title: Rapid sex chromosomal chimerism analysis in heterosexual twin female calves by Loop-mediated Isothermal Amplification.
abstract: We attempted to apply an embryo sexing kit with Loop-mediated Isothermal Amplification (LAMP) to sex chromosomal chimerism analysis in heterosexual twin female calves. Peripheral blood was used for the amplification of male-specific DNA, derived from XY leukocytes. When blood samples were diluted 1:1000 in LAMP reaction mixture, hemoglobin or blood coagulation did not influence the turbidity measurement of the reaction mixture for detection of amplified DNA. This procedure detected the existence of XY leukocytes of 0.01% in female blood. Furthermore, all heterosexual twin female calves, bearing sex chromosomal chimerism based on karyotyping and PCR, showed male-specific DNA from peripheral blood by LAMP. These results indicated that the embryo sexing kit with LAMP was available for sensitive detection of sex chromosomal chimerism. This procedure made it possible to detect easily Y-chromosome specific DNA in a short interval compared with PCR, and was convenient for field application of freemartin diagnosis.
authors.affiliation: Hokkaido Animal Research Center, Hokkaido 081-0038, Japan. h.hirayama@agri.pref.hokkaido.jp
authors.forename: Hiroki
authors.initials: H
authors.lastname: Hirayama
authors.forename: Seiji
authors.initials: S
authors.lastname: Katagiri
authors.forename: Soichi
authors.initials: S
authors.lastname: Kageyama
authors.forename: Akira
authors.initials: A
authors.lastname: Minamihashi
authors.forename: Satoru
authors.initials: S
authors.lastname: Moriyasu
authors.forename: Ken
authors.initials: K
authors.lastname: Sawai
authors.forename: Sadao
authors.initials: S
authors.lastname: Onoe
authors.forename: Yoshiyuki
authors.initials: Y
authors.lastname: Takahashi
mesh_terms: D000818:Animals; D002417:Cattle; D002678:Chimera / Q000235:genetics*; D004252:DNA Mutational Analysis / Q000379:methods*; D005260:Female; D009391:Nephelometry and Turbidimetry; D021141:Nucleic Acid Amplification Techniques / Q000379:methods*; D011247:Pregnancy; D011296:Prenatal Diagnosis / Q000379:methods*; D012680:Sensitivity and Specificity; D012729:Sex Chromosome Aberrations; D013696:Temperature; D014427:Twins; D014960:X Chromosome; D014998:Y Chromosome