Detection and differentiation of Theileria annulata and Theileria parva using macroschizont-derived DNA probes.
A lamda gt11 expression library based on T.annulata-infected cells was screened with an antiserum raised in rabbits against partially purified schizonts of T.annulata. Two clones were detected, sequenced and designated as SA288 and SB288 (Shayan et al., submitted for publication). From the sequences of these two genes oligonucleotide primers were designed for specific amplification of parasite DNA by polymerase chain reaction (PCR). We could show that these genes are of parasitic origin and do occur in all T.annulata stocks tested in the present study. In addition, a target sequence for SA288 could also be identified in T.parva-schizonts. None of them reacted with genomic DNA of different Babesia spp. A third primer pair was designed from the DNA-sequence of a gene encoding for the T.parva-specific casein kinase II-alpha subunit. Using this primer pair, a target sequence could only be detected in T.parva. Taken together, the primers described here can be used as molecular tools in PCR for the detection of Theileria parasites and to distinguish T.annulata from T.parva.
Forschungszentrum Borstel, Germany.
P
P
Shayan
R
R
Biermann
E
E
Schein
J
J
Gerdes
J S
JS
Ahmed
D000469:Alkaline Phosphatase; D000818:Animals; D000913:Antibodies, Protozoan / Q000097:blood*; D000953:Antigens, Protozoan / Q000096:biosynthesis; D002417:Cattle; D017931:DNA Primers; D016054:DNA, Protozoan / Q000032:analysis; D004912:Erythrocytes / Q000469:parasitology; D015723:Gene Library; D007106:Immune Sera; D016133:Polymerase Chain Reaction; D011817:Rabbits; D012469:Salivary Glands / Q000469:parasitology; D016796:Theileria annulata / Q000235:genetics / Q000302:isolation & purification*; D016797:Theileria parva / Q000235:genetics / Q000302:isolation & purification*; D013801:Theileriasis / Q000175:diagnosis* / Q000276:immunology; D013987:Ticks / Q000469:parasitology*
D000913:Antibodies, Protozoan; D000953:Antigens, Protozoan; D017931:DNA Primers; D016054:DNA, Protozoan; D007106:Immune Sera; D000469:Alkaline Phosphatase