PURPOSE: In preimplantation genetic diagnosis (PGD), a rapid and accurate assay has been required. We have therefore developed a capillary polymerase chain reaction (PCR) method using rapid thermal cycling programs to determine the gender of single amniocytes. METHODS: Single amniocytes from each amniotic fluid sample were isolated by micromanipulation and their gender was determined by a multiplex PCR assay in a capillary tube, using primers that amplify a 308-bp DXZ1 and a 154-bp DYZ1 repeat sequence on the X and Y chromosomes, respectively. RESULTS: All four thermal cycling programs, which took 180, 150, 120, and 90 min, were 100% accurate in diagnosing the gender of single amniocytes. No DNA contamination was observed in any samples. CONCLUSIONS: The multiplex PCR assay was rapid and accurate in diagnosing gender in single cells and may be clinically applicable in PGD.
PURPOSE: In preimplantation genetic diagnosis (PGD), a rapid and accurate assay has been required. We have therefore developed a capillary polymerase chain reaction (PCR) method using rapid thermal cycling programs to determine the gender of single amniocytes. METHODS: Single amniocytes from each amniotic fluid sample were isolated by micromanipulation and their gender was determined by a multiplex PCR assay in a capillary tube, using primers that amplify a 308-bp DXZ1 and a 154-bp DYZ1 repeat sequence on the X and Y chromosomes, respectively. RESULTS: All four thermal cycling programs, which took 180, 150, 120, and 90 min, were 100% accurate in diagnosing the gender of single amniocytes. No DNA contamination was observed in any samples. CONCLUSIONS: The multiplex PCR assay was rapid and accurate in diagnosing gender in single cells and may be clinically applicable in PGD.
Authors: A Akane; S Seki; H Shiono; H Nakamura; M Hasegawa; M Kagawa; K Matsubara; Y Nakahori; S Nagafuchi; Y Nakagome Journal: Forensic Sci Int Date: 1992-01 Impact factor: 2.395
Authors: Y Verlinsky; A Handyside; J Grifo; S Munné; J Cohen; I Liebers; G Levinson; N Arnheim; M Hughes; J Delhanty Journal: J Assist Reprod Genet Date: 1994-05 Impact factor: 3.412
Authors: G Levinson; K Keyvanfar; J C Wu; E F Fugger; R A Fields; G L Harton; F T Palmer; M E Sisson; K M Starr; L Dennison-Lagos Journal: Hum Reprod Date: 1995-04 Impact factor: 6.918