Genotyping of Chlamydophila psittaci by real-time PCR and high-resolution melt analysis.
Human infection with Chlamydophila (Chlamydia) psittaci can lead to psittacosis, a disease that occasionally results in severe pneumonia and other medical complications. C. psittaci is currently grouped into seven avian genotypes: A through F and E/B. Serological testing, outer membrane protein A (ompA) gene sequencing, and restriction fragment length polymorphism analysis are currently used for distinguishing these genotypes. Although accurate, these methods are time-consuming and require multiple confirmatory tests. By targeting the ompA gene, a real-time PCR assay has been developed to rapidly detect and genotype C. psittaci by light-upon-extension chemistry and high-resolution melt analysis. Using this assay, we screened 169 animal specimens; 98 were positive for C. psittaci (71.4% genotype A, 3.1% genotype B, 4.1% genotype E, and 21.4% unable to be typed). This test may provide insight into the distribution of each genotype among specific hosts and provide epidemiological and epizootiological data in human and mammalian/avian cases. This diagnostic assay may also have veterinary applications during chlamydial outbreaks, particularly with respect to identifying the sources and tracking the movements of a particular genotype when multiple animal facilities are affected.
Mitchell
Stephanie L
SL
Respiratory Disease Branch, Centers for Disease Control and Prevention, 1600 Clifton Rd. NE, MS G-03, Atlanta, GA 30333, USA.
Wolff
Bernard J
BJ
Thacker
W Lanier
WL
Ciembor
Paula G
PG
Gregory
Christopher R
CR
Everett
Karin D E
KD
Ritchie
Branson W
BW
Winchell
Jonas M
JM
D000818:Animals; D001425:Bacterial Outer Membrane Proteins / Q000235:genetics; D001717:Birds; D002691:Chlamydophila psittaci / Q000145:classification* / Q000235:genetics* / Q000302:isolation & purification; D017931:DNA Primers / Q000235:genetics; D004269:DNA, Bacterial / Q000235:genetics*; D006801:Humans; D016133:Polymerase Chain Reaction / Q000379:methods*; D009956:Psittacosis / Q000175:diagnosis*; D044366:Transition Temperature
D001425:Bacterial Outer Membrane Proteins; D017931:DNA Primers; D004269:DNA, Bacterial; C078167:OMPA outer membrane proteins