Pellestor F, Malki S, Andréo B, Lefort G
CNRS-UPR 1142, Montpellier, France.
Chromosome Res. 2002;10(5):359-67. doi: 10.1023/a:1016845517798.
We report a new multicolor PRINS procedure for chromosome identification on human sperm. Based on the direct in-situ mixing of the colors of the fluorochromes (FITC, TRITC, Cascade Blue) incorporated in sequential PRINS reactions, this method facilitates rapid distinct labeling of 3 or 4 chromosomes. Each PRINS reaction consists of a unique 4 minute step for annealing and elongation. The method was successfully tested on lymphocytes and spermatozoa. Estimates of disomy were performed for chromosomes 7, 9 and 16 on sperm samples from 2 healthy donors. There was no significant difference between the disomy rates obtained with the conventional two-color PRINS technique and this new three-color procedure. By simplifying the multicolor PRINS protocol, this new protocol should facilitate the use and adaptation of PRINS to various cytogenetic applications.
我们报告了一种用于人类精子染色体鉴定的新型多色引物原位标记法(PRINS)。基于连续PRINS反应中掺入的荧光染料(异硫氰酸荧光素、四甲基异硫氰酸罗丹明、叠氮蓝)颜色的直接原位混合,该方法有助于对3条或4条染色体进行快速清晰的标记。每个PRINS反应包括一个独特的4分钟退火和延伸步骤。该方法已在淋巴细胞和精子上成功测试。对来自2名健康供体的精子样本中的7号、9号和16号染色体进行了二体性估计。传统双色PRINS技术与这种新的三色方法获得的二体率之间没有显著差异。通过简化多色PRINS方案,这种新方案应有助于PRINS在各种细胞遗传学应用中的使用和应用。