title: Dog cloning with in vivo matured oocytes obtained using electric chemiluminescence immunoassay-predicted ovulation method.
abstract: Radioactive immunoassay (RIA) is a traditional serum hormone assay method, but the application of the method in reproductive studies is limited by the associated radioactivity. The aim of present study was to evaluate the reliability of RIA and to compare its canine serum progesterone concentration determination accuracy to that of the electric chemiluminescence immunoassay (ECLI). In vivo matured oocytes were utilized for canine somatic cell nuclear transfer (SCNT), and serum progesterone levels were assessed to accurately determine ovulation and oocyte maturation. Canine serum progesterone concentrations during both proestrus and estrus were analyzed by RIA and ECLI to determine the ovulation day. Although both methods detected similar progesterone levels before ovulation, the mean progesterone concentration determined using ECLI was significantly higher than of RIA three days before ovulation. Following ovulation, oocytes were collected by surgery, and a lower percentage of mature oocytes were observed using ECLI (39%) as compared to RIA (67%) if 4-8ng/ml of progesterone were used for determination of ovulation. A high percentage of mature oocytes was observed using ECLI when 6-15 ng/mL of progesterone was used for ovulation determination. To determine whether ECLI could be used for canine cloning, six canines were selected as oocyte donors, and two puppies were obtained after SCNT and embryo transfer. In conclusion, compared to the traditional RIA method, the ECLI method is a safe and reliable method for canine cloning.
authors.affiliation: National Institute of Animal Science, RDA, Wanju, Republic of Korea.
authors.forename: Seunghoon
authors.initials: S
authors.lastname: Lee
authors.affiliation: National Institute of Animal Science, RDA, Wanju, Republic of Korea.
authors.forename: Minghui
authors.initials: M
authors.lastname: Zhao
authors.affiliation: National Institute of Animal Science, RDA, Wanju, Republic of Korea.
authors.forename: Jingu
authors.initials: J
authors.lastname: No
authors.affiliation: National Institute of Animal Science, RDA, Wanju, Republic of Korea.
authors.forename: Yoonseok
authors.initials: Y
authors.lastname: Nam
authors.affiliation: National Institute of Animal Science, RDA, Wanju, Republic of Korea.
authors.forename: Gi-Sun
authors.initials: GS
authors.lastname: Im
authors.affiliation: National Institute of Animal Science, RDA, Wanju, Republic of Korea.
authors.forename: Tai-Young
authors.initials: TY
authors.lastname: Hur
mesh_terms: D000818:Animals; D019976:Cloning, Organism / Q000379:methods* / Q000662:veterinary; D004285:Dogs; D004624:Embryo Transfer; D005260:Female; D007118:Immunoassay; D059471:In Vitro Oocyte Maturation Techniques / Q000379:methods*; D008163:Luminescent Measurements; D053652:Nuclear Transfer Techniques; D018587:Oocyte Donation; D009865:Oocytes / Q000502:physiology; D010060:Ovulation; D011247:Pregnancy; D011374:Progesterone / Q000097:blood*; D011863:Radioimmunoassay / Q000379:methods
chemical_list: D011374:Progesterone