Esophagus cancer early combined detection kit
A combined detection and kit technology, applied in the field of biomedicine, can solve the problems of limited promotion of endoscopic screening, trauma and high cost of endoscopic screening, and achieve good clinical application prospects, less trauma and less side effects
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Embodiment 1
[0022] Example 1: The detection of eight kinds of serum autoantibodies on the early diagnosis test of esophageal cancer
[0023] 1. Research object
[0024] From March 2015 to February 2018, 300 serum samples were collected from the First Affiliated Hospital of Zhengzhou University, including 100 patients with early EC, 100 patients with EBL, and 100 patients with HC. All 100 patients with early EC were diagnosed based on endoscopy combined with cytology or histopathology. They did not receive any anti-tumor treatment such as chemotherapy and radiotherapy. The TNM classification was stage 0, I, and II. 100 cases of EBL patients were from patients who were initially treated in the First Affiliated Hospital of Zhengzhou University during the same period, including benign esophageal tumors, benign esophageal ulcers, esophageal erosions, and reflux esophagitis. The 100 HC patients were healthy adults from the Physical Examination Center of the First Affiliated Hospital of Zhengzh...
Embodiment 2
[0035]Example 2 Research on the Early Diagnosis Test of Esophageal Cancer by Detection of Four Kinds of Serum Autoantibodies
[0036] Based on the four autoantibody markers screened out through the small sample test in Example 1, in order to determine that these four autoantibody markers have a good early diagnostic value for EC, the present invention tested 1138 cases of serum through the ELISA method. In-depth research on the value of anti-C-myc antibody, anti-HER-2 antibody, anti-Cyfra21-1 antibody and anti-P53 antibody detection in the early diagnosis of EC in a large sample range, the research cohort includes 818 cases of training set and 320 cases of independent matching validation set.
[0037] The model building process for the joint detection of antigens corresponding to the four autoantibody markers mainly includes two stages: the first stage, through 818 samples of the training set, including 258 early EC patients, 260 EBL patients and 300 HC patients, four ELISA d...
Embodiment 3
[0050] Embodiment 3: the composition of kit
[0051] The composition of kit of the present invention is as follows:
[0052] (1) Antigen-coated 96-well enzyme-labeled reaction plate
[0053] Its preparation method is as follows:
[0054] .Luminex microspheres coupled with antigenic protein, the specific process is:
[0055] Transfer 5.0×106 raw Luminex microspheres to a microcentrifuge tube; and add 10 µL 150 mg / mL sulfo-NHS and 10 µL 150 mg / mL EDC to the centrifuge tube, incubate at room temperature for 20 minutes; then use 50 mM pH 5.0 Wash the activated Luminex microspheres with MES to remove excess sulfogroup-NHS and EDC; add 10 µg of antigenic protein to a microcentrifuge tube, then add 50 mM MES with pH 5.0 to make the total volume to 500 µL, and incubate at room temperature for 2 hours; coupled Luminex microspheres were washed twice with PBS containing 0.1% (v / v) BSA, 0.02% (v / v) Tween-20, 0.05% (v / v) sodium azide pH 7.4 Afterwards, store at 2-8°C in the dark for ...
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