Tumor marker detection kit based on variable lymphocyte receptors and method for preparing same
A technology for tumor markers and detection kits, applied in the field of tumor marker detection kits and their preparation, to achieve the effects of low production cost, simple clinical detection, and convenient storage and transportation
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Embodiment 1
[0014] Example 1: Tumor marker ELISA detection kit
[0015] The capture reagent and detection reagent in the ELISA detection kit are replaced by the recombinant variable lymphocyte receptor (capture VLR and detection VLR) with high specificity and high affinity with tumor markers to replace the existing tumor marker ELISA detection kit The capture antibody, detection antibody, and other reagents are the same as the existing tumor marker ELISA detection kit.
[0016] The preparation method for capturing VLR and detecting VLR is:
[0017] a. Using the ovarian cancer tumor marker CA125 standard as an antigen, immunostimulate jawless vertebrates; take the blood of the immunized animals and separate lymphocytes; extract total RNA of lymphocytes, and use the universal primer Oligo-dT Reverse transcribe the mRNA into cDNA; design primers based on the conserved regions at both ends of the VLR gene and introduce restriction sites, PCR amplify the antigen-binding region of the VLR gene...
Embodiment 2
[0023] Example 2: Tumor marker CLIA detection kit
[0024] The capture reagent and detection reagent in the CLIA detection kit are replaced by the recombinant variable lymphocyte receptor (capture VLR and detection VLR) with high specificity and high affinity with tumor markers to replace the existing tumor marker CLIA detection kit The capture antibody, detection antibody, and other reagents are the same as the existing tumor marker CLIA detection kit.
[0025] The preparation method for detecting VLR is the same as that of Example 1, except that it is labeled with a chemiluminescent agent, which is different from the enzyme labeling in Example 1.
[0026] The detection method using the CLIA detection kit is the same as the prior art.
[0027] Using Example 2 of the present invention to detect CA125 samples, the intra-assay precision CV value is <4.3%, the inter-assay precision CV value is less than 7.9%; the sensitivity is less than 5 pM; the recovery rate is 103 ± 4.1%; th...
Embodiment 3
[0028] Embodiment 3: Tumor marker TRFIA detection kit
[0029] The capture reagent and detection reagent in the TRFIA detection kit are the recombinant variable lymphocyte receptors (capture VLR and detection VLR) with high specificity and high affinity with tumor markers to replace the existing tumor marker TRFIA detection kit The capture antibody, detection antibody, and other reagents are the same as the existing tumor marker TRFIA detection kit.
[0030] The preparation method for detecting VLR is the same as that of Example 1, except that it is labeled with a fluorescent agent, which is different from the enzyme labeling in Example 1.
[0031] The detection method using the TRFIA detection kit is the same as the prior art.
[0032] Using Example 2 of the present invention to detect HE4 samples, the intra-assay precision CV value is <3.7%, the inter-assay precision CV value is less than 7.1%; the sensitivity is less than 3 pM; the recovery rate is 101 ± 3.7%; the storage ...
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