Detection of hepatitis C virus by single-step hairpin primer RT-PCR

被引:9
|
作者
Takei, Fumie [1 ]
Tani, Hideki [2 ]
Matsuura, Yoshiharu [2 ]
Nakatani, Kazuhiko [1 ]
机构
[1] Osaka Univ, Inst Sci & Ind Res, Ibaraki, Osaka 5670047, Japan
[2] Osaka Univ, Microbial Dis Res Inst, Suita, Osaka 5650871, Japan
关键词
Fluorescent molecule; PCR; Hairpin primer; HCV; Detection method; POLYMERASE-CHAIN-REACTION; REAL-TIME PCR; NUCLEIC-ACID HYBRIDIZATION; DYE CONCENTRATION; DNA; AMPLIFICATION; MICROBIOLOGY; TEMPERATURE; DIAGNOSIS; PRODUCTS;
D O I
10.1016/j.bmcl.2013.10.021
中图分类号
R914 [药物化学];
学科分类号
100701 ;
摘要
We developed a novel single-step virus detection system using the fluorescent molecule with a hairpin primer on the reverse transcription (RT)-polymerase chain reaction (PCR). Primers that are specific to Hepatitis C virus (HCV) having a hairpin tag at the 5 '-end were used for RT and PCR. The HCV-RNA template was reverse transcribed by reverse transcriptase with the hairpin primer (HP), and the resulting cDNA was amplified directly by HP-PCR (RT-HP-PCR). Using the RT-HP-PCR, we succeeded in demonstrating the detection of HCV-RNA in single-step. The RT-HP-PCR selectively detected HCV-RNA extracted from patients' sera containing contaminant materials. The method could be applicable to the quantitative detection of HCV. (c) 2013 Elsevier Ltd. All rights reserved.
引用
收藏
页码:394 / 396
页数:3
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