title: Molecular analysis of single oocyst of Eimeria by whole genome amplification (WGA) based nested PCR.
abstract: PCR-based molecular tools are widely used for the identification and characterization of protozoa. Here we report the molecular analysis of Eimeria species using combined methods of whole genome amplification (WGA) and nested PCR. Single oocyst of Eimeria stiedai or Eimeriamedia was directly used for random amplification of the genomic DNA with either primer extension preamplification (PEP) or multiple displacement amplification (MDA), and then the WGA product was used as template in nested PCR with species-specific primers for ITS-1, 18S rDNA and 23S rDNA of E. stiedai and E. media. WGA-based PCR was successful for the amplification of these genes from single oocyst. For the species identification of single oocyst isolated from mixed E. stiedai or E. media, the results from WGA-based PCR were exactly in accordance with those from morphological identification, suggesting the availability of this method in molecular analysis of eimerian parasites at the single oocyst level. WGA-based PCR method can also be applied for the identification and genetic characterization of other protists.
authors.affiliation: National Animal Protozoa Laboratory and College of Veterinary Medicine, China Agricultural University, Beijing 100193, China.
authors.forename: Yunzhou
authors.initials: Y
authors.lastname: Wang
authors.affiliation: National Animal Protozoa Laboratory and College of Veterinary Medicine, China Agricultural University, Beijing 100193, China.
authors.forename: Geru
authors.initials: G
authors.lastname: Tao
authors.affiliation: National Animal Protozoa Laboratory and College of Veterinary Medicine, China Agricultural University, Beijing 100193, China.
authors.forename: Yujuan
authors.initials: Y
authors.lastname: Cui
authors.affiliation: National Animal Protozoa Laboratory and College of Veterinary Medicine, China Agricultural University, Beijing 100193, China.
authors.forename: Qiyao
authors.initials: Q
authors.lastname: Lv
authors.affiliation: National Animal Protozoa Laboratory and College of Veterinary Medicine, China Agricultural University, Beijing 100193, China.
authors.forename: Li
authors.initials: L
authors.lastname: Xie
authors.affiliation: National Animal Protozoa Laboratory and College of Veterinary Medicine, China Agricultural University, Beijing 100193, China.
authors.forename: Yuan
authors.initials: Y
authors.lastname: Li
authors.affiliation: National Animal Protozoa Laboratory and College of Veterinary Medicine, China Agricultural University, Beijing 100193, China; Key Laboratory of Animal Epidemiology and Zoonosis of Ministry of Agriculture, China Agricultural University, Beijing 100193, China.
authors.forename: Xun
authors.initials: X
authors.lastname: Suo
authors.affiliation: College of Animal Science and Technology, China Agricultural University, Beijing 100193, China.
authors.forename: Yinghe
authors.initials: Y
authors.lastname: Qin
authors.affiliation: Division of Foodborne, Waterborne and Environmental Diseases, National Center for Emerging and Zoonotic Infectious Diseases, Centers for Disease Control and Prevention, Atlanta, GA 30333, USA.
authors.forename: Lihua
authors.initials: L
authors.lastname: Xiao
authors.affiliation: National Animal Protozoa Laboratory and College of Veterinary Medicine, China Agricultural University, Beijing 100193, China; Key Laboratory of Animal Epidemiology and Zoonosis of Ministry of Agriculture, China Agricultural University, Beijing 100193, China. Electronic address: liuxianyong@cau.edu.cn.
authors.forename: Xianyong
authors.initials: X
authors.lastname: Liu
mesh_terms: D000818:Animals; D003048:Coccidiosis / Q000469:parasitology / Q000662:veterinary; D021901:DNA, Intergenic / Q000235:genetics; D016054:DNA, Protozoan / Q000235:genetics* / Q000302:isolation & purification; D004275:DNA, Ribosomal / Q000235:genetics / Q000302:isolation & purification; D004539:Eimeria / Q000145:classification / Q000235:genetics* / Q000302:isolation & purification; D055106:Genome-Wide Association Study / Q000379:methods*; D058751:Livestock; D033661:Oocysts / Q000145:classification; D016133:Polymerase Chain Reaction; D011200:Poultry; D012337:RNA, Ribosomal, 18S / Q000235:genetics; D012338:RNA, Ribosomal, 23S / Q000235:genetics; D011817:Rabbits
chemical_list: D021901:DNA, Intergenic; D016054:DNA, Protozoan; D004275:DNA, Ribosomal; D012337:RNA, Ribosomal, 18S; D012338:RNA, Ribosomal, 23S
keywords: Eimeria; Morphology; Nested PCR; Single oocyst; Whole genome amplification