HPV (human papillomavirus) immune colloidal gold diagnostic test strip, method for preparing same and detection method implemented by HPV immune colloidal gold diagnostic test strip

A technology for immunochromatographic detection and test strips, which is used in measurement devices, biological tests, material inspection products, etc., can solve the problem of not asking the market, and achieve the effects of short detection time, high specificity, and high sensitivity detection performance.

Inactive Publication Date: 2016-10-26
SUZHOU DONGNI BIOTECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0017] In terms of diagnosis of autoimmune diseases, many immunochromatographic test strips have appeared, but the immunochromatographic test strips based on HPV detection have not yet entered the market at home and abroad.

Method used

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  • HPV (human papillomavirus) immune colloidal gold diagnostic test strip, method for preparing same and detection method implemented by HPV immune colloidal gold diagnostic test strip
  • HPV (human papillomavirus) immune colloidal gold diagnostic test strip, method for preparing same and detection method implemented by HPV immune colloidal gold diagnostic test strip

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0048] Preparation of each component of the gold standard test paper of embodiment 1:

[0049] 1 Colloidal gold solution preparation: 0.01% HAuCl 4 Heat the solution to boiling, quickly add every 100mL HAuCl 4 The solution is added with an appropriate amount of reducing agent solution, the color changes from blue, then light blue, blue, then red after heating, and transparent orange-red after boiling for 7-10 minutes. Then filter with ultrafiltration or microporous membrane (0.45uM) to remove polymers and other possible impurities. The prepared colloidal gold should be pure, translucent, free of sediment and floating matter, and discard when oily matter and a large amount of black granular precipitate impurities appear on the liquid surface.

[0050] Wherein the reducing agent used can be trisodium citrate, tannic acid-trisodium citrate, white phosphorus, preferably use trisodium citrate, more preferably use 1% trisodium citrate. The glass containers used therein should be ...

Embodiment 2

[0072] Example 2 Preparation of colloidal gold-labeled anti-HPV antibody protein detection test paper 1-3

[0073] The sample pads prepared in Example 1 and dried, the binding pads, the analysis membrane, and the absorbent paper were cut into 1.7cm, 0.8cm, 2.5cm, and 1.5cm wide narrow strips with a cutting machine, according to figure 1 The way is overlapped into large slabs, and the large slabs are cut into single servings with a strip cutter. The width of each serving varies according to the jamming of the bottom plate. The preferred width of the present invention is 3mm. Assemble the test paper that has been cut for a single person into the prepared test paper card, so that the sample loading window corresponds to the sample pad of the test paper, and the result display window corresponds to the detection area and quality control area. 20~30%.

[0074] The components of gold standard test strips 1~3 are as follows:

[0075]

Embodiment 3

[0076] Embodiment 3 sample processing

[0077] Serum sample: Take 1~5mL of whole blood in a serum collection tube, let it stand for 30min~2h, centrifuge at 3000~5000g for 5~10min, and take the supernatant. According to the detection accuracy of the test strip, dilute the sample 0-100 times with the sample buffer solution, take 50-100uL dropwise into the sample hole of the test strip, and observe the result after standing for 5-20min.

[0078] Plasma sample: Take 1~5mL of whole blood, mix it in a sodium citrate or sodium heparin anticoagulant tube, centrifuge at 1000~3000g for 5~10min, and take the supernatant to get the plasma sample. According to the detection accuracy of the test strip, dilute the sample 0-100 times with the sample buffer solution, take 50-100uL dropwise into the sample hole of the test strip, and observe the result after standing for 5-20min.

[0079] Whole blood sample: Take about 50uL of fresh blood from the fingertip or earlobe, drop it into the sample ...

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Abstract

The invention discloses a quick and accurate HPV (human papillomavirus) immunochromatographic detection test strip, a method for preparing the same and a HPV detection method on the basis of immunochromatographic technologies. The immunochromatographic technologies are applied to detecting HPV antigen protein for the first time, and accordingly the HPV immunochromatographic detection test strip has high-specificity and high-sensitivity detection performance. Compared with Pasteur and HPV-DNA (deoxyribonucleic acid) detection reported in existing literature, the HPV immunochromatographic detection test strip, the method and the HPV detection method have the main advantages that the HPV immunochromatographic detection test strip and the HPV detection method are short in detection time (5-20 minutes); optional special instruments can be omitted; the HPV immunochromatographic detection test strip is easy and convenient to operate and low in detection cost, only one-step reaction is required, and operators do not need to be trained; special temperature requirements and freezing can be omitted, and accordingly the HPV immunochromatographic detection test strip is convenient to store and transport and can be stored at the room temperatures for 24 months.

Description

technical field [0001] The invention relates to the field of immunoassay detection. Specifically, the present invention relates to a preparation method and application of an immunochromatography test paper for detecting HPV, and a colloidal gold immunochromatography method using the principle of the double-antibody sandwich method. Background technique [0002] The relationship between HPV infection and cervical cancer was first proposed in the 1870s, and many epidemiological and molecular studies have confirmed the etiological link between HPV and cervical cancer beyond doubt. Bosch and Manos et al. collected cervical cancer biopsy specimens from 22 countries for PCR detection and found that human papillomavirus DNA could be detected in 99.7% of the tumors, and there was no significant difference between countries. This is the highest detection percentage among human tumor pathogenic factors reported so far, and it also shows that the association between human papillomavir...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/569G01N33/577G01N33/68
CPCG01N33/56983G01N33/577G01N33/6893G01N2333/025
Inventor 王峰王玉立包媛张建华
Owner SUZHOU DONGNI BIOTECH CO LTD
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