and classification of Oxalobacter formigenes strains by using oligonucleotide probes and primers.
Genomic DNAs of various strains of Oxalobacter formigenes were subjected to restriction endonuclease fragment length polymorphism- and PCR-based amplification analyses with DNA probes and primers complementary to sequences within either the oxc gene, encoding oxalyl coenzyme A (oxalyl-CoA) decarboxylase, or the frc gene, encoding formyl-CoA transferase. Oligonucleotide probes based on nonconserved sequences of oxc or frc were able to divide O. formigenes strains into at least two groups, consistent with the current separation of O. formigenes strains into groups I and II on the basis of 16S rRNA sequence similarities and lipid content. In contrast, an oligonucleotide probe based on the conserved 5' end of oxc appeared to bind all group I and the majority of group II strains. PCR amplification of the oxc gene showed even greater sensitivity in detecting O. formigenes and provided support for further division of the strains into subgroups. In addition, these oligonucleotides failed to hybridize to or amplify PCR products from whole fecal DNA isolated from fresh stool samples from an individual not colonized with O. formigenes, indicating unique specificity. Thus, these DNA analyses permit both detection as well as classification of O. formigenes strains.
Sidhu
H
H
Department of Pathology & Laboratory Medicine, University of Florida, Gainesville 32610, USA.
Allison
M
M
Peck
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AB
D015373:Bacterial Typing Techniques; D002262:Carboxy-Lyases / Q000235:genetics; D015256:Coenzyme A-Transferases / Q000235:genetics; D017124:Conserved Sequence; D017931:DNA Primers; D004269:DNA, Bacterial / Q000032:analysis / Q000235:genetics; D005798:Genes, Bacterial; D006089:Gram-Negative Anaerobic Bacteria / Q000145:classification* / Q000235:genetics / Q000302:isolation & purification*; D015345:Oligonucleotide Probes; D016133:Polymerase Chain Reaction; D012150:Polymorphism, Restriction Fragment Length; D012680:Sensitivity and Specificity
D017931:DNA Primers; D004269:DNA, Bacterial; D015345:Oligonucleotide Probes; D015256:Coenzyme A-Transferases; C064813:formyl-coenzyme A transferase; D002262:Carboxy-Lyases; C059418:oxalyl CoA decarboxylase