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Semiquantitative immunodiagnosis NGF colloidal gold test paper and preparation method thereof

A colloidal gold test paper and semi-quantitative technology, which is applied to measurement devices, instruments, scientific instruments, etc., can solve the problems of inability to provide detection results, and achieve the effects of low cost, high specificity and simple detection procedures.

Inactive Publication Date: 2009-10-14
SINOBIOWAY BIOMEDICINE +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Qualitative colloidal gold immunodiagnostic test strips cannot provide quantitative test results for the detection of the infection degree of infectious disease pathogens, endocrine disorders, whether tumor markers are higher than normal levels, and whether the intake of prohibited drugs exceeds the standard.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0023] NGF protein was used to immunize rabbits and mice to obtain anti-NGF mouse specific polyclonal antibody McAB and rabbit specific polyclonal antibody TAB respectively.

[0024] The rabbit-derived specific polyclonal antibody TAB was selected as the colloidal gold-labeled antibody, and the sodium citrate reduction method was used to prepare and determine the optimal size of the labeled nano-gold particles to be 20-50nm, and centrifuge at 9500rpm / min for 20 minutes to prepare the gold stock solution. Use PH9.0 borate buffer solution to make the labeled monoclonal antibody into a gradient solution of 5-50 μg / mL, add gold stock solution and then add 10% NaCl for stability experiment, centrifuge for 5 minutes, and measure the light absorption at 520-600nm Value (OD), when the optical density is stable, the TAB protein concentration is 10-40 μg / mL. Take TAB protein solution with a concentration of 10-40 μg / mL, add gold stock solution, and then add PEG to make colloidal gold st...

Embodiment 2

[0031]The difference from Example 1 is that three NGF strips of 10 μg / mL, 100 μg / mL, and 50 μg / mL were sprayed laterally on the semi-quantitative area from bottom to top; when used, if only one band is displayed in the semi-quantitative area, it indicates that the mouse submandibular gland The NGF protein present in the homogenate supernatant sample is less than 100 μg / mL level; two bands show that the NGF protein present in the mouse submandibular gland homogenate supernatant sample is greater than or equal to 100 μg / mL; if the 100 μg / mL band and If there is little difference in the horizontal color on the left side, the NGF protein in the homogenate supernatant of the mouse submandibular gland is close to 100 μg / mL; 100-150 μg / mL.

Embodiment 3

[0033] The difference from Example 1 is that the semi-quantitative area is sprayed horizontally from bottom to top in eight strips of 10 μg / mL, 150 μg / mL, 120 μg / mL, 100 μg / mL, 80 μg / mL, 60 μg / mL, 40 μg / mL and 20 μg / mL NGF band; when used, if only one band is displayed in the semi-quantitative area, it indicates that the NGF protein present in the homogenate supernatant sample of the mouse submandibular gland is less than 100 μg / mL; if there are multiple bands in the semi-quantitative area, there are more bands The higher the NGF protein content in the mouse submandibular gland homogenate supernatant: two bands show that the NGF protein present in the mouse submandibular gland homogenate supernatant sample is greater than or equal to 100 μg / mL; if the 100 μg / mL band and the left If there is little difference in color at the side level, the NGF protein in the homogenate supernatant sample of the mouse submandibular gland is close to 100 μg / mL; if three bars show, the concentrati...

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Abstract

The invention discloses a semiquantitative immunodiagnosis NGF colloidal gold test paper and a preparation method thereof. The semiquantitative immunodiagnosis NGF colloidal gold test paper is separated into a first sample absorption area, a second immobilization colloidal gold mark NGF polyclonal antibody area, an third immobilization another NGF polyclonal antibody area, a fourth immobilization NGF area and a fifth water absorption area from bottom to top, wherein spaced narrow slits are symmetrically distributed on both sides of the third immobilization another NGF polyclonal antibody area and the fourth immobilization NGF area. The semiquantitative immunodiagnosis NGF colloidal gold test paper is characterized in that the fourth immobilization NGF area comprises a clutter signal removing stripe and 2 to 7 NGF stripes with degressive concentration. Through the semiquantitative immunodiagnosis NGF colloidal gold test paper, the color of nanometer colloidal gold markers can be rapidly visually inspected after the nanometer colloidal gold markers are combined with samples and a detected semiquantitative result can be judged according to the comparison of color at the left sides and the right sides of testing areas and the number of the strips of a right side area by adopting an immunochromatographic principle. The semiquantitative immunodiagnosis NGF colloidal gold test paper has the characteristics of reacting rapidly and rapidly judging if the samples have objects to be tested and the amount of the content of the objects to be tested.

Description

technical field [0001] The invention relates to a colloidal gold test paper and a preparation method thereof, in particular to a colloidal gold test paper for semi-quantitative detection of NGF content and a preparation method thereof. Background technique [0002] The detection of nerve growth factor (NGF) mainly uses high-performance liquid chromatography (HPLC) and enzyme-linked immunosorbent assay (ELISA). The HPLC method is sensitive and accurate, and the lower limit of detection can reach 0.25ng / mL. However, because this method requires expensive instruments such as HPLC, sample pretreatment is cumbersome, and the analysis speed is slow. Specially trained personnel can operate, the detection cost is high, the detection of batch samples is difficult, and the application range is narrow. ELISA method is currently the most researched NGF detection method. It has high sensitivity, large detection capacity, simple sample pretreatment, shorter analysis time and lower cost t...

Claims

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Application Information

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IPC IPC(8): G01N33/558G01N33/543
Inventor 邹宇飞熊玲媛马凌燕付永超张键荣付晓妍
Owner SINOBIOWAY BIOMEDICINE
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