Abstract
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.
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REFERENCES
Delhanty JDA: Preimplantation diagnosis. Prenat Diagn 1994;14:1217-1227
Verlinsky Y, Handyside A, Grifo J, Munne S, Cohen J, Liebers I, et al.: Preimplantation diagnosis of genetic and chromosomal disorders. J Assist Reprod Genet 1994;11:236-243
Strom CM, Rechitsky S, Verlinsky Y: Reliability of gender determination using the polymerase chain reaction (PCR) for single cells. J Vitro Fert Embryo Transfer 1991;8:225-229
Akane A, Seki S, Shiono H, Nakamura H, Hasegawa M, Kagawa M, Matsubara K, Nakahori Y, Nagafuchi S, Nakagome Y: Sex determination of forensic samples by dual PCR amplification of an X-Y homologous gene. Forens Sci Int 1992;52:143-148
Liu J, Lissen W, Devroey P, Steirteghem AV, Liebaers I: Amplification of X-and Y-chromosome specific regions from single human blastomeres by polymerase chain reaction for sexing of preimplantation embryos. Hum Reprod 1994;9:716-720
Chong SS, kristjansson K, Cota J, Handyside AH, Hughes MR: Preimplantation prevention of X-linked disease: reliable and rapid sex determination of single human cells by restriction analysis of simultaneously amplified ZFX and ZFY sequences. Hum Mol Genet 1993;2:1187-1191
Waye JS, Willard HF: Chromosome specific alpha satellite DNA: Nucleotide sequence analysis of the 2.0 kilobase-pair repeat from the human X chromosome. Nucleic Acids Res 1985;13:2731-2743
Nakahori Y, Mitani K, Yamada M, Nakagome Y: A human Y-chromosome specific repeated DNA family (DYZ1) consists of a tandem array of pentanucleotides. Nucleic Acids Res 1986;14:7569-7579
Kogan SC, Doherty M, Gitschier J: An improved method for prenatal diagnosis of genetic diseases by analysis of amplified DNA sequences. N Engl J Med 1987;317:985-990
Levinson G, Keyvanfar K., Wu JC, Fugger EF, Fields RA, Harton GL, Palmer FT, Sisson ME, Starr KM, Denninson-Lagos L: DNA-based X-enriched sperm separation as an adjunct to preimplantation genetic testing for prevention of X-linked disease. Hum Reprod 1995;10:979-982
Nakahori Y, Takenaka O, Nakagome Y: A human X-Y homologous region encodes “Amelogenin”. Genomics 1991;9:264-269
Yen PH, Marsh B, Allen E, Tsai SP, Ellison J, Connolly L, Neiswanger K, Shapiro LJ: The human X-linked steroid sulfatase gene and a Y-encoded pseudogene: Evidence for an inversion of the Y chromosome during primate evolution. Cell 1988;55:1123-1135
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Hashiba, T., Sueoka, K., Kuroshima, M. et al. An Accurate and Rapid Gender Determination Assay in Single Cells by the Capillary Polymerase Chain Reaction Method. J Assist Reprod Genet 16, 551–554 (1999). https://doi.org/10.1023/A:1020505406143
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DOI: https://doi.org/10.1023/A:1020505406143