PCR-based detection of chlamydial infection in swine and subsequent PCR-coupled genotyping of chlamydial omp1-gene amplicons by DNA-hybridization, RFLP-analysis, and nucleotide sequence analysis.
Lung and intestine of 49 pigs with respiratory diseases and endocervical swabs from 205 sows with reproductive disorders were investigated for chlamydial infection by polymerase chain reaction. PCR primers targeted DNA sequences on the chlamydial omp1 or omp2 genes. PCR amplicons were generated from 49.0% of pigs with respiratory disease, from 60.0% of sows with reproductive disorders, from 24.5% of respiratory healthy controls, but from no endocervical swabs from fertile sows. By DNA hybridization, a high prevalence of mixed infections with Chlamydophila abortus and Chlamydia suis in the porcine lung and intestine was found and confirmed by RFLP and nucleotide analysis. Of the omp1-PCR amplicons from endocervical swabs 81.3% were identified as Chlamydophila abortus, indicating an association of this chlamydial species with reproductive disorders in sows. Nucleotide sequence analysis of omp1-amplicons identified as deriving from Chlamydia suis shared a maximum of 82.7% homology with the reference strain S45.
Hoelzle
L E
LE
Institute for Medical Microbiology, Infectious and Epidemic Diseases, Ludwig-Maximilians-University, Munich, Germany.
Steinhausen
G
G
Wittenbrink
M M
MM
D000595:Amino Acid Sequence; D000818:Animals; D001425:Bacterial Outer Membrane Proteins / Q000235:genetics*; D001483:Base Sequence; D002689:Chlamydia / Q000235:genetics*; D002690:Chlamydia Infections / Q000235:genetics / Q000662:veterinary*; D017931:DNA Primers; D005260:Female; D005838:Genotype; D017403:In Situ Hybridization; D008297:Male; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction / Q000662:veterinary*; D012150:Polymorphism, Restriction Fragment Length; D017422:Sequence Analysis, DNA; D013552:Swine; D013553:Swine Diseases / Q000235:genetics*
D001425:Bacterial Outer Membrane Proteins; D017931:DNA Primers; C069991:outer membrane protein, Chlamydia psittaci 6BC