Detection of Babesia bovis using a DIG-labeled DNA probe.
A plasmid DNA containing the inserted Babesia bovis cDNA clone designated c51A was used to prepare a DNA probe for B. bovis. The purified 0.6 kb specific DNA fragment was labeled by DIG DNA labeling. After denaturation, the probe was hybridised with the blotted target DNA extracted from bovine red blood cells infected with B. bovis or other protozoa or bovine red blood cells plus extra white blood cells. It was found that the probe produced from purified 0.6 kb DNA fragment could detect sample of B. bovis DNA equivalent to 0.015 microliter of 10% whole infected blood. Compared to the same DNA fragment labeled with photobiotin, this new probe is more sensitive giving a darker hybridization signal, a lighter hybridization background and without any non-specific reactions. These results indicated that this DIG-labeled B. bovis C5lA probe could provide a sensitive and specific method to diagnose clinically suspected B. bovis infections and distinguish B. bovis DNA from other haemoprotozoan infections.
Wu
J
J
Animal Quarantine Institute, Ministry of Agriculture, Qingdao, China.
Xie
M
M
Zhang
X
X
Wu
H
H
Wang
Z
Z
Xie
H
H
Du
Y
Y
Pong
X
X
D000345:Affinity Labels; D000818:Animals; D001386:Azides; D016793:Babesia bovis / Q000235:genetics / Q000302:isolation & purification*; D001404:Babesiosis / Q000175:diagnosis*; D001710:Biotin / Q000031:analogs & derivatives; D002417:Cattle; D002418:Cattle Diseases / Q000175:diagnosis* / Q000469:parasitology; D015342:DNA Probes; D016054:DNA, Protozoan / Q000032:analysis*; D004076:Digoxigenin; D004587:Electrophoresis, Agar Gel; D005821:Genetic Techniques; D009693:Nucleic Acid Hybridization; D012680:Sensitivity and Specificity
D000345:Affinity Labels; D001386:Azides; D015342:DNA Probes; D016054:DNA, Protozoan; D001710:Biotin; C045549:photobiotin; D004076:Digoxigenin