Application of phosphoglycerate kinase 1 to preparation of rheumatoid arthritis diagnosis reagent
A diagnostic reagent, arthritis technology, applied in the field of application of phosphoglycerate kinase 1 in the preparation of rheumatoid arthritis diagnostic reagents, to achieve the effect of high specificity and strong sensitivity
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Embodiment 1
[0064] Example 1: Discovery and verification of PGK1 as a new blood marker for clinical diagnosis of rheumatoid arthritis
[0065] 1.1 Establishment of type II collagen-induced arthritis (CIA) rat model:
[0066] Male Wistar rats (n=20, each rat weighing 150-200 g) were obtained from the Animal Science and Health Science Experimental Center of Peking University (Beijing, China). These mice were randomly divided into two groups, CIA group and control group. Mice in all experimental groups were intradermally injected with a CFA solution containing 500 μg of bovine collagen II (SIGMA-ALDRICH, USA) at the base of their tails. And after 14 days, use the same method to inject again. In the same way, the control group was injected with BSA instead of type II collagen at the base of the tail. CIA rats were clinically scored according to the clinical manifestations of each carpal tarsal joint, and the evaluation criteria were as follows: 0=normal, 1=slight edema and erythema, 2=obvi...
Embodiment 2
[0096] Example 2: Diagnostic reagents and detection methods for clinical diagnosis of rheumatoid arthritis
[0097] Using PGK1 as a blood marker of rheumatoid arthritis, a diagnostic reagent for clinical diagnosis of rheumatoid arthritis is prepared, and blood of patients is detected. Blood samples were obtained by standard venipuncture from RA patients (n=120, 75 females, aged 20–75 years). Chronic joint inflammation for at least 3 years, erythrocyte sedimentation rate (ESR, 19 to 22 mm / hour, average 44 mm / hour), c-reactive protein (8-196 U / ml, average 29.1 U / ml) and rheumatoid factor (10.5- 2090U / ml) levels increased. Combined therapy with methotrexate and leflunomide for at least two months. 120 (80 females; 24-58 years old, average 40.2 years old) healthy blood donors. Blood samples were placed in EDTA-K2 vacuum blood collection tubes.
[0098] The composition of the diagnostic reagent includes: carbonate buffer solution, PBST washing solution, blocking solution, chrom...
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