A pmp genes-based PCR as a valuable tool for the diagnosis of avian chlamydiosis.
In a previous study we described the use of a new set of PCR primers (CpsiA/CpsiB) specific of the conserved pmp-family genes of Chlamydophila abortus as an efficient tool for the detection of these bacteria in ruminants including also preliminary results on avian strains. In this work, the use of this set of primers was extended to representative strains of the six major avian serovars (serovars A-F) and to field isolates of C. psittaci. For all the studied representative strains, using purified genomic DNA as a template, CpsiA/CpsiB primers allowed, as observed for C. abortus, a minimal 10-fold PCR signal increase compared to the one observed with ompA specific primers. In comparison to primers targeting the 16S-23S rRNA intergenic spacer, similar or increased sensitivity was observed depending on the strain. All the field isolates were amplified with CpsiA/CpsiB primers. On clinical samples, our primers are the best among those tested for detection of C. psittaci by simple conventional PCR. RFLP experiments performed using PCR fragments amplified with the CpsiA/CpsiB primers gave promising results demonstrating that these primers may provide an interesting tool for molecular typing when the bacterium cannot be grown from pathological samples.
Laroucau
Karine
K
French Food Safety Agency, Bacterial Zoonosis Unit, Maisons-Alfort, France. k.laroucau@afssa.fr
Trichereau
Alain
A
Vorimore
Fabien
F
Mahé
Anne-Marie
AM
D000818:Animals; D001425:Bacterial Outer Membrane Proteins / Q000737:chemistry / Q000235:genetics; D001715:Bird Diseases / Q000175:diagnosis / Q000382:microbiology*; D001717:Birds; D002691:Chlamydophila psittaci / Q000235:genetics / Q000302:isolation & purification*; D004269:DNA, Bacterial / Q000737:chemistry / Q000235:genetics; D016133:Polymerase Chain Reaction / Q000662:veterinary*; D012150:Polymorphism, Restriction Fragment Length; D009956:Psittacosis / Q000175:diagnosis / Q000382:microbiology / Q000662:veterinary*; D012680:Sensitivity and Specificity
D001425:Bacterial Outer Membrane Proteins; D004269:DNA, Bacterial