Fluorescence quantitative kit for antiviral gene Mxa and detecting method thereof

Abstract

The invention relates to a reagent box, in particular to an anti-virus gene MxA fluorescence quantitative reagent box and the detection method thereof. The reagent box comprises MxA, housekeeping gene GAPDH amplification primers, DNA polymerase a probe used for PCR reaction, and one or more kind(s) of buffer solution thereof. The detection method comprises the following steps: firstly, peripheral blood mononuclear cell PBMC is gathered; secondly, PBMC cells with interferon are irritated; thirdly, a TA clone including MxA and housekeeping gene GAPDH target gene fragments is established; fourthly, real-time fluorescence PCR is adopted to detect the inducible expression of MxA mRNA; fiftyly, the Ct value of samples is judged according to the standard curve and the domain value. The anti-virus gene MxA fluorescence quantitative reagent box and the detection method of the invention have the advantages that after adopting the irritation of the peripheral blood interferon, the expression of the MxA can reflect the therapeutic effect of patients with interferon after being treated, thus patients and doctors have vital significance to select the interferon treatment, and the waste of medical resources is avoided.

Description

technical field [0001] The invention relates to a kit, in particular the invention relates to an antiviral gene MxA fluorescence quantitative kit and detection method. Background technique [0002] α-interferon (IFN-α) is currently one of the internationally recognized effective drugs for the treatment of chronic hepatitis B. After 6-12 months of IFN-α treatment in patients with chronic hepatitis B, the ALT recovery rate is 34% to 45%. The HBeAg disappearance / anti-HBe appearance rate is about 15% to 37%, the HBV-DNA negative conversion rate (less than 105 copies / ml) is 37%, and the HBsAg negative conversion rate is about 1%-8%. Clinicians are very concerned about the reasons why chronic hepatitis B patients achieve sustained response, partial response or no response after interferon treatment. Factors closely related to interferon response include high ALT level before treatment, low HBV-DNA level before treatment load and liver histologically evident inflammatory activity....

AI Technical Summary

Problems solved by technology

[0004] At present, the prediction and judgment of the curative effect of interferon mainly comes from the consideration of various clinical indicators of patients by clinicians, such as gender, age, ALT, HBV-DNA, HBeAg, etc., but these judgments lack accuracy and objectivity, and cannot improve the treatment effect. patient effectiveness

Due to the lack of public literature, the SNP predictive effect of interferon launched by Shanghai Jikang Co., Ltd. cannot be objectively evaluated.

However, the common problem of using SNP to predict the efficacy is that the complexity of the population and the SNP distribution have only three genotypes, and the distribution frequency may affect its value in the population as a whole. For example, MxA-88G / T SNP is related to the efficacy of interferon , but the frequency of the T allele that tends to be effective in the Chinese population is only 10%, which excludes the possibility of treatment for most potentially effective patients

Images