Strategy for detection and differentiation of Coxiella burnetii strains using the polymerase chain reaction.
A method for the rapid detection of Coxiella burnetii and differentiation between strains that cause endocarditis and those that cause acute Q fever is based on the observation that the different strains contain unique plasmid sequences. This method employs the polymerase chain reaction (PCR) and requires knowledge of specific DNA sequences in the region (target) of DNA to be amplified. To detect and differentiate between C. burnetii isolates, two sets of primers are required. The first set was derived from a fragment of plasmid QpH1 which has been detected in all C. burnetii isolates. A second PCR reaction was conducted using primers specific for DNA sequences that are shared only by QpRS plasmid-containing strains of C. burnetii. The first reaction detects the presence of C. burnetii. The second PCR is necessary to determine whether the isolate contains DNA sequences associated with strains causing chronic disease. These procedures detect as few as one to ten organisms.
Mallavia
L P
LP
Washington State University, Pullman 99164.
Whiting
L L
LL
Minnick
M F
MF
Heinzen
R
R
Reschke
D
D
Foreman
M
M
Baca
O G
OG
Frazier
M E
ME
D003381:Coxiella / Q000235:genetics / Q000302:isolation & purification*; D004269:DNA, Bacterial / Q000032:analysis*; D016133:Polymerase Chain Reaction
D004269:DNA, Bacterial