Differentiation of Chlamydia psittaci and C. pecorum strains by species-specific PCR.
Sequence analyses 5' ends of the 60-kDa cysteine-rich outer membrane protein genes (Omp2) of Chlamydia psittaci and Chlamydia pecorum strains indicate that these species have approximately 70% nucleotide identity. On the basis of this sequence information, PCR primers were designed to allow the specific amplification of DNA extracted from C. psittaci S26/3 (abortion strain), P94/1 (pigeon strain), and C. pecorum W73 (fecal strain) in one reaction tube. By using nested reactions (with primers PCR-D1 and PCR-D2 followed by the specific primers and PCR-D2), 0.6, 0.2, and 8 inclusion-forming units of S26/3, P94/1 (both diluted in tissue culture-negative placental material), and W73 (diluted in culture-negative fecal material) per ml, respectively, were detected. The differentiation of C. psittaci and C. pecorum strains of ovine and bovine origins was carried out, and the results were in agreement with those obtained from AluI restriction enzyme analysis of DNA amplified from corresponding strains by PCR. This approach allows the simultaneous detection and typing of C. psittaci and C. pecorum strains and the identification of samples containing both species.
Sheehy
N
N
Department of Veterinary Microbiology and Parasitology, Faculty of Veterinary Medicine, University College Dublin, Ireland.
Markey
B
B
Gleeson
M
M
Quinn
P J
PJ
D000595:Amino Acid Sequence; D000818:Animals; D001425:Bacterial Outer Membrane Proteins / Q000235:genetics; D015373:Bacterial Typing Techniques; D001431:Bacteriological Techniques; D001483:Base Sequence; D002417:Cattle; D002642:Chick Embryo; D002689:Chlamydia / Q000145:classification* / Q000235:genetics* / Q000302:isolation & purification; D002691:Chlamydophila psittaci / Q000235:genetics* / Q000302:isolation & purification / Q000472:pathogenicity; D004269:DNA, Bacterial / Q000235:genetics; D005069:Evaluation Studies as Topic; D005798:Genes, Bacterial; D006801:Humans; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction / Q000379:methods*; D017386:Sequence Homology, Amino Acid; D012689:Sequence Homology, Nucleic Acid; D013045:Species Specificity
D001425:Bacterial Outer Membrane Proteins; D004269:DNA, Bacterial; C063931:Omp2 protein, bacteria