The Specificity, Sensitivity and Clinical Correlation of CBA, CBA-TSA, RIPA and ELISA Assay in Detecting AChR and MuSK IgG
Study Details
Study Description
Brief Summary
Myasthenia gravis (MG) is a neuromuscular junction (NMJ) transmission disorder mediated by autoantibodies against AChR, MuSK and other autoantigens located at the post synaptic membrane of neuromuscular junction. Presence of autoantibody specific for AChR or MuSK can establish diagnosis in conjunction with clinical presentations. In the guidelines for the diagnosis and treatment of myasthenia gravis in the United States or the United Kingdom, determination of AChR and MuSK antibodies has been recommended. Radioimmunoprecipitation assay (RIPA), enzyme-linked immunosorbent assay (ELISA) and cell-based assay (CBA) that are all commercially available, have been adopted for detection by most referring neurologists. At present, specificity and sensitivity of these methods have not been compared in large cohorts in the context of stringent quality control. As a consequence, there are no national or international consensus regarding selection of methods and interpretation of results, resulting in challenges to neurologists managing these patients. To this end, we propose to conduct a multicenter, double-blind, prospective study to compare the sensitivity and specificity of CBA, CBA-TSA (Tyramide Signal Amplification), RIPA and ELISA assays.
| Condition or Disease | Intervention/Treatment | Phase |
|---|---|---|
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Detailed Description
Participants: The serum of myasthenia gravis patients.
Primary aim: Comparison of the specificity, sensitivity and clinical correlation of CBA, CBA-TSA, RIPA and ELISA assay in detecting AChR and MuSK IgG of myasthenia gravis. Secondary aim: Analyzing the advantage of CBA-TSA assay when AChR or MuSK IgG in low abundance and low affinity. To verify the function of AChR subunits in detecting the AChR antibody IgG through CBA testing.
Study design: Multicenter, double-blind, CBA/CBA-TSA, ELSIA and RIPA methodology comparison
Total cases: 1) Recruiting 3000 patients with suspected myasthenia gravis. The Beijing-Tianjin Center has already recruited 1,500 cases. At least 50 cases for each other centers, competitive recruited.
- Recruiting 200 people as normal controls that has already completed by the Beijing-Tianjin Center.
Trial Period: The trial recruiting duration is 5-6 months, and the entire study period is estimated to be about 8 months.
Research reagents: 1. The method of CBA and CBA-TSA antibody IgG detected was performed by Tianjin Jinyu Medical Laboratory Co., Ltd. The kit of AChR or MuSK IgG CBA and CBA-TSA was purchased from Tianjin New Terrain Biological Technology Co., Ltd, China. 2. The method of RIPA was performed by Beijing North Biotechnology Co., Ltd. The kit of AChR or MuSK IgG RIPA was purchased by RSR Limited, UK. 3. The method of ELISA was performed by Tianjin Medical University General Hospital Neuroimmunology Laboratory. The kit of AChR or MuSK IgG ELISA Kits was purchased by RSR Limited, UK or IBL Limited, Germany.
Groups: The samples were divided into 4 groups according to the detected methods: CBA group, CBA-TSA group, RIPA group and ELISA group.
Study Steps: 1. After MG suspected patients in each center matched up with the inclusion and exclusion criteria, each patient's serum was divided into 4 equal parts (500 ul/part), and were randomly assigned to the CBA, ELSIA and RIPA testing laboratories for detecting. 2. Based on the standard process of kits, the laboratories of CBA, CBA-TSA, ELISA or RIPA completed the detection of the participants serum, then recorded the original value, control or reference value. Data analysis of sensitivity, specificity and clinical correlation of each method were summarized by Beijing-Tianjin Neuroimmunity/New Terrain. 3. The difference comparison between CBA and CBA-TSA was completed by Beijing-Tianjin Neuroimmunity Center. To verify the function of AChR subunits in detecting the AChR antibody IgG through CBA assay. 4. Each center will give the feedback of the final disease result of MG or non-MG, classification (GMG or OMG), clinical diagnosis and treatment data of the participants to the investigator. The investigator will complete the evaluation of respective clinical correlation of the CBA, ELSIA and RIPA methods. 5. The data statistics, analysis and summary of the project are supervised by a third-party audit company.
Evaluation indicators:
Prospective: 1. Technical indicators: 1) Evaluate the specificity, sensitivity and clinical correlation of the AChR and MuSK IgG detection through the CBA/CBA-TSA, ELISA and RIPA methods. 2) Determine the function of AChR subunits in detecting AChR antibody IgG through CBA. 3. Analysize the advantage of modified CBA method (CBA-TSA) in detecting the AChR or MuSK IgG in low abundance and low affinity. 4) Generate the first degree evidence in the testing methods of MG AChR and MuSK antibody detection.
Study Design
Arms and Interventions
| Arm | Intervention/Treatment |
|---|---|
| Cell Based Assay (CBA) Use AChR/MuSK IgG CBA Kit (Tianjin New Terrain Biological Technology Co., Ltd, China) to detect AChR and MuSK IgG of myasthenia gravis |
Diagnostic Test: CBA Assay
Comparsion of specificity, sensitivity and clinical correlation of CBA, CBA-TSA, ELISA and RIPA assay in autoantibodies detection of myasthenia gravis
Other Names:
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| CBA-TSA Assay Modified CBA method, Use AChR/MuSK CBA-TSA Kit (Tianjin New Terrain Biological Technology Co., Ltd, China) to detect AChR and MuSK IgG of myasthenia gravis |
Diagnostic Test: CBA Assay
Comparsion of specificity, sensitivity and clinical correlation of CBA, CBA-TSA, ELISA and RIPA assay in autoantibodies detection of myasthenia gravis
Other Names:
|
| RIPA Assay Use AChR/MuSK Radioimmunoassay Kit (RSR Limited, UK) to detect AChR and MuSK IgG of myasthenia gravis |
Diagnostic Test: CBA Assay
Comparsion of specificity, sensitivity and clinical correlation of CBA, CBA-TSA, ELISA and RIPA assay in autoantibodies detection of myasthenia gravis
Other Names:
|
| ELISA Assay Use AchR ELISA Kit (RSR Limited, UK) and MuSK ELISA Kit (IBL Limited, Germany) to detect AChR and MuSK IgG of myasthenia gravis |
Diagnostic Test: CBA Assay
Comparsion of specificity, sensitivity and clinical correlation of CBA, CBA-TSA, ELISA and RIPA assay in autoantibodies detection of myasthenia gravis
Other Names:
|
Outcome Measures
Primary Outcome Measures
- Comparsion of the specificity, sensitivity and clinical correlation [8 months]
Comparsion the specificity,sensitivity and clinical correlation of CBA, CBA-TSA, RIPA and ELISA assay in autoantibodies detection of myasthenia gravis
Secondary Outcome Measures
- To verify the differences of specificity and sensitivity in CBA and CBA-TSA assay [8 months]
Analyzing the advantage of CBA-TSA in detecting the AChR or MuSK IgG in low abundance and low affinity.
- To confirm the function of AChR subunits in detecting clustered AChR IgG [8 months]
To verify the binding function of AChR subunits in AChR antibody through the CBA assay.
Eligibility Criteria
Criteria
Inclusion Criteria:
- Suspected patients with myasthenia gravis present the clinical features of GMG(Generalized Myasthenia Gravis, GMG) or OMG(Ocular Myasthenia Gravis, OMG) need to be confirmed by serum antibody testing.
Exclusion Criteria:
- Abnormal serum samples, such as hemolysis or lipemia, which will affect the detection base value and the final interpretation of CBA, CBA-TSA, RIPA and ELISA; The samples with incomplete clinical data, which will affect the disease characterization.
Contacts and Locations
Locations
| Site | City | State | Country | Postal Code | |
|---|---|---|---|---|---|
| 1 | Tianjin Medical University General Hospital | Tianjin | China | 300000 |
Sponsors and Collaborators
- Tianjin Medical University General Hospital
Investigators
None specified.Study Documents (Full-Text)
None provided.More Information
Publications
None provided.- IRB2021-YX-260-01