title: Identification of suitable reference genes for qRT-PCR analysis of circulating microRNAs in hepatitis B virus-infected patients.
abstract: 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.
authors.affiliation: Live Cancer Institute and Zhongshan Hospital, Institutes of Biomedical Sciences, Fudan University, Shanghai 200032, China. zhuht02@yahoo.com.cn
authors.forename: Hai-Tao
authors.initials: HT
authors.lastname: Zhu
authors.forename: Qiong-Zhu
authors.initials: QZ
authors.lastname: Dong
authors.forename: Guan
authors.initials: G
authors.lastname: Wang
authors.forename: Hai-Jun
authors.initials: HJ
authors.lastname: Zhou
authors.forename: Ning
authors.initials: N
authors.lastname: Ren
authors.forename: Hu-Liang
authors.initials: HL
authors.lastname: Jia
authors.forename: Qing-Hai
authors.initials: QH
authors.lastname: Ye
authors.forename: Lun-Xiu
authors.initials: LX
authors.lastname: Qin
mesh_terms: D000328:Adult; D005260:Female; D020869:Gene Expression Profiling / Q000379:methods / Q000592:standards*; D006515:Hepatitis B virus; D019694:Hepatitis B, Chronic / Q000235:genetics* / Q000473:pathology / Q000821:virology; D006801:Humans; D008297:Male; D035683:MicroRNAs / Q000097:blood* / Q000235:genetics; D008875:Middle Aged; D020411:Oligonucleotide Array Sequence Analysis; D012015:Reference Standards; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000592:standards*
chemical_list: D035683:MicroRNAs; C548375:MIRN22 microRNA, human; C525709:MIR221, human; C528163:MIRN26 microRNA, human