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Preimplantation Genetic Diagnosis for Single Gene Defects
KONG De-wen
2011, 30 (4):
314-318.
Because in overwhelming majority of single gene defects（ＳＧＤ） ｔｈｅ ｇｅｎｅ ｍｕｔａｔｉｏｎｓ ａｒｅ ｈｅｔｅｒｏｚｙｇｏｕｓ， ｉｔ ｉｓ ｎｅｃｅｓｓａｒｙ ｆｏｒ ｍｏｓｔ ｐｒｅ-ｉｍｐｌａｎｔａｔｉｏｎ ｇｅｎｅｔｉｃ ｄｉａｇｎｏｓｉｓ（ＰＧＤ） ｏｆ SGD（ＳＧＤ-ＰＧＤ） ｔｏ ｄｅｓｉｇｎ ｉｎｄｉｖｉｄｕａｌｌｙ． Ｔｈｕｓ， ｔｈｅ ｄｅｓｉｇｎｉｎｇ ｂｅｃｏｍｅｓ ｖｅｒｙ ｉｍｐｏｒｔａｎｔ ｏｎｅ ｏｆ ｔｈｅ ｓｔｅｐｓ ｉｎ ＳＧＤ-ＰＧＤ． Ｓｉｎｃｅ ｔｈｅ ｈｅｒｅｄｉｔｙ ｐａｔｔｅｒｎ（ａｕｔｏｓｏｍａｌ ｄｏｍｉｎａｎｔ ａｎｄ ｒｅｃｅｓｓｉｖｅ， Ｘ-ｌｉｎｋｅｄ ｄｏｍｉｎａｎｔ ａｎｄ ｒｅｃｅｓｓｉｖｅ） ａｎｄ ｔｈｅ ｍｕｔａｔｉｏｎ ｔｙｐｅ（ｐｏｉｎｔ ｍｕｔａｔｉｏｎ， ｌａｒｇｅ ｏｒ ｓｍａｌｌ ｆｒａｇｍｅｎｔ ｉｎｓｅｒｔｉｏｎ ａｎｄ ｄｅｌｅｔｉｏｎ， ｒｅｐｅａｔ ｅｘｐａｎｓｉｏｎ ａｎｄ ｓｏ ｏｎ） ａｒｅ ｄｉｆｆｅｒｅｎｔ ｉｎ ｃａｓｅｓ， ｗｅ ｓｈｏｕｌｄ ｈａｖｅ ｔｈｅ ｃｏｒｒｅｓｐｏｎｄｉｎｇ ｄｅｓｉｇｎｉｎｇ ｔａｃｔｉｃｓ ａｎｄ ｓｔｒａｔｅｇｙ． Ｔｈｒｏｕｇｈ different ｄｅｓｉｇｎing ｅｘａｍｐｌｅｓ， ｔｈｅ ａｕｔｈｏｒ ｗｉｌｌ ｉｎｔｒｏｄｕｃｅ ｔｏ ｒｅａｄｅｒｓ ｄｉｆｆｅｒｅｎｔ ｄｅｓｉｇｎ ｔａｃｔｉｃｓ ａｎｄ ｓｔｒａｔｅｇｉｅｓ． Ｎｏｗａｄａｙｓ ｔｈｅ ｂａｓｉｃ ｔｅｃｈｎｉｑｕｅ ｌｉｎｋ ｏｆ ＳＧＤ-ＰＧＤ ｉｓ ｓｔｉｌｌ ｔｈｅ ａｍｐｌｉｆｉｃａｔｉｏｎ ｆｏｒ ｔａｒｇｅｔ ＤＮＡ ｆｒａｇｍｅｎｔ ｆｒｏｍ ｓｉｎｇｌｅ ｃｅｌｌｓ（ｂｌａｓｔｏｍｅｒｅｓ） ｗｉｔｈ ＤＮＡ ｐｏｌｙｍｅｒａｓｅ ｃｈａｉｎ ｒｅａｃｔｉｏｎ（ＰＣＲ）， ａｎｄ ａｆｔｅｒ ｔｈｅｎ ｔｏ ｄｉａｇｎｏｓｅ ｔｈｅ ｏｂｔａｉｎｅｄ ｔａｒｇｅｔ ｆｒａｇｍｅｎｔ． Ｔｈｅ ｍｏｓｔ ｉｍｐｏｒｔａｎｔ ｓｔｅｐ ｏｆ ｄｅｓｉｇｎｉｎｇ ｉｓ ｄｅｓｉｇｎ ｏｆ ｐｒｉｍｅｒｓ ｔｈａｔ ｗｉｌｌ ｂｅ ｕｓｅｄ ｉｎ ＰＣＲ． Ａｔ ｓａｍｅ ｔｉｍｅｓ ｔｈｅ ｄｅｓｉｇｎｉｎｇ ｓｈｏｕｌｄ ａｌｓｏ ｉｎｃｌｕｄｅ ｔｏ ｓｅｔ ｓｐｅｃｉａｌ ＰＣＲ ｐｒｏｔｏｃｏｌ ｆｏｒ ｓｉｎｇｌｅ ｃｅｌｌ， ｄｅｓｉｇｎ ｒｅｓｔｒｉｃｔｉｏｎ ｅｎｚｙｍｅ ｃｕｔ ｓｉｔｅ（ｓ）[because for most point mutations, restriction fragment length polymorphism（ＲＦＬＰ） method is commonly used for diagnosis], make the analysis pre-estimate of diagnosis result, and determine which primer of the two in the last round of PCR should be fluorescence?-labelled. Any success SGD-PGD requires an authority core of leadership to organize, coordinate, manage and support the whole team, which is also the precondition to serve SGD?-PGD in groups. Moreover, only when there is close coordination among the inheritance epidemiology workers, the paediatric departments of various hospitals, some special disease research institutes and the groups of molecular biology gene diagnosis, SGD?-PGD can be served for various SGDs comprehensively.
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