Department of Pathology and Laboratory Medicine, NorthShore University HealthSystem, Evanston, Illinois 60201, USA.
Arch Pathol Lab Med. 2013 Apr;137(4):525-30. doi: 10.5858/arpa.2011-0550-OA.
Recently, robotic-assisted laparoscopic prostatectomy has replaced open retropubic radical prostatectomy as the surgical procedure of choice. This less-invasive approach offers many advantages but exposes prostate tissue to longer periods of warm ischemia that may affect subsequent analysis of biomarkers.
To analyze the nucleic acid quality and quantity isolated from open versus laparoscopic prostatectomies.
Nucleic acids were isolated from 10 open-obtained and 10 laparoscopic-obtained tissues stored in our prostate sample repository. Nucleic acid integrity was assessed via electrophoresis and polymerase chain reaction amplification of RNA and DNA targets ranging in size from 125 to 939 base pairs.
The DNA yield, integrity, and polymerase chain reaction amplification were identical between samples obtained from both surgical approaches. The RNA integrity number and yield were similar, as was β-2 microglobulin mRNA amplification up to 652 base pairs. However, 2 of 10 samples (20%) collected robotically showed decreased real-time reverse transcriptase-polymerase chain reaction amplification of prostate-specific antigen messenger RNA, especially with targets larger than 300 base pairs.
Generally, the quality and quantity of nucleic acids isolated from prostate tissue obtained via open or laparoscopic approaches are equivalent, suggesting that procurement of tissues is appropriate from either procedure. However, some loss of reverse transcriptase-polymerase chain reaction amplification of larger RNA targets was noted in the laparoscopic samples; appropriate design of assays to keep amplicon sizes small and the use of internal controls to assess RNA integrity is recommended.
最近，机器人辅助腹腔镜前列腺切除术已取代开放式经耻骨后根治性前列腺切除术，成为首选的手术方法。这种微创方法具有许多优点，但会使前列腺组织暴露在更长时间的温热缺血中，这可能会影响后续生物标志物的分析。
分析从开放式和腹腔镜前列腺切除术获得的核酸的质量和数量。
从我们的前列腺样本库中保存的 10 个开放式和 10 个腹腔镜获得的组织中分离核酸。通过电泳和聚合酶链反应扩增 RNA 和 DNA 靶标（大小范围从 125 到 939 个碱基对）评估核酸的完整性。
从两种手术方法获得的样本的 DNA 产量、完整性和聚合酶链反应扩增结果完全相同。RNA 完整性数量和产量相似，β-2 微球蛋白 mRNA 扩增至 652 个碱基对也是如此。然而，在机器人采集的 10 个样本中的 2 个（20%）中，前列腺特异性抗原信使 RNA 的实时逆转录-聚合酶链反应扩增减少，尤其是靶标大于 300 个碱基对时。
一般来说，从开放式或腹腔镜方法获得的前列腺组织中分离的核酸的质量和数量是等效的，这表明从两种方法中获取组织都是合适的。然而，在腹腔镜样本中观察到一些较大 RNA 靶标的逆转录-聚合酶链反应扩增损失；建议设计适当的检测方法，使扩增片段大小较小，并使用内部对照来评估 RNA 的完整性。
