Differential diagnosis of five avian Eimeria species by polymerase chain reaction using primers derived from the internal transcribed spacer 1 (ITS-1) sequence.
Chicken coccidia are protozoan parasites of the genus Eimeria. They cause economical losses in the poultry industry globally. The various species can be distinguished on the basis of the morphology of the oocysts and parasitic site in intestine, but these criteria sometimes are unreliable. Therefore, a species-specific polymerase chain reaction (PCR) was developed. Based on variable sequence regions, specific primers were constructed for the differentiation of five Eimeria species (Eimeria acervulina, E. brunette, E. maxima, E. necatrix, and E. tenella). PCR products were amplified from coccidian vaccine (coccivac-D and coccivac-B) and E. tenella and were subsequently sequenced. Similarities of the five species sequences between the vaccines and Genbank were 94-100%. Analysis of the E. tenella internal transcribed spacer 1 (ITS-1) partial sequence from Taiwan and from Genbank indicated that the similarity was 99.6%. The PCR sensitivity test of E. tenella in Taiwan is 50 oocysts. The five sets of primers will not amplify any non-specific bands of the chicken genome or its intestinal contents. Therefore, the five sets of specifically designed primers are guaranteed to be useful for differential diagnosis of avian coccidiosis caused by Eimeria spp.
Su
Yao-Chi
YC
Department of Veterinary Medicine, National Chiayi University, 300 University Road, Chiayi City 600, Taiwan, ROC.
Fei
Andrew Chang-Young
AC
Tsai
Fang-Mei
FM
D000818:Animals; D001483:Base Sequence; D002645:Chickens / Q000469:parasitology*; D003048:Coccidiosis / Q000175:diagnosis / Q000469:parasitology / Q000662:veterinary*; D017931:DNA Primers; D021901:DNA, Intergenic / Q000737:chemistry* / Q000235:genetics; D003937:Diagnosis, Differential; D004539:Eimeria / Q000145:classification / Q000235:genetics* / Q000302:isolation & purification; D008969:Molecular Sequence Data; D010802:Phylogeny; D016133:Polymerase Chain Reaction / Q000379:methods / Q000662:veterinary; D011201:Poultry Diseases / Q000175:diagnosis* / Q000469:parasitology; D016052:Protozoan Vaccines; D016053:RNA, Protozoan / Q000032:analysis; D012335:RNA, Ribosomal / Q000032:analysis; D012689:Sequence Homology, Nucleic Acid; D013045:Species Specificity
D017931:DNA Primers; D021901:DNA, Intergenic; D016052:Protozoan Vaccines; D016053:RNA, Protozoan; D012335:RNA, Ribosomal