PCR detection of Coxiella burnetii from different clinical specimens, especially bovine milk, on the basis of DNA preparation with a silica matrix.
For PCR detection of Coxiella burnetii in various clinical specimens we developed a sample preparation method in which silica binding of DNA was used. This method was found to be fast, easily performed with large numbers of samples, and equally sensitive for all of the specimens tested (livers, spleens, placentas, heart valves, milk, blood). The DNA preparation method described here can also be used as an initial step in any PCR-based examination of specimens. The procedure was tested with more than 600 milk samples, which were taken from 21 cows that were seropositive for C. burnetii and reportedly had fertility problems (and therefore were suspected of shedding the agent through milk intermittently or continuously). Of the 21 cows tested, 6 were shedding C. burnetii through milk. Altogether, C. burnetii DNA was detected in 6% of the samples. There was no correlation between the shedding pattern and the serological results.
Lorenz
H
H
Institute for Hygiene and Infectious Diseases of Animals, Justus-Liebig University, Giessen, Germany.
Jäger
C
C
Willems
H
H
Baljer
G
G
D000818:Animals; D002417:Cattle; D002418:Cattle Diseases / Q000175:diagnosis* / Q000382:microbiology; D002460:Cell Line; D002522:Chlorocebus aethiops; D016997:Coxiella burnetii / Q000235:genetics / Q000302:isolation & purification*; D004269:DNA, Bacterial / Q000235:genetics / Q000302:isolation & purification*; D005260:Female; D006351:Heart Valves / Q000382:microbiology; D008099:Liver / Q000382:microbiology; D008892:Milk / Q000382:microbiology*; D010920:Placenta / Q000382:microbiology; D016133:Polymerase Chain Reaction / Q000379:methods; D011247:Pregnancy; D011778:Q Fever / Q000175:diagnosis / Q000662:veterinary*; D012680:Sensitivity and Specificity; D012756:Sheep; D012757:Sheep Diseases / Q000175:diagnosis / Q000382:microbiology; D012822:Silicon Dioxide; D013154:Spleen / Q000382:microbiology; D013552:Swine; D013553:Swine Diseases / Q000175:diagnosis / Q000382:microbiology
D004269:DNA, Bacterial; D012822:Silicon Dioxide