A real-time fluorescent quantitative PCR kit for quantitative detection of mRNA content in the body, detection method and application

Abstract

The invention provides a real-time fluorescence quantitative PCR kit for quantitatively detecting mRNA content in a body, a detection method and an application, and relates to the technical field of mRNA detection. The kit of the present invention is based on RT-PCR, uses mRNA as a standard to construct a standard curve, and the diluted serial standard solution passes through the same reverse transcription and qPCR process as the sample to be tested, so as to better simulate the sample detection process , visually display the content of mRNA drugs, reduce the construction work of plasmids, and realize the rapid and accurate quantitative detection of mRNA drugs in biological samples. It has the advantages of simple operation, high sensitivity, wide detection range, and strong specificity, and can be used for mRNA drugs Preclinical drug biodistribution studies, clinical research biological sample testing, and drug distribution monitoring in clinically treated patients.

Description

technical field [0001] The invention belongs to the technical field of mRNA detection, and in particular relates to a real-time fluorescence quantitative PCR kit for quantitative detection of mRNA content in a body, a detection method and an application. Background technique [0002] mRNA drugs are promising molecular therapeutics in recent years, and have great application potential in the prevention of infectious diseases, metabolic diseases, cancer and the treatment of various diseases including rare diseases. The principle of mRNA drugs is based on the characteristics of mRNA guiding protein synthesis. After necessary chemical modification, mRNA enters the cytoplasm through different delivery systems, and uses its own nucleotides in the cytoplasm for transcription and expression to generate the proteins needed by the body. Because mRNA drugs do not enter the nucleus, do not change the genome, have transient activity, and can be degraded through physiological metabolism, ...

AI Technical Summary

Problems solved by technology

However, so far, no mRNA drug has been approved for marketing by the US FDA, and most mRNA drugs are still in the clinical research stage. In the process of research and development of mRNA drugs, it is still necessary to establish a systematic, complete, and stable technical solution for detection, analysis, and verification. , the technology used requires the ability to specifically identify and detect the sequence of mRNA drugs in biological samples from different sources, so far no commercial kits have been realized

Traditional mRNA quantitative detection methods such as Northern blot and in situ hybridization are not suitable for the quantitative detection of mRNA drugs due to the disadvantages of complicated operation, possibility of radioactive contamination, long time consumption, and low sensitivity.

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