Gene diagnosis reagent kit for systemic lupus erythematosus

A lupus erythematosus and genetic diagnosis technology, applied in the field of molecular biology, can solve the problems that diagnosis plays a big role, time-consuming and labor-intensive, and increases consumption

Inactive Publication Date: 2016-03-23
THE AFFILIATED DRUM TOWER HOSPITAL MEDICAL SCHOOL OF NANJING UNIV
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  • Abstract
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  • Claims
  • Application Information

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Problems solved by technology

Later, I tried to detect the expression of 5 interferon-induced genes, and found that it has certain

Method used

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  • Gene diagnosis reagent kit for systemic lupus erythematosus
  • Gene diagnosis reagent kit for systemic lupus erythematosus
  • Gene diagnosis reagent kit for systemic lupus erythematosus

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] 1. Gene identification and primer optimization

[0030] According to the previous research, the type I interferon-induced genes related to the diagnosis of SLE were determined to be LY6E, ISG15 and OASL. Through a large number of repeated experiments, combined with the optimization of the template amount and the Tm value of the primers, the primers for amplifying the above three genes were determined. and primers to amplify the housekeeping gene RPLPO.

[0031] For specific primer sequences, see the sequence listing (OASL: SEQ ID NO.1-2; ISG15: SEQ ID NO.3-4; LY6E: SEQ ID NO.5-6; RPLPO: SEQ ID NO.7-8).

[0032] 2. Processing of samples to be tested

[0033] In the morning, 2-3ml of venous whole blood was collected with EDTA venous vacuum disposable blood collection tubes (BD Biosciences, USA) in a resting state on an empty stomach, and then RNA was extracted with Trizol (Invitrogen, USA), and reversed with SuperscriptII reverse transcription kit (Invitrogen, USA). rec...

Embodiment 2

[0041] 1. Comparing the changes of modified interferon index between SLE patients and normal population and patients with other autoimmune diseases

[0042]Blood samples were collected from normal people, SLE patients and patients with other autoimmune diseases, the relative expression values ​​of each gene were obtained according to Example 1, and the expression levels of LY6E, ISG15 and OASL genes in each group were compared. Standardize the level of each gene by (relative gene expression value of the sample to be tested-average relative gene expression value of the normal group) / standard deviation of gene expression in the normal group, and then add the three values ​​corresponding to LY6E, ISG15 and OASL to obtain The modified interferon index was used to compare the differences in the levels of the modified interferon index in each group. Compared with normal people and patients with other autoimmune diseases, the expression levels of LY6E, ISG15, OASL and modified interf...

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Abstract

The invention relates to a gene diagnosis reagent kit for systemic lupus erythematosus. The gene diagnosis reagent kit comprises primers for amplification of the OASL gene, the IGS15 gene and the LY6E gene, and further comprises a primer for the house-keeping gene RPLPO and an amplification reagent. The gene diagnosis reagent kit overcomes the defects that an I type interferon induction gene lacks of specificity, the number of detection genes is large, much time and labor are consumed, and consumption is increased. A real-time fluorescence quantification PCR method is adopted, and the gene diagnosis reagent kit has the advantages of being fast, sensitive, accurate and the like; the expression of the LY6E gene, the ISG15 gene and the OASL gene is detected through a real-time fluorescence quantification PCR, improved interferon indexes are worked out, early diagnosis of an SLE patient is facilitated, an important basis and reference value can be provided for clinical disease confirmation, and thus early prevention and treatment of the disease are facilitated; the gene diagnosis reagent kit has application and popularization value.

Description

technical field [0001] The invention relates to a gene diagnosis kit for autoimmune diseases, more specifically a gene diagnosis kit for systemic lupus erythematosus, belonging to the technical field of molecular biology. Background technique [0002] Autoimmune venereal disease is a neglected, common chronic disease that causes death and disability. Systemic lupus erythematosus (systemic lupus erythematosus, SLE) is considered to be a prototype autoimmune disease, which can affect any organ of the body. The diagnosis of SLE is currently mainly based on the classification criteria formulated by the American College of Rheumatology in 1982 and revised in 1997, and updated by the Systemic Lupus Erythematosus International Collaborative Group (SLICC) in 2009. However, these SLE classification criteria all aim to separate SLE from the spectrum of autoimmune diseases, which is not conducive to the identification or detection of mild SLE and early SLE. For SLE, delay in treatmen...

Claims

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Application Information

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IPC IPC(8): C12Q1/68
Inventor 冯学兵孙凌云
Owner THE AFFILIATED DRUM TOWER HOSPITAL MEDICAL SCHOOL OF NANJING UNIV
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