回收固定在显微镜载玻片上的单个人类卵裂球，用于通过各种类型的聚合酶链反应进行性别鉴定和特定突变的诊断。

Recycling of a single human blastomere fixed on a microscopic slide for sexing and diagnosis of specific mutations by various types of polymerase chain reaction.

作者信息

He Z Y, Liu H C, Mele C A, Veeck L L, Davis O, Rosenwaks Z

机构信息

Center for Reproductive Medicine and Infertility, Weill Medical College of Cornell University, New York, New York, USA.

出版信息

Fertil Steril. 1999 Aug;72(2):341-8. doi: 10.1016/s0015-0282(99)00241-1.

DOI:10.1016/s0015-0282(99)00241-1

PMID:10439008

Abstract

OBJECTIVE

To investigate the suitability of recycling single blastomeres to assess multiple genetic variables for preimplantation genetic diagnosis.

DESIGN

Prospective randomized study.

SETTING

An academic medical center.

PATIENT(S): Patients undergoing IVF-ET.

INTERVENTION(S): Blastomeres were disaggregated from donated embryos obtained from patients.

MAIN OUTCOME MEASURE(S): Polymerase chain reaction (PCR) amplification products.

RESULT(S): Fifty-eight blastomeres individually fixed on slides were separated into four groups. Sequential PCRs (group I, n = 30), primed in situ labeling (PRINS) before five sequential PCRs (group II, n = 10), staining with hematoxylin before performing five sequential PCRs (group III, n = 11) and preamplification of whole DNAs by degenerate oligonucleotide primer (DOP) before performing PCR were executed. The amplification efficiencies of five sequential PCRs were 100%, 100%, 96.6%, 83.3%, 56.7% for group I; 100% 100%, 100%, 80%, 40% for group II; 54.5%, 36.4%, 18.2%, 9.1% for group III; and 100%, 100%, 100%, 100%, 100% for group IV.

CONCLUSION(S): Blastomeres fixed for PRINS can be recycled for PCR to obtain more genetic information. Hematoxylin staining appears to increase the incidence of failed amplification. Preamplification of whole genomic DNAs by DOP-PCR appears to facilitate diagnosis with high efficiency.

摘要

目的

研究回收单个卵裂球以评估植入前基因诊断中多个基因变量的适用性。

设计

前瞻性随机研究。

地点

一家学术医疗中心。

患者

接受体外受精-胚胎移植的患者。

干预措施

从患者捐赠的胚胎中分离出卵裂球。

主要观察指标

聚合酶链反应（PCR）扩增产物。

结果

58个单独固定在载玻片上的卵裂球被分为四组。分别进行了连续PCR（第一组，n = 30）、在连续五次PCR之前进行引物原位标记（PRINS）（第二组，n = 10）、在进行连续五次PCR之前用苏木精染色（第三组，n = 11）以及在进行PCR之前通过简并寡核苷酸引物（DOP）对全基因组DNA进行预扩增（第四组）。第一组连续五次PCR的扩增效率分别为100%、100%、96.6%、83.3%、56.7%；第二组为100%、100%、100%、80%、40%；第三组为54.5%、36.4%、18.2%、9.1%；第四组为100%、100%、100%、100%、100%。