Improved sensitivity of PCR for Chlamydophila using pmp genes.
Primers targeting the conserved pmp gene family of Chlamydophila abortus were evaluated for their ability to improve the polymerase chain reaction (PCR) sensitivity. In purified DNA, specific pmp primers (named CpsiA and CpsiB) allowed at least a 10-fold increase of the PCR sensitivity compared to the specific ompA primers for C. abortus, but also for C. psittaci and C. caviae strains. No amplification was observed on C. felis, C. pecorum, C. pneumoniae and Chlamydia trachomatis strains. Tested on contaminated specimens such as genital swabs, the PCR sensitivity observed with CpsiA/CpsiB was also better than with the ompA primers. This study demonstrated that these specific pmp primers could serve as valuable, sensitive and common tools for a specific Chlamydophila diagnosis in ruminant, avian and human diseases. Digestion by AluI of the CpsiA/CpsiB fragments allowed a specific discrimination of the strains in function of their hosts and/or their serotypes.
Laroucau
K
K
Unité de Pathologie Infectieuse et Immunologie, INRA-Centre de Tours, 37380 Nouzilly, France.
Souriau
A
A
Rodolakis
A
A
D000818:Animals; D001425:Bacterial Outer Membrane Proteins / Q000235:genetics*; D001483:Base Sequence; D023501:Chlamydophila / Q000235:genetics* / Q000302:isolation & purification; D017931:DNA Primers; D005260:Female; D005784:Gene Amplification; D006801:Humans; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction / Q000379:methods / Q000592:standards / Q000662:veterinary*; D012680:Sensitivity and Specificity; D012756:Sheep; D014621:Vagina / Q000382:microbiology
D001425:Bacterial Outer Membrane Proteins; D017931:DNA Primers