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KMID : 0358219960230020201
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Korean Journal of Fertility and Sterility
1996 Volume.23 No. 2 p.201 ~ p.206
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Analysis of SRY and ZP3 Gene in Single Blastomere of Preimplantation Human Embryos using Polymerase Chain Reaction
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이정헌
윤현수/이동률/김해정/채영규/조정현/노성일
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Abstract
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When there is a risk of inherited disease, preimplantation genetic diagnosis gives couples an advantage to avoid having a child with the genetic disease. In order to evaluate the efficiency of polymerase chain reaction (PCR) at the single cell
level in
human IVF-ET program, we analysed the testis-determining gene (SRY) on the short arm of human Y chromosome and ZP3 gene on chromosome 7 in single blastomere of 6-cell to 10-cell embryos. Ninety-nine blastomeres from 49 embryos were isolated by
micromanipulation or enzymatic digestion at 3 days after in vitro fertilization. Isolated blastomeres were analysed by multiplex nested PCR. SRY-amplified products (254bp) and ZP3-amplified products (177bp) were identified, and further analysed
by
restriction enzyme PvuII. The total efficiency of PCR amplification was 88% (87/99), but the efficiency of PCR amplification for blastomeres derived from 2PN embryos was 100%. The simultaneous analysis of SRY and ZP2 gene can be performed within
8
hours, and it was possible to discriminate between blastomeres derived from male and female embryos. These results represent the possibility for diagnosis of X-linked recessive diseases for single gene disorders in preimplantation embryos by
multiplex
nested PCR.
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