Journal
MOLECULAR BIOTECHNOLOGY
Volume 50, Issue 1, Pages 49-56Publisher
HUMANA PRESS INC
DOI: 10.1007/s12033-011-9414-6
Keywords
Hepatitis B virus; Circulating microRNA; Quantitative real-time PCR; Reference genes; Normalization
Funding
- National Key Projects for Infectious Disease of China [2008ZX10002-021]
- Program of Shanghai Chief Scientist [08XD1400800]
- National Natural Science Foundation of China [30700991]
- Fudan University
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Circulating microRNAs (miRNAs) were found to exist in serum/plasma in a highly stable, cell-free form, and aberrantly expressed in many human diseases. Currently, the expression levels of circulating miRNAs are estimated by quantitative real-time polymerase chain reaction. However, no study has systematically evaluated reference genes for evaluating circulating microRNA expression. This study describes the identification and characterization of an appropriate reference gene for the normalization of circulating miRNA levels in hepatitis B virus (HBV)-infected patients and healthy people. Ten miRNAs that resemble the mean expression of the TaqMan low density array together with U6, RNU6B, and miR-16 were validated with two algorithms, geNorm, and NormFinder, after ensuring their equivalent expression between the two study groups. The combination of miR-26a, miR-221, and miR-22* is recommended as the most stable set of reference genes for circulating miRNA evaluation in HBV patients and healthy people.
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