Jiao Zhe-xu, Zhuang Guang-lun, Zhou Can-quan, Zhang Min-fang, Li Li-lin
Reproductive Medical Center, First Affiliated Hospital of Sun Yat-Sen University, Guangzhou, Guangdong, PR China.
Zhonghua Yi Xue Yi Chuan Xue Za Zhi. 2003 Feb;20(1):64-5.
Using nested polymerase chain reaction (PCR) to perform preimplantation gender diagnosis.
One (or two) lymphocyte and blastomere (n=50/group) were collected and prepared under the following conditions: (1) water only (H(2)O); (2) freeze-thaw liquid nitrogen, then boiling; (3) potassium hydroxide/dithiotheriol, heated to 65 degree centigrade, followed by acid neutralization (KOH). Cells were analyzed by PCR using nested primers amplification with amelogenin gene.
The amplification rate and allele dropout (ADO) rate for male lymphocytes by the three methods were 83%, 94%, 95% and 24%, 12%, 4%, respectively. Using two cells per reaction did not increase the amplification rate for the KOH method.
The KOH method for DNA preparation is superior to the other methods evaluated. Dual blastomere biopsy and independent blastomere analysis may improve preimplantation diagnostic reliability.
采用巢式聚合酶链反应（PCR）进行植入前性别诊断。
收集一（或两）个淋巴细胞和卵裂球（每组n = 50），并在以下条件下制备：（1）仅用水（H₂O）；（2）液氮冻融，然后煮沸；（3）氢氧化钾/二硫苏糖醇，加热至65摄氏度，随后进行酸中和（KOH）。使用巢式引物扩增牙釉蛋白基因，通过PCR对细胞进行分析。
三种方法对男性淋巴细胞的扩增率和等位基因脱扣（ADO）率分别为83%、94%、95%和24%、12%、4%。每个反应使用两个细胞并未提高KOH法的扩增率。
KOH法制备DNA优于所评估的其他方法。双卵裂球活检和独立卵裂球分析可能提高植入前诊断的可靠性。
