Evaluation of strain-specific primer sequences from an abortifacient strain of ovine Chlamydophila abortus (Chalmydia psittaci) for the detection of EAE by PCR.
Strain-specific primer sequences derived from the helicase gene of an ovine abortifacient strain (S26/3) of Chlamydophila abortus (Chlamydia psittaci) were evaluated for the diagnosis of enzootic abortion in ewes (EAE) by polymerase chain reaction (PCR). C abortus DNA was amplified from tissues submitted from ovine abortion cases using genus-specific and strain-specific primers in a standard thermal cycler. Amplification was followed by Southern blotting and hybridisation with a strain-specific probe. Real-time PCR was also evaluated using strain-specific primers in a microvolume fluorimeter-based thermal cycler (LightCycler). Detection using both PCR methods was compared with other diagnostic methods against the standard of McCoy cell culture isolation. In this paper we report the application of strain-specific PCR as a fast, sensitive, specific method for the detection of EAE.
Creelan
J L
JL
Department of Agriculture and Rural Development, The Queen's University of Belfast, Stormont, UK.
McCullough
S J
SJ
D000034:Abortion, Veterinary / Q000175:diagnosis* / Q000382:microbiology; D000818:Animals; D000942:Antigens, Bacterial; D001425:Bacterial Outer Membrane Proteins; D015139:Blotting, Southern; D002478:Cells, Cultured; D023501:Chlamydophila / Q000235:genetics / Q000254:growth & development / Q000302:isolation & purification*; D023521:Chlamydophila Infections / Q000175:diagnosis* / Q000382:microbiology / Q000662:veterinary; D017931:DNA Primers; D005260:Female; D005470:Fluorometry; D008565:Membrane Proteins / Q000235:genetics; D016133:Polymerase Chain Reaction / Q000379:methods*; D011247:Pregnancy; D012680:Sensitivity and Specificity; D012756:Sheep; D012757:Sheep Diseases / Q000175:diagnosis* / Q000382:microbiology; D013045:Species Specificity
D000942:Antigens, Bacterial; D001425:Bacterial Outer Membrane Proteins; D017931:DNA Primers; D008565:Membrane Proteins