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

Active Publication Date: 2014-12-03
LIAONING NORMAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, so far there is no report on the application of variable lymphocyte receptor (VLR) in jawless vertebrates instead of capture antibodies and detection antibodies in tumor marker detection kits

Method used

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Examples

Experimental program
Comparison scheme
Effect test

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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Abstract

The invention discloses a tumor marker detection kit based on variable lymphocyte receptors (VLR) and a method for preparing the same, wherein the tumor marker detection kit is high in detection sensitivity, convenient to store and transport, long in period of validity and low in production cost. Just like the existing tumor marker detection kit, the tumor marker detection kit provided by the invention also contains a trapping reagent and a detection reagent; however, the difference is that the trapping reagent and the detection reagent are recombinant variable lymphocyte receptors having high specificity and high affinity to the tumor marker, respectively; and therefore, the clinical detection of the tumor marker is more simple and convenient and early diagnosis of cancer is more accurate and effective.

Description

technical field [0001] The invention relates to a tumor marker detection kit and a preparation method thereof, in particular to a variable lymphocyte receptor (VLR)-based tumor marker with high detection sensitivity, convenient storage and transportation, long validity period and low production cost Detection kit and preparation method thereof. Background technique [0002] At present, the detection of tumor markers in serum samples of patients has been widely used in the early diagnosis of cancer, and at the same time plays an important auxiliary role in the monitoring and prognosis analysis of cancer during treatment. There are many kinds of tumor markers. The common tumor markers are divided into embryonic antigens, carbohydrate antigens, natural autoantigens, cytokeratin, tumor-related enzymes and hormones, etc. Some tumor markers have specific effects on the detection of different types of cancer. Broad-spectrum, and some have high sensitivity and specificity for speci...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/574C12N15/12C12N15/63C12N1/19C07K14/705
Inventor 李庆伟韩英伦于涛
Owner LIAONING NORMAL UNIVERSITY
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