Paulasova P, Andréo B, Diblik J, Macek M, Pellestor F
Laboratory of Assisted Reproduction, Motol Hospital, Vuvalu 84, 150 06 Praha 5, Czech Republic.
Mol Hum Reprod. 2004 Jun;10(6):467-72. doi: 10.1093/molehr/gah061. Epub 2004 Apr 20.
Peptide nucleic acids (PNA) are synthetic DNA mimics based on an uncharged polyamide backbone, which hybridize with complementary DNA with high affinity and specificity. PNA have recently become recognized as efficient tools for in situ chromosomal identification. In the present study, this new approach has been tried on isolated human oocytes, polar bodies and blastomeres. Using centromeric PNA probes specific for chromosomes 1, 4, 9, 16, X and Y, we tested multicolour labelling PNA reaction on 27 oocytes and 23 blastomeres. Sequential PNA hybridization was performed on five oocytes and combined PNA and fluorescence in situ hybridization (FISH) reactions on two oocytes. Both the rates and the types of abnormalities observed are in agreement with results from previous FISH studies. This preliminary study indicates that PNA probes allow a reliable chromosomal analysis in isolated human oocytes and blastomeres and consequently might provide an interesting adjunct to FISH for diagnostic analysis.
肽核酸(PNA)是基于不带电荷的聚酰胺主链的合成DNA模拟物,它能以高亲和力和特异性与互补DNA杂交。PNA最近已被公认为是用于原位染色体鉴定的有效工具。在本研究中,这种新方法已在分离出的人类卵母细胞、极体和卵裂球上进行了尝试。使用针对1、4、9、16、X和Y染色体的着丝粒PNA探针,我们在27个卵母细胞和23个卵裂球上测试了多色标记PNA反应。对5个卵母细胞进行了连续PNA杂交,并对2个卵母细胞进行了PNA与荧光原位杂交(FISH)反应的联合检测。观察到的异常发生率和类型均与先前FISH研究的结果一致。这项初步研究表明,PNA探针可对分离出的人类卵母细胞和卵裂球进行可靠的染色体分析,因此可能为FISH诊断分析提供有趣的辅助手段。