CN-116334208-B - Internal reference gene suitable for miRNA detection, related products and application thereof

Abstract

The invention discloses an internal reference gene suitable for miRNA detection, and a related product and application thereof, and relates to the field of biological detection. According to the invention, experimental tests and data analysis of different methods are carried out on miRNAs in a heating control group and a KD infant group, and the micro ribonucleic acid miR-30d-5p is found to be used as an internal reference gene for detecting and standardizing blood platelet microRNA of Kawasaki disease, and has good stability and accuracy.

Inventors

• LIU JIA
• LI SHIMING
• XIE LIJIAN
• HUI WEI
• Xue Shuaixiang
• LI GUANG
• HUANG JIANHANG

Assignees

• 道之精准医学科技(上海)有限公司

Dates

Publication Date

20260505

Application Date

20230117

Claims (8)

1. 1. The application of the miR-30d-5p detection reagent in preparation of a detection reagent of an internal reference gene suitable for detection of miRNA, wherein the miRNA is a miRNA marker for detecting Kawasaki disease.
2. 2. The use of claim 1, wherein the detection reagent for miR-30d-5p comprises at least one of a primer pair, a probe, and a chip for detecting miR-30d-5 p.
3. 3. Application of miR-30d-5p detection reagent in preparation of product for detecting Kawasaki disease.
4. 4. The method of claim 3, wherein the product is selected from the group consisting of reagents, kits, chips and predictive models.
5. 5. The use of claim 3 or 4, wherein the detection reagent for miR-30d-5p comprises at least one of a primer pair, a probe, and a chip for detecting miR-30d-5 p.
6. 6. A reagent or a kit is characterized by comprising a detection reagent of miRNA and a detection reagent of an internal reference gene, wherein the internal reference gene comprises miR-30d-5p; the miRNA detection reagent is used for detecting miRNA markers of Kawasaki disease.
7. 7. The reagent or kit according to claim 6, wherein the miRNA markers for detecting Kawasaki disease comprise let-7g-5p and miR-26a-5p.
8. 8. The reagent or kit according to claim 6 or 7, wherein the detection reagent for the internal reference gene comprises at least one of a primer pair, a probe and a chip for detecting miR-30d-5 p.

Description

Internal reference gene suitable for miRNA detection, related products and application thereof Technical Field The invention relates to the field of biological detection, in particular to an internal reference gene suitable for miRNA detection, and a related product and application thereof. Background Kawasaki Disease (KD) is an acute, self-limiting vasculitis, mainly occurring in inflammatory syndromes in children. The disease mainly involves middle and small arteries, especially coronary arteries, and is the main cause of acquired diseases. Kawasaki disease may occasionally lead to death, especially when missed or not given timely treatment. KD is characterized by a long duration of fever, at least with 4-5 clinical features of more than five days fever, limb changes (hard swelling of hands and feet, ecdysis), rashes, non-exudative conjunctivitis, oral changes, and cervical lymphadenopathy (usually unilateral). Approximately one-fourth of KD patients have incomplete clinical manifestations, i.e. the patients have insufficient primary clinical manifestations and can delay diagnosis leading to higher risk of disease. Thus leading to incomplete clinical diagnostic criteria for kawasaki disease. Although studies are currently being made on the reliability of diagnosis, a variety of new biomarkers and classification tools, KD-specific biomarkers have not been found so far. In view of this, the present invention has been made. Disclosure of Invention The invention aims to provide an internal reference gene suitable for miRNA detection, and a related product and application thereof. The invention is realized in the following way: In a first aspect, the embodiment of the invention provides an application of a miR-30d-5p detection reagent in preparation of a detection reagent of an internal reference gene suitable for miRNA detection. In a second aspect, embodiments of the invention provide the use of a detection reagent for miR-30d-5p in the preparation of a product for detecting Kawasaki disease. In a third aspect, the embodiment of the invention provides a reagent or a kit, which comprises a detection reagent of miRNA and a detection reagent of an internal reference gene, wherein the internal reference gene comprises miR-30d-5p. In a fourth aspect, embodiments of the present invention provide a method of detecting a target miRNA expression level in a sample, the method comprising using miR-30d-5p as an internal reference gene for analyzing the target miRNA expression level. The invention has the following beneficial effects: according to the invention, through experimental tests and data analysis of different methods of miRNAs in a heating control group and a KD infant group, the micro ribonucleic acid miR-30d-5p can be used as an internal reference gene for detecting and standardizing blood platelet microRNA of Kawasaki disease, and has good stability and accuracy. Drawings In order to more clearly illustrate the technical solutions of the embodiments of the present invention, the drawings that are needed in the embodiments will be briefly described below, it being understood that the following drawings only illustrate some embodiments of the present invention and therefore should not be considered as limiting the scope, and other related drawings may be obtained according to these drawings without inventive effort for a person skilled in the art. FIG. 1 is a comparison of average expressed Ct values for 4 miRNAs; FIG. 2 is a comparison of the coefficients of variation of 4 miRNAs; FIG. 3 shows the stability of gene expression of 4 miRNA in the geNorm analysis; FIG. 4 shows the stability values of gene expression of 4 mRNAs analyzed by NormFinder; FIG. 5 shows miR-126-3p normalized let-7g-5p relative expression (A) and miR-30d-5p normalized let-7g-5p relative expression (B); FIG. 6 is a plot of miR-26a-5p relative expression normalized to miR-126-3p (A) and miR-26a-5p relative expression normalized to miR-30d-5p (B); FIG. 7 is a ROC graph of miR-126-3p and miR-30d-5p in which let-7g-5p is normalized, respectively; FIG. 8 is a ROC graph of miR-126-3p and miR-30d-5p normalized miR-26a-5p, respectively. Detailed Description In order to make the objects, technical solutions and advantages of the embodiments of the present invention more clear, the technical solutions of the embodiments of the present invention will be clearly and completely described below. The specific conditions are not noted in the examples and are carried out according to conventional conditions or conditions recommended by the manufacturer. The reagents or apparatus used were conventional products commercially available without the manufacturer's attention. MiRNA (microRNA) endogenous non-coding small molecule RNA for regulating gene expression at posttranscriptional level, and the length is about 22 nucleotides. More and more studies have shown that mirnas play an important role in the regulatory mechanisms of various organisms, including de