PCR detection and molecular identification of Chlamydiaceae species.
Recent taxonomic developments, based on 16s and 23s rRNA gene sequences, have divided the family Chlamydiaceae into two genera and nine species, of which five have been found to infect humans. Few simple methods are available to detect and identify all species sensitively and specifically. In this study the suitability of the omp2 gene as a target for molecular identification of Chlamydiaceae is demonstrated. Phylogenetic analysis of partial omp2 gene sequences from all nine species agrees with the recently published taxonomic changes based on the ribosomal genes. The use of a family-specific PCR primer pair, which is able to amplify the 5' end of the omp2 gene from all Chlamydiaceae except some Chlamydophila pecorum strains, is described. Identification of all nine species was achieved using restriction fragment length polymorphism analysis with a single enzyme, AluI, confirmed by DNA sequencing. A PCR enzyme-linked oligonucleotide assay was developed which can detect a single chlamydial genome and may be applied to DNA extracts from any specimen or culture for the detection of single or mixed human chlamydial infection.
Hartley
J C
JC
Department of Clinical Microbiology, University College London Hospital NHS Trust, London WC1E 6DB, United Kingdom.
Kaye
S
S
Stevenson
S
S
Bennett
J
J
Ridgway
G
G
D000818:Animals; D001425:Bacterial Outer Membrane Proteins / Q000235:genetics*; D001483:Base Sequence; D002693:Chlamydiaceae / Q000145:classification* / Q000235:genetics / Q000302:isolation & purification*; D016000:Cluster Analysis; D017931:DNA Primers / Q000235:genetics; D015252:Deoxyribonucleases, Type II Site-Specific; D006801:Humans; D008969:Molecular Sequence Data; D010802:Phylogeny; D016133:Polymerase Chain Reaction / Q000379:methods*; D012150:Polymorphism, Restriction Fragment Length; D012680:Sensitivity and Specificity; D017422:Sequence Analysis, DNA; D013045:Species Specificity
D001425:Bacterial Outer Membrane Proteins; D017931:DNA Primers; C063931:Omp2 protein, bacteria; C024669:endodeoxyribonuclease AluI; D015252:Deoxyribonucleases, Type II Site-Specific