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Nano-selenium reagent kit for rapidly detecting HE4 and CA125

A CA125, nano-selenium technology, applied in the field of biological immunochromatography detection, can solve the problems of sensitivity, stability, difficult to control, high cost of nano-gold, complex preparation process, etc., and achieve rapid and sensitive screening, accurate detection results, and preparation. low cost effect

Active Publication Date: 2019-09-10
HENAN UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0008] 1. The cost of nano-gold is high: At present, the most commonly used marker for household lateral flow chromatography kits is nano-gold, and its preparation process and detection effect are good, but the cost of nano-gold is relatively high, and the preparation process is relatively complicated
[0009] 2. Blood sample detection has a high false positive rate and poor sensitivity: At present, lateral flow kits are relatively difficult to detect serum or blood, and are prone to false positives. Especially for the detection of double detection lines, the sensitivity and stability are more difficult to control. False positives are also higher

Method used

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  • Nano-selenium reagent kit for rapidly detecting HE4 and CA125
  • Nano-selenium reagent kit for rapidly detecting HE4 and CA125
  • Nano-selenium reagent kit for rapidly detecting HE4 and CA125

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0055] Example 1 Preparation of Nano Selenium Kit for Rapid Detection of HE4 and CA125

[0056] The Anti-HE4-Ab1, Anti-HE4-Ab2, Anti-CA125-Ab1 and Anti-CA125-Ab2 used in the kit of the present invention were purchased from MEDIX Company, and the article numbers were 4501, 4503, 4601 and 4602 in sequence.

[0057] Anti-HE4-Ab1 and Anti-HE4-Ab2 are monoclonal antibodies against different epitopes of HE4 respectively;

[0058] Anti-CA125-Ab1 and Anti-CA125-Ab2 are monoclonal antibodies against different epitopes of CA125, respectively.

[0059] (1) Preparation of nano-selenium

[0060] Take 16mL of ultrapure water and place it in a 50mL small beaker, add 1mL of 2mol / L PEG, stir at room temperature for 20min, and disperse evenly to obtain a PEG solution; dissolve 1mL of 0.1mol / L SDS in the PEG solution, and stir at room temperature for 20min , dispersed evenly, to obtain a template solution; 1mL of 0.04mol / L selenous acid was dissolved in the template solution, stirred at room t...

Embodiment 2

[0078] Example 2 Determination of the detection results of the nano-selenium kit for rapid detection of HE4 and CA125

[0079] 1. Detection method:

[0080] Add 1 drop of human whole blood or serum sample and sample treatment solution (PBS+Tween-20) to the sample pad position of the kit prepared in Example 1 of the present invention, and the sample will combine the nano-selenium on the pad with the antibody complex. Dissolved, relying on capillary action to chromatograph toward the absorbent pad, when the chromatography reaches the detection line and the quality control line, the color will be developed according to whether the sample contains the target substance or not.

[0081] 2. Result judgment:

[0082] When the quality control line C, detection line T1 and detection line T2 all develop color, the tested sample contains HE4 and sugar chain antigen CA125 above the sensitivity, such as figure 2 As shown in middle 1; when the quality control line C and the detection line...

Embodiment 3

[0083] Example Three Sensitivity Detection

[0084] In this example, the sensitivity of the kit in Example 1 is tested. Dilute the HE4 and CA125 standards with negative serum to concentrations of 100ng / mL+250U / mL, 10ng / mL+100U / mL, 2.5ng / mL+35U / mL, 0.25ng / mL+3.5U / mL Solution, the negative is the negative serum without HE4 and CA125 standards. Add 80 μL of standard solutions of different concentrations into the kit respectively. A negative result means that the T test line does not develop color, but a C quality control line develops color. A positive result means that both the T test line and the C quality control line develop color. The lowest standard solution concentration is the sensitivity of the kit, and the specific test results are as follows: image 3 shown.

[0085] image 3 Among them, 1 is a negative serum test result, which is negative, and the concentrations of 2-5 are the test results of the standard solution. It can be seen from the figure that the sensitiv...

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Abstract

The invention belongs to the field of biological immunochromatography detection methods, and specifically relates to a nano-selenium reagent kit for rapidly detecting HE4 and CA125, and a preparationmethod thereof. The reagent kit is composed of a sample pad, a bonding pad, a reaction pad, an absorbent pad and a PVC bottom plate; a detection line T1 coated with an anti-CA125 antibody A2, a detection line T2 coated with an anti-HE4 antibody B2 and a quality control line C coated with goat anti mouse IgG are arranged on the reaction pad; and rapid bedside screening and diagnosis of early ovarian cancer can be realized by observing a chromogenic reaction of a detection line. The nano-selenium immunochromatographic reagent kit prepared by the invention has the advantages of being strong in specificity, high in sensitivity, good in accuracy, simply and rapid, the required detection sample amount is small, and the reagent kit is suitable for screening of high-risk populations of ovarian cancer and home self-inspection and bedside rapid detection.

Description

technical field [0001] The invention belongs to the field of biological immunochromatography detection methods, in particular to a nano-selenium kit for rapid detection of HE4 and CA125 and a preparation method thereof. Background technique [0002] Ovarian cancer is one of the most common gynecological malignancies, with the highest mortality rate among gynecological malignancies. It also has the characteristics of high incidence, rapid disease progression, poor prognosis, and easy recurrence. The 5-year survival rate of patients with early ovarian cancer can reach 70%, but the 5-year survival rate of patients with advanced ovarian cancer is only 30%. Because the onset of ovarian cancer is hidden and the early symptoms are not obvious, clinically most patients are diagnosed at an advanced stage and miss the best time for treatment. The reasons for these situations are closely related to the lack of simple and effective early detection methods. At present, the clinical dia...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/68G01N33/574
CPCG01N33/57449G01N33/57484G01N33/689G01N2800/7028
Inventor 王志增王耀辉郭亚飞任阳光周倩文郑植张海龙马远方李淑莲
Owner HENAN UNIVERSITY
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