Lei Benson U W, Yamada Miko, Hoang Van L T, Belt Paul J, Moore Mark H, Lin Lynlee L, Flewell-Smith Ross, Dang Nhung, Tomihara Shoko, Prow Tarl W
Future Industries Institute, University of South Australia; Faculty of Medicine, University of Queensland.
Future Industries Institute, University of South Australia.
J Vis Exp. 2019 Feb 21(144). doi: 10.3791/58614.
Conventional skin biopsy limits the clinical research that involves cosmetically sensitive areas or pediatric applications due to its invasiveness. Here, we describe the protocol for using an absorbent microneedle-based device, absorbent microbiopsy, for minimally invasive sampling of skin and blood mixture. Our goal is to help facilitate rapid progress in clinical research, the establishment of biomarkers for skin disease and reducing the risk for clinical research participants. In contrast to conventional skin biopsy techniques, the absorbent microbiopsy can be performed within seconds and does not require intensive training due to its simple design. In this report, we describe the use of absorbent microbiopsy, including loading and application, on a volunteer. Then, we show how to isolate RNA from the absorbed sample. Finally, we demonstrate the use of quantitative reverse transcription PCR (RT-qPCR) to quantify mRNA expression levels of both blood (CD3E and CD19) and skin (KRT14 and TYR). The methods that we describe utilize off the shelf kits and reagents. This protocol offers a minimally invasive approach for simultaneous sampling of skin and blood within the same absorbent microbiopsy matrix. We have found human ethics committees, clinicians and volunteers to be supportive of this approach to dermatological research.
传统的皮肤活检因其侵入性，限制了涉及美容敏感区域或儿科应用的临床研究。在此，我们描述了一种基于吸收性微针装置（吸收性微生物活检）的方案，用于对皮肤和血液混合物进行微创采样。我们的目标是帮助推动临床研究的快速进展、建立皮肤疾病的生物标志物并降低临床研究参与者的风险。与传统皮肤活检技术相比，吸收性微生物活检操作可在数秒内完成，且由于其设计简单，无需专门培训。在本报告中，我们描述了吸收性微生物活检在一名志愿者身上的使用方法，包括加载和应用。然后，我们展示了如何从吸收的样本中分离RNA。最后，我们演示了如何使用定量逆转录PCR（RT-qPCR）来量化血液（CD3E和CD19）和皮肤（KRT14和TYR）的mRNA表达水平。我们所描述的方法使用的是现成的试剂盒和试剂。该方案提供了一种微创方法，可在同一吸收性微生物活检基质中同时对皮肤和血液进行采样。我们发现人类伦理委员会、临床医生和志愿者都支持这种皮肤病学研究方法。
