Giannella C, Zito F A, Colonna F, Paradiso A, Marzullo F, Alaibac M, Schittulli F
Laboratory of Pathology, Institute of Oncology, Bari, Italy.
Eur J Clin Chem Clin Biochem. 1997 Aug;35(8):633-5.
The polymerase chain reaction (PCR) has been successfully employed for the laboratory analyses of genetic and infectious disorders using DNA extracted from paraffin-embedded tissues. However, fixative type and fixation time influenced PCR reactions and in some circumstances amplification fragments could not be efficiently generated. In this study, we determined the effects of three commonly used fixatives including ethanol, formalin and Histochoice, on the PCR amplification of DNA from paraffin-embedded breast cancer tissue. The effect of fixatives and fixation times was measured by the ability of the extracted DNA to serve as a template for the amplification of 280 and 530 base pair DNA fragments. On amplifying DNA, positive reactions were uniformly seen in the ethanol specimens. The next best fixative was Histochoice with positive results almost constantly observed in the PCR reactions performed. Formalin fixation sometimes compromised DNA amplification. Our results are consistent with previous reports investigating the effect of ethanol and formalin fixation on DNA amplification by PCR. Moreover, this is the first study showing that paraffin-embedded tissues fixed with Histochoice can be efficiently used for PCR gene amplification.
聚合酶链反应（PCR）已成功应用于从石蜡包埋组织中提取DNA，对遗传和感染性疾病进行实验室分析。然而，固定剂类型和固定时间会影响PCR反应，在某些情况下无法有效生成扩增片段。在本研究中，我们测定了三种常用固定剂（乙醇、福尔马林和Histochoice）对石蜡包埋乳腺癌组织DNA的PCR扩增的影响。通过提取的DNA作为模板扩增280和530碱基对DNA片段的能力来衡量固定剂和固定时间的影响。扩增DNA时，在乙醇标本中均观察到阳性反应。其次是Histochoice，在进行的PCR反应中几乎始终观察到阳性结果。福尔马林固定有时会影响DNA扩增。我们的结果与先前关于乙醇和福尔马林固定对PCR扩增DNA影响的报道一致。此外，这是第一项表明用Histochoice固定的石蜡包埋组织可有效用于PCR基因扩增的研究。
