CAJ | 학술논문

【目的】建立人IFN‐λ1‐3型mRNA实时荧光定量聚合酶链式反应（qRT‐PCR）检测方法。【方法】根据人IFN‐λ1‐3型mRNA的基因序列合成特异性引物及探针，用体外转录方法制备IFN‐λ1‐3型的mRNA标准品，建立人IFN‐λ1‐3型mRNA qRT‐PCR检测方法。【结果】IFN‐λ1‐3型RNA标准品与对应的CT 值有良好的线性关系；扩增效率均在90％～100％之间；灵敏度分别是：1×100 co pies／μL、1×101 co pies／μL、1×101 co pies／μL ；变异系数均小于2％；各型间交叉反应不明显。【结论】本研究建立的IFN‐λ1‐3型mRNA qRT‐PCR检测方法灵敏度高，具有良好的重复性及特异性。
【목적】건립인IFN‐λ1‐3형mRNA실시형광정량취합매련식반응（qRT‐PCR）검측방법。【방법】근거인IFN‐λ1‐3형mRNA적기인서렬합성특이성인물급탐침，용체외전록방법제비IFN‐λ1‐3형적mRNA표준품，건립인IFN‐λ1‐3형mRNA qRT‐PCR검측방법。【결과】IFN‐λ1‐3형RNA표준품여대응적CT 치유량호적선성관계；확증효솔균재90％～100％지간；령민도분별시：1×100 co pies／μL、1×101 co pies／μL、1×101 co pies／μL ；변이계수균소우2％；각형간교차반응불명현。【결론】본연구건립적IFN‐λ1‐3형mRNA qRT‐PCR검측방법령민도고，구유량호적중복성급특이성。
[Objective] To develop a real‐time fluorescent quantitative polymerase chain reaction (PCR) for de‐tecting human interferon lambda1‐3 mRNA (IFN‐λ1‐3 mRNA) .[Methods] According to the sequences of human interferon lambda 1‐3 mRNA ,specific primers and fluorescent probes were designed .The standard products for real‐time fluorescent quantitative PCR were prepared by in vitro transcription .[Results] The quantitative assay of human interferon lambda1‐3 mRNA worked well with an excellent linearity .Amplification efficiency was between 90% ~100% while the sensitivities for IFN‐λ1‐3 mRNA genes were 100 ,101 and 101 copies/μl .All variation coef‐ficients of IFN‐λ1‐3 mRNA gene were lower than 2% .And cross reaction between each type of interferon lambda 1‐3 mRNA was not obvious .[Conclusion] The real‐time fluorescent quantitative PCR assay for IFN‐λ1‐3 mRNA gene is rapid ,sensitive ,specific and highly reproducible .Thus it provides rationales for further functional studies of IFN‐λ1‐3 mRNA gene .