Detection of Coxiella burnetii in placenta and abortion samples from British ruminants using real-time PCR.
A real-time PCR was developed to detect Coxiella burnetii (the cause of Q fever) in ruminant placentas and aborted fetuses. Primer and probe sets previously developed for human tissue studies were used to target the insertion sequence IS1111 gene for C burnetii. The assay was highly sensitive, with a limit of detection of 10 copies of template, theoretically equating to a single bacterium, and did not cross-react with a panel of other bacteria. To determine sensitivity on field samples submitted for the diagnosis of abortion, results using the IS1111 PCR assay were compared with a com1 PCR assay. When applied to ruminant abortion material, including placental cotyledons and fetal samples, the IS1111 and com1 assays yielded positive results in 23 (25 per cent) of 93 and 19 (20 per cent) of 93 samples, respectively. One infected goat herd was monitored for 31 months: 57 (92 per cent) of 62 placental cotyledon samples from aborting and non-aborting goats, and 10 (30 per cent) of 33 fetal samples were positive by the IS1111 PCR assay.
Veterinary Laboratories Agency (VLA) - Weybridge, New Haw, Addlestone, Surrey.
R M
RM
Jones
D F
DF
Twomey
S
S
Hannon
J
J
Errington
G C
GC
Pritchard
J
J
Sawyer
D037881:Aborted Fetus / Q000382:microbiology*; D000034:Abortion, Veterinary / Q000175:diagnosis / Q000382:microbiology*; D000818:Animals; D002417:Cattle; D002418:Cattle Diseases / Q000382:microbiology; D016997:Coxiella burnetii / Q000235:genetics / Q000302:isolation & purification*; D005260:Female; D015511:Goat Diseases / Q000382:microbiology; D006041:Goats; D010920:Placenta / Q000382:microbiology*; D016133:Polymerase Chain Reaction / Q000379:methods / Q000662:veterinary*; D011247:Pregnancy; D011778:Q Fever / Q000175:diagnosis / Q000662:veterinary*; D012680:Sensitivity and Specificity; D012756:Sheep; D012757:Sheep Diseases / Q000382:microbiology