S-poly(T)plus荧光定量PCR技术在检测肝癌相关miR-199中的临床应用价值

吴娟; 张雅琴; 吴云彬; 陈敏; 于洲; 曹颖平

doi:10.3971/j.issn.1000-8578.2020.19.1618

S-poly(T)plus荧光定量PCR技术在检测肝癌相关miR-199中的临床应用价值

Clinical Value of S-poly (T) plus Real-time PCR in Detection of Liver Cancer-related miR-199

摘要

摘要:
目的探讨S-poly(T) plus荧光定量PCR法与普通Poly(A)加尾法检测外周血血清miR-199敏感度的优劣性。
方法采用荧光定量PCR S-poly(T) plus法和Poly(A)加尾法检测肝癌患者和健康人血清miR-199，分别从敏感度、线性关系和重复性三方面进行评估其潜在临床应用价值。
结果 S-poly (T) plus法检测健康人miR-199的Ct值显著低于Poly(A)加尾法(30.32±0.72 vs. 35.42±0.67, P < 0.0001);而且S-poly(T) plus法具有更好的线性关系(R₂=0.9843±0.0047 vs. R₂=0.9573±0.0045, P=0.0027)和更好的的重复性(CV=2.59% vs. CV=3.52%, P=0.010)。S-poly (T) plus法在检测肝癌患者外周血miR-199时具有更好的敏感度。
结论 S-poly (T) plus荧光定量PCR检测肝癌血清miR-199比Poly(A)加尾法敏感度更高、重复性及稳定性更优。

Abstract:
Objective To explore the advantages and disadvantages of S-poly(T) plus qRT-PCR and Poly (A) qRT-PCR methods in detecting peripheral blood serum miR-199.
Methods S-poly(T) plus RT-PCR and Poly(A) qRT-PCR methods were used to detect the serum miR-199 in patients with liver cancer and healthy controls in three aspects of sensitivity, linearity and repeatability to evaluate their potential clinical value.
Results The Ct values of S-poly(T) plus qRT-PCR method for miR-199 in healthy controls was significantly lower than that of Poly(A) qRT-PCR method (P < 0.0001); and the S-poly(T) plus qRT-PCR method had better linear correlation (P=0.0027) and repeatability (P=0.010). S-poly(T) plus qRT-PCR method had better sensitivity in detecting serum miR-199 in liver cancer patients comparing with Poly (A) qRT-PCR method.
Conclusion In the detection of liver cancer serum miR-199, S-poly(T) plus qRT-PCR method is more sensitive, repeatable and stable than Poly(A) qRT-PCR.

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