Molecular marker for diagnosing primary biliary cholangitis and application thereof
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A molecular marker, cholangitis technology, applied in the field of molecular biology, can solve the problems of PBC dependence and misdiagnosis, and achieve the effect of reducing missed diagnosis and misdiagnosis, simple sampling and less trauma
Pending Publication Date: 2022-02-25
THE FIRST AFFILIATED HOSPITAL OF ARMY MEDICAL UNIV +1
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Many studies in recent years have shown that the high expression of human multimeric immunoglobulin receptors in the liver is a risk factor for liver fibrosis, which may be related to hepatitis virus infection and hepatic stellate cell transdifferentiation. Human multimeric immunoglobulin receptors can also Promote the metastasis of human hepatocellular carcinoma (Hepatocellular carcinoma, HCC), but whether human multimeric immunoglobulin receptor is involved in the occurrence and development of PBC and the value of anti-human multimeric immunoglobulin receptor antibody in the diagnosis of PBC have not been reported yet , this indicator has not been applied clinically
[0004] Serum AMAs and their M2 subtypes are currently important auxiliary diagnostic indicators for PBC in clinical practice. However, there is a certain rate of misdiagnosis and missed diagnosis of this indicator, so the diagnosis of PBC still mainly relies on liver biopsy
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Embodiment 1
[0035] Example 1: To clarify the expression and localization of human multimeric immunoglobulin receptors in the liver tissues of CTR, PBC, OC, SSC and NASH patients.
[0036] One case of liver tissue samples remaining after surgical resection or clinicopathological diagnosis of patients with CTR, PBC, OC, SSC and NASH in the inpatient department of Southwest Hospital was collected. The liver tissue samples were fixed with paraformaldehyde, dehydrated and transparent, embedded in wax and then sliced. The thickness of the sliced tissue was 4um. Multiple immunohistochemical fluorescence technology was used to detect the expression and localization of human multimeric immunoglobulin receptors in the liver tissue of the subject. The reagent used was a multiple immunohistochemical fluorescence kit (Absin, abs50014). The specific steps were as follows:
[0037] (1) Dewaxing to water: Paraffin slices were baked in a 65-degree oven for 1 hour; slices were soaked in fresh xylene for ...
Embodiment 2
[0052] Example 2: Determining the expression of serum anti-human multimeric immunoglobulin receptor antibodies in patients with CTR, PBC, and OC.
[0053] Peripheral blood samples from 12 CTR, 17 PBC, and 15 OC patients were collected from the inpatient department of Southwest Hospital, and the expression of anti-human multimeric immunoglobulin receptor antibody in the serum of the subjects was detected by enzyme-linked immunosorbent assay (ELISA) , the reagent used is human multimer immunoglobulin receptor antibody ELISA kit (Lun Changshuo Bio, 100712), and the specific steps are:
[0054] (1) Take serum:
[0055] Place the whole blood sample collected in the serum separation tube at room temperature for 2 hours or overnight at 4°C, then centrifuge at 1000×g for 20 minutes, then take the supernatant, or store the supernatant at -20°C or -80°C to avoid Freeze and thaw repeatedly.
[0056] (2) Preparation of reagents and equipment:
[0057] Microwell microtiter plate, standa...
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Abstract
The invention discloses a molecular marker for diagnosing primary biliary cholangitis and application thereof. According to the present invention, the anti-human multimer immunoglobulin receptor antibody is adopted as the primary biliary cholangitis diagnosis molecular marker so as to prepare the diagnosis reagent; the human multimer immunoglobulin receptor antibody is remarkably increased in serum of PBC AMA-M2 subtype positive and negative patients, which indicates that in the attack of PBC, the serum anti-human multimer immunoglobulin receptor antibody specifically targets human multimer immunoglobulin receptors on small and medium bile ducts in the liver, thereby causing damage to the small and medium bile ducts. According to the invention, a new explanation is provided for the pathogenesis of the PBC, and in addition, a new means is provided for the specific diagnosis of the PBC by detecting the level of the serum anti-human multimeric immune globulin receptor antibody. The method is simple and reliable, materials are convenient to obtain, and the method has great application value in PBC diagnosis.
Description
technical field [0001] The invention belongs to the technical field of molecular biology, and relates to a serum anti-human multimeric immunoglobulin receptor antibody, specifically, a serum anti-human multimeric immunoglobulin receptor antibody used for the diagnosis of primary biliary cholangitis Molecular markers and their applications. Background technique [0002] Primary biliary cholangitis (PBC) is a chronic cholestatic liver disease characterized by autoimmune destruction of small and medium-sized intrahepatic bile ducts, leading to progressive cholestasis, which can slowly develop into cirrhosis and liver disease. Exhausted. At present, there are more than 100,000 newly diagnosed cases of this disease in the world every year, which mainly occurs in women, and the prevalence rate may be as high as 0.1% in women over 40 years old. Previous studies have shown that genetic and environmental factors, immune disorders and other factors are involved in the pathogenesis o...
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