Establishment of a composition for early diagnosis of ovarian cancer
An ovarian cancer and individual technology, which is applied in the determination/inspection of microorganisms, biochemical equipment and methods, instruments, etc., can solve the problems of low tumor burden, low abundance of ctDNA mutations, no mutation detection or missed detection, etc., to achieve Effects of Specificity and Sensitivity Improvement
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[0097] The capture antibody can be prepared by linking a monoclonal antibody against the target biomarker to magnetic microparticles in the presence of a coupling solution under appropriate reaction conditions. Further, the detection antibody may be an alkaline phosphatase-labeled monoclonal antibody, an alkaline phosphatase-labeled monoclonal antibody, and combinations thereof.
[0098] As an embodiment of the protein chip of the present invention, the capture antibody is immobilized on microspheres carrying specific detectable signals (molecules) to prepare a liquid-phase protein chip. The principle of detection by using the liquid-phase protein chip is as follows: A single microsphere is passed through the detection channel, and the microsphere identification signal and detectable signal on the microsphere are simultaneously detected using two lasers. One type of laser excitation is the identification signal of the microspheres on the microspheres. According to the differen...
Embodiment 1
[0153] 1. Blood Sample Collection
[0154] Venous blood was collected using cell-free DNA blood collection tubes (LBgard blood collection tubes, Biomatirca, USA). blood cell, WBC) separation. All subjects underwent 10 mL whole blood sample collection. Plasma and peripheral blood samples were drawn before anesthesia on the day of surgery. The plasma of healthy volunteers in the biobank is stored at -80°C, no less than 3mL, and the storage time is within 2 years. It is transported to Shanghai Yunsheng Medical Laboratory by dry ice.
[0155] 2. Extraction of cfDNA and genomic DNA
[0156] Peripheral blood (10 mL for each newly collected sample, 3 mL for biobank samples) was centrifuged at 4° C., 1600 g for 10 minutes. The plasma (upper yellow layer) and leukocytes (middle layer) were collected, and then the upper plasma was centrifuged a second time to remove residual cells or cell debris at 4°C, 16000 g, 10 minutes. 0.5 mL of plasma was isolated and stored separately for su...
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