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- W2386010772 abstract "Objective: To establish a SYBR GreenⅠreal-time fluorescent quantitative PCR(qRT-PCR) method for the detection of human RORγt(retinoid-related orphan receptor gammat) mRNA.Methods: The specific primers were designed according to the conserved sequence of human RORγt mRNA.Total RNA was extracted from human peripheral blood mononuclear cells(PBMC) stimulated by PMA and ionomycin,and was transcribed reversely into cDNA.The RORγt gene was obtained by PCR and cloned into PMD-18 vector by TA clone technology.The linearity,specificity and repeatability of the method were evaluated.The relative expression level of human RORγt mRNA in PBMCs from patients with GD was detected by this method. Results: The coefficient of correlation of the standard curve of this method was 0.998,melting curve was single peak,and the CVs in intra and inter-assay were 6.5%,8.4% and 9.6%,11.2% in high and low copies plasmid standards respectively.The relative expression level of human RORγt mRNA in GD group was higher markedly than that in healthy control group(t=-3.72,P0.01). Conclusion: qRT-PCR was a sensitive,specific and repeatable method for the detection of human RORγt mRNA,and this method serves as a technologic base for clinical application." @default.
- W2386010772 created "2016-06-24" @default.
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- W2386010772 date "2009-01-01" @default.
- W2386010772 modified "2023-09-28" @default.
- W2386010772 title "Establishment of the real-time fluorescent quantitative PCR for the detection of human RORγt mRNA" @default.
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