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Protein eluent for dried blood spots on filter paper

A technology of eluent and protein, which is applied in the field of protein eluent, can solve the problems of reducing the sensitivity of immunoassay, taking a long time to dry blood film, and reducing the accuracy of detection, so as to improve the detection sensitivity and accuracy and shorten the detection time , low cost effect

Inactive Publication Date: 2014-06-04
BEIJING YUANDE BIO MEDICAL ENG
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] At present, the neonatal disease screening kit generally uses Tirs-HCl buffer as the protein eluent of the filter paper dried blood film. After the protein on the filter paper blood film is eluted, the immunoassay is performed. The sensitivity of immunoassay, and the presence of interfering substances, reduces the accuracy of detection
In addition, Tris-HCl takes a long time to elute the dry blood film on the filter paper. Generally, the plate needs to be washed many times, which increases the detection time.

Method used

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  • Protein eluent for dried blood spots on filter paper
  • Protein eluent for dried blood spots on filter paper
  • Protein eluent for dried blood spots on filter paper

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] Add 10‰ NP40 to 50% calf serum as the basic protein eluent, and add 0.1‰ (2-hydroxypropyl)-β-cyclodextrin and 0.1‰ (hydroxypropyl) methylcellulose As a stabilizer and an auxiliary stabilizer, respectively, and then add 1‰ of Proclin-300 as a preservative. Use a special puncher for neonatal disease screening to punch out filter paper dried blood samples with a diameter of 3 mm and TSH concentrations of 0, 3, 6, 15, 50, and 150 mIU / L, and put the punched filter paper dried blood samples into corresponding micro Add 100 μL / well of this protein eluent to the wells, and add 50 μL / well of another TSH antibody solution labeled with horseradish peroxidase (HRP) at the same time, shake the elution reaction on a 96-well plate shaker 3h, after washing the plate 5 times, add luminol luminescent solution, and measure the luminescence value with a luminometer.

[0026] Sample TSH concentration (mIU / L)

Embodiment 2

[0028] Add 50‰ NP40 to 50% calf serum as the basic protein eluent, and add 1‰ of (2-hydroxypropyl)-β-cyclodextrin and 1‰ of (hydroxypropyl) methylcellulose As a stabilizer and auxiliary stabilizer respectively, and then add 1‰ Proclin-300 as a preservative; use a special puncher for neonatal disease screening to punch out TSH concentrations of 0, 3, 6, 15, 50, and 150mIU with a diameter of 3mm / L filter paper dried blood sample, put the punched filter paper dried blood sample into the corresponding microwell, add the protein eluent 100 μL / well respectively, and also add another kind of labeled horseradish peroxidase (horseradish peroxidase ( HRP) TSH antibody solution 50 μL / well, shake and elute on a 96-well plate shaker for 3 hours, wash the plate 5 times, add luminol luminescence solution, and measure the luminescence value with a luminometer, see the table below.

[0029] Sample TSH concentration (mIU / L)

Embodiment 3

[0031] Add 65‰ NP40 to 50% calf serum as the basic protein eluent, and add 1‰ of (2-hydroxypropyl)-β-cyclodextrin and 1‰ of (hydroxypropyl) methylcellulose As a stabilizer and auxiliary stabilizer respectively, and then add 1‰ Proclin-300 as a preservative; use a special puncher for neonatal disease screening to punch out TSH concentrations of 0, 3, 6, 15, 50, and 150mIU with a diameter of 3mm / L filter paper dried blood sample, put the punched filter paper dried blood sample into the corresponding microwell, add the protein eluent 100 μL / well respectively, and also add another kind of labeled horseradish peroxidase (horseradish peroxidase ( HRP) TSH antibody solution 50 μL / well, shake and elute on a 96-well plate shaker for 3 hours, wash the plate 5 times, add luminol luminescence solution, and measure the luminescence value with a luminometer, see the table below.

[0032] Sample TSH concentration (mIU / L)

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Abstract

The invention discloses an efficient protein eluent, which comprises a surfactant, a stabilizer, an assistant stabilizer and a buffer. Preferably, the protein eluent disclosed by the invention further comprises an antiseptic. The protein eluent can effectively elute the proteins on the dried blood spots on filter paper, can remarkably reduce the interference of other substances in whole blood on immunodetection, and has the characteristics of strong specificity, low cost and good preparation easiness, therefore, the protein eluent has extensive application prospect in newborn disease screening featured by the use of protein as detection marker and of immunodetection techniques.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a protein eluent used for filter paper dried blood slices. Background technique [0002] The popularization and promotion of newborn disease screening enables early diagnosis of some congenital genetic metabolic diseases that endanger children's lives and cause children's physical and mental development disorders, and can be diagnosed before the clinical symptoms of newborns appear or when the symptoms are not obvious. Give timely treatment to avoid or reduce the irreversible damage to the various organs of the child's body. [0003] At present, many immunoassay-based detection methods, such as chemiluminescent immunoassay, time-resolved fluorescent immunoassay, enzyme-linked immunosorbent assay, and radioimmunoassay, have been applied to neonatal disease screening. However, due to the particularity of neonatal blood collection, neonatal disease screening is to detect the markers in...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/543G01N33/531
Inventor 刘宾孙旭东杨晓林张鑫李鹏翀吴晓东
Owner BEIJING YUANDE BIO MEDICAL ENG
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