Production of a recombinant fusion protein of Sarcocystis tenella and evaluation of its diagnostic potential in an ELISA.
We have isolated a cDNA clone encoding an antigenic polypeptide of Sarcocystis tenella by screening a cystozoite-derived cDNA library in lambda gt11 with antibodies from sheep infected experimentally with S. tenella. This clone, termed STC29, was subcloned and expressed in the vector pGEX-3X as a soluble fusion protein with glutathione S-transferase having an apparent molecular mass of 46 kDa. Antibody raised against the recombinant fusion protein recognized a native polypeptide of 25 kDa in cystozoites of S. tenella. In an ELISA with sera from experimentally infected sheep, the recombinant STC29 antigen could differentiate infections with S. tenella from those with Sarcocystis arieticanis or Toxoplasma gondii. Hence, the research described here reports the identification of the first recombinant S. tenella antigen that may be useful for standardization of a serological test for the diagnosis of S.tenella infections in sheep.
Mertens
C M
CM
Institut für Parasitologie, Tierärztliche Hochschule Hannover, Germany.
Tenter
A M
AM
Vietmeyer
C
C
Ellis
J T
JT
Johnson
A M
AM
D000595:Amino Acid Sequence; D000818:Animals; D000913:Antibodies, Protozoan / Q000097:blood / Q000276:immunology; D000953:Antigens, Protozoan / Q000235:genetics / Q000276:immunology; D001483:Base Sequence; D015153:Blotting, Western / Q000662:veterinary; D003001:Cloning, Molecular; D016054:DNA, Protozoan / Q000737:chemistry; D004797:Enzyme-Linked Immunosorbent Assay / Q000662:veterinary*; D005260:Female; D015870:Gene Expression; D007106:Immune Sera / Q000276:immunology; D051379:Mice; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction / Q000662:veterinary; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000276:immunology; D012522:Sarcocystis / Q000235:genetics* / Q000276:immunology; D012523:Sarcocystosis / Q000175:diagnosis / Q000276:immunology / Q000662:veterinary*; D016415:Sequence Alignment / Q000662:veterinary; D012756:Sheep; D012757:Sheep Diseases / Q000175:diagnosis* / Q000276:immunology
D000913:Antibodies, Protozoan; D000953:Antigens, Protozoan; D016054:DNA, Protozoan; D007106:Immune Sera; D011993:Recombinant Fusion Proteins