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Methodology and Validation
a Department of Chemical Pathology, The Chinese University of Hong Kong, Prince of Wales Hospital, Shatin, New Territories, Hong Kong SAR, China
Key Words: plasma RNA • integrity • quantitative analysis
Address for correspondence: Prof. Y. M. Dennis Lo, Department of Chemical Pathology, The Chinese University of Hong Kong, Prince of Wales Hospital, Rm 38023, 1/F Clinical Sciences Building, 30-32 Ngan Shing Street, Shatin, New Territories, Hong Kong SAR, China. Voice: +852-2632-2563; fax: + 852-2636-5090. e-mail: loym{at}cuhk.edu.hk; blend{at}cunk.edu.hk.
The detection of cell-free RNA in plasma and serum of human subjects has found increasing applications in the field of medical diagnostics. However, many questions regarding the biology of circulating RNA remain to be addressed. One issue concerns the molecular nature of these circulating RNA species. We have recently developed a simple and quantitative method to investigate the integrity of plasma RNA. Our results have suggested that cell-free RNA in plasma is generally present as fragmented molecules instead of intact transcripts, with a predominance of 5' fragments. In this article, we summarize the basic principles in the experimental design for plasma RNA integrity analysis and highlight some of the important technical considerations for this type of investigation.
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