An accurate and rapid gender determination assay in single cells by the capillary polymerase chain reaction method

Tsuyoshi Hashiba, Kou Sueoka, Masako Kuroshima, Hironori Asada, Naoaki Kuji, Yasunori Yoshimura

Research output: Contribution to journalArticle

2 Citations (Scopus)

Abstract

Purpose: In preimplantation genetic diagnosis (PGD), a rapid and accurate assay has been required. We have therefore developed a capillary palymerase 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 DXZI and a 154-bp DYZI 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.

Original languageEnglish
Pages (from-to)551-554
Number of pages4
JournalJournal of Assisted Reproduction and Genetics
Volume16
Issue number10
DOIs
Publication statusPublished - 1999 Nov 22

Keywords

  • Capillary PCR
  • DXZI
  • DYZI
  • Gender determination
  • Preimplantation genetic diagnosis

ASJC Scopus subject areas

  • Reproductive Medicine
  • Genetics
  • Obstetrics and Gynaecology
  • Developmental Biology
  • Genetics(clinical)

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