引物原位标记技术快速检测人类染色体非整倍性.pdfVIP

HEREDITAS (Beijing) 2008 8 , 30(8): 983―990 ISSN 0253-9772 研究报告 DOI: 10.3724/SP.J.1005.2008.00983 朱一剑, 刘涤石, 丁显平 , , 610064 : 染色体数目异常是人类染色体疾病的重要类型, 经常导致胚胎丢失、胎儿流产、婴儿死亡、先天畸形和 神经发育异常等出生缺陷。文章应用引物原位标记(Primed in situ labeling, PRINS)技术快速检测人类染色体非整 倍性, 率先采用更新的非 ddNTP 阻断的多色 PRINS 技术, 对人类外周血淋巴细胞和精子等多种样本进行标记; 然后对不同靶标序列的标记效率及不同荧光色素的发光特点通过实验进行评估, 获得关于 PRINS 技术的多项 反应原理参数, 并筛选标记顺序以获得均一稳定的标记效果, 最后进行临床 FISH 探针与 PRINS 的标记比较实 验。通过实验比较 PRINS 技术与传统 FISH 技术之间的标记特点与差别, 评估 PRINS 的实际应用效果。在 2.5 h 内标记了同一精子核内的多条染色体, 单色以上标记达到 99 ％。同时在人类外周血淋巴细胞中也得到较好的标 记效果。与 FISH 技术相比, PRINS 的这些优点使得它成为诊断染色体非整倍性变异的首选技术。 : 引物原位标记; 人类染色体; 非整倍性 Detecting human chromosome anomalies with primed in situ label- ing (PRINS) ZHU Yi-Jian , LIU Di-Shi , DING Xian-Ping Key Laboratory of Bio-resources and Eco-environment of Ministry of Education, Institute of Medical Genetics, Sichuan University, Chengdu 610064, China Abstract: Numerical chromosome anomaly was one of the most important kinds of human chromosme diseases by induc- ing pregnancy loss, miscarriage, infant death, congenital malformations and nerve damage. The present study was to estab- lish a rapid, reliable and reasonable multicolor primed in situ labeling (PRINS) protocol for diagnosing numerical anomaly in human chromosome. First, nuclei of cultured lymphocytes and sperms were labeled with the method of PRINS, and then nuclei of cultured lymphocytes, sperms and other specimen were labeled with the method of updated non-ddNTP-blocking multicolor PRINS technique. The labeling effect of different target sequences and the feature of different fluorochromes were evaluated by experiment. Meanwhile, se