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Method for ovarian cancer screening based on ovarian cancer marker and logic gate operation

A technology for ovarian cancer and tumor markers, applied in the field of biomedicine, can solve the problems of large antibody molecules in detection methods, and achieve the effects of improving detection sensitivity, high sensitivity, and strong specificity

Active Publication Date: 2019-11-08
JIANGNAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] At present, technical means such as radioimmunoassay, immunoradiometric assay, enzyme-labeled immunoassay, chemiluminescence immunoassay, and time-resolved fluorescence immunoassay are usually used to detect the tumor marker content in serum of patients. The principle is based on the combination of antigen and antibody, and the detection method is relatively limited mainly because the antibody molecule is large and immunogenic

Method used

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  • Method for ovarian cancer screening based on ovarian cancer marker and logic gate operation
  • Method for ovarian cancer screening based on ovarian cancer marker and logic gate operation
  • Method for ovarian cancer screening based on ovarian cancer marker and logic gate operation

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0055] Example 1: "NOR" logic gate

[0056] (1) Formation of hybrid strands: use PBS buffer (2mmol·L -1 K H 2 PO 4 ,10mmol·L -1 Na 2 HPO 4 ,2.7mmol·L -1 KCl, 137mmol·L -1 NaCl, pH7.4) to dilute TemDNA, CEA-apt, CA125-apt to 1 μmol L -1 , after dilution, take 15 μL each and mix evenly, react at 95°C for 5 minutes, and react at 25°C for 2 hours to form a stable hybrid chain structure;

[0057] Wherein the CEA-apt sequence is shown in SEQ ID NO.1 (5'-GGGTTGGGTTTATACCAGCTTTATTCAATT-3'); the CA125-apt sequence is shown in SEQ ID NO.2 (5'-TAATACGACTCACTATAGGGAGACAAGAATAAACGCTCAATTTGGGTAGGG-3'); the TemDNA sequence is shown in SEQ ID NO.3 (5'-TTGAATAAGCTGGTATTTTGAGCGTTTATTCTTGTCTC CCTATAGTGAGTCGTA-3');

[0058] (2) DNA-marker protein incubation: add 2 μL of CEA and CA125 samples of different concentrations to the reaction system in step (1), and incubate at 37°C for 1 hour;

[0059] (3) Formation of G-quadruplex-heme complex: add 423.5μL workingbuffer (150mmol·L -1 NH 4 C...

Embodiment 2

[0062] Example 2: "AND" logic gate

[0063] (1) Formation of complementary double strands of apt-cDNA: dilute CEA-cDNA, CA125-cDNA, CEA-apt, CA125-apt to 2 μmol L with PBS buffer -1 After dilution, take 75 μL each and mix evenly, heat at 95°C for 5 minutes, and hybridize at room temperature for 2 hours to form a stable complementary double-stranded structure;

[0064] Wherein the CEA-apt sequence is shown in SEQ ID NO.1 (5'-GGGTTGGGTTTATACCAGCTTTATTCAATT-3'); the CA125-apt sequence is shown in SEQ ID NO.2 (5'-TAATACGACTCACTATAGGGAGACAAGAATAAACGCTCAATTTGGGTAGGG-3'); the CEA-cDNA sequence is shown in SEQ ID NO.4 (5'-Biotin-AAAAAAAAAAAAAAAATTAATTGAATAAGCT-3'); CA125-cDNA sequence is shown as SEQ ID NO.5 (5'-TGTCTCCCTATAGTGAGTCGTATTATTAAAAAAAAAAAAAAA-Biotin-3');

[0065](2) Construction of the magnetic bead-apt-cDNA complex: place the magnetic bead bottle on a vortex shaker for 20 seconds, and resuspend the magnetic beads modified with streptavidin. Take 100 μL of the magnetic b...

Embodiment 3

[0071] Example 3: Detection results of ovarian cancer screening by "NOR" logic gate operation

[0072] The detection of tumor markers is divided into 5 groups, A(0,0), P(0,0), (0,1), (1,0), (1,1). Five groups contained different concentrations of CEA and CA125. Among them, A(0,0) group has no CEA and CA125; P(0,0) group contains 5ng·mL -1 CEA and 35U·mL -1 CA125, that is, the content of CEA and CA125 under normal physiological indicators; (1,0) group contains 40ng·mL -1 CEA and 35U·mL - 1 CA125, that is, pathological CEA content and normal physiological CA125 content; (0,1) group contains 5ng·mL -1 CEA and 100U·mL - 1 CA125, that is, normal physiological CEA content and pathological CA125 content; (1,1) group contains 40ng·mL -1 CEA and 100U·mL - 1 CA125, that is, the content of CEA and CA125 are pathological content. After forming a stable hybrid chain structure, add the concentration of CEA and CA125 to the detection solution respectively according to the concentr...

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Abstract

The invention discloses a method for ovarian cancer screening based on an ovarian cancer marker and a logic gate operation, and belongs to the technical field of biomedicine. The protein-nucleic acidrecognition technology, the signal amplification technology and a logic gate are combined, aptamers of CEA and CA125 are used for target recognition, signal transformation and amplification are performed through a G-quadruplex-heme complex, and a method for simultaneous detection of tumor markers is established. According to the method, an optical biosensor with high sensitivity and strong specificity is constructed for achieving the joint detection of the CEA and the CA125, and an ovarian cancer screening result is provided based on the logic gate; through the combination of optical detectionand the signal amplification technology, the detection sensitivity is improved; the universality of the detection principle makes the method also suitable for detection of different objects based ondifferent protein recognition elements.

Description

technical field [0001] The invention relates to a method for screening ovarian cancer based on ovarian cancer markers and logic gate operations, belonging to the technical field of biomedicine. Background technique [0002] Ovarian cancer is a malignant tumor that occurs in the ovarian tissue. It ranks third among female reproductive system malignant tumors and is the tumor with the highest gynecological mortality rate. Therefore, early detection, early diagnosis and early treatment are extremely important for the rehabilitation of ovarian cancer patients. . When the early tumor is limited to the ovary, ultrasonography is difficult to diagnose, and the increase of serum tumor markers can precede the imaging changes and clinical symptoms for several months, which has certain clinical significance for early diagnosis. Ovarian cancer has a variety of tumor markers, of which only carbohydrate antigen 125 (CA125) is clinically recognized and widely used in ovarian tumors, and th...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/574G01N33/543G01N21/31
CPCG01N21/31G01N33/54326G01N33/57449
Inventor 周楠迪姚斌斌荆丞李茂林田亚平
Owner JIANGNAN UNIV
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