Test strip and detection method for detecting phospholipase A2 receptor (PLA2R) antibody

A technology for detecting test strips and antibodies, applied in biological testing, measuring devices, material inspection products, etc., can solve the problems of cumbersome operation process, impossible quantitative detection, weak color signal, etc., and achieve strong color signal and fast color development speed , good effect of specificity

Active Publication Date: 2018-07-13
SHENZHEN BLOT BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0005] In view of this, the object of the present invention is to provide a quantitative, accurate and convenie

Method used

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  • Test strip and detection method for detecting phospholipase A2 receptor (PLA2R) antibody
  • Test strip and detection method for detecting phospholipase A2 receptor (PLA2R) antibody
  • Test strip and detection method for detecting phospholipase A2 receptor (PLA2R) antibody

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0041] Embodiment 1, the making of the fluorescent test strip of the detection PLA2R antibody of the present invention

[0042] 1. Preparation of avidin (SA) fluorescent microsphere conjugates and DNP-BSA fluorescent microsphere conjugates

[0043] 1.1 Preparation of 50mM pH7.0 MES solution

[0044] 1.2 Preparation of 50mM pH6.5 MES solution

[0045] 1.3 Dialyze the avidin and DNP-BSA to be coupled with 50mM pH7.0 MES buffer three times respectively, and temporarily store them at 4°C for later use.

[0046] 1.4 Activation of microspheres: 20 μl of 200 nm microspheres, add 50 mM pH6.5 MES solution, centrifuge and wash twice at 12000 r / min, add 0.25 μg NHS (N-hydroxysuccinimide) and 0.25 μg EDC (1 -(3-Dimethylaminopropyl)-3-ethylcarbodiimide), mixed and reacted at room temperature for 40min.

[0047] 1.5 Coupling of microspheres: After the above-mentioned activated microspheres were centrifuged at 12000r / min to remove the supernatant, they were centrifuged and washed twice wi...

Embodiment 2

[0059] Embodiment 2, the performance test of the fluorescent test strip of the PLA2R antibody of the present invention

[0060] Add 60 μl of serum to each sample in the following tests, and the instrument reads the test results in 7 minutes.

[0061] 1. Determination of the test strip critical value of the present invention: by detecting 220 normal human sera, the average value of the anti-PLA2R antibody concentration is 12.8RU / ml and the standard deviation is 1.17RU / ml, with the average value plus 3 times the standard The sum of the differences is used as the critical value, and the critical value is 16.31RU / ml.

[0062] 2. Determination of the precision of the test strip of the present invention: select 3 parts of quality control serum with high, medium and low values ​​(high value control target value: 300RU / mL, medium quality control target value: 70RU / mL, low value control target value Value: 30RU / ml), in the same test, each repeated measurement 10 times, calculate the a...

Embodiment 3

[0077] Embodiment 3, compared with the Chinese patent whose application number is 201510245291.5

[0078] 1. Select 10 weakly positive quality control sera to test, respectively adopt the test strip of the present invention and the test strip of the Chinese patent application number 201510245291.5 to detect the PLA2R antibody concentration in the serum, and read the results at 7 minutes and 15 minutes respectively. See Table 4.

[0079] Table 4 Comparison of detection results of weak positive quality control serum at different reading times

[0080]

[0081] The results show that the 7-minute interpretation result of the test strip with the application number of the Chinese patent 201510245291.5 is quite different from the theoretical concentration. The 7-min and 15-min judgment results of the test strip according to the present invention are consistent with the theoretical concentration, and the test result is more accurate.

[0082] 2. Select 15 negative clinical samples...

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Abstract

The invention belongs to the technical field of biological detection, and discloses a test strip and a test method for detecting a phospholipase A2 receptor (PLA2R) antibody. The test strip for detecting the PLA2R antibody comprises a sample pad, a conjugate pad 1, a conjugate pad 2, an NC film and absorbent paper which are sequentially in lap joint with a viscous bottom plate, wherein the conjugate pad 1 is sprayed with a PLA2R biotinylated conjugate; the conjugate pad 2 is sprayed with an SA fluorescent microsphere conjugate and a DNP-BSA fluorescent microsphere conjugate; the NC film is enveloped with a capture body as a detection line and is enveloped with an anti-DNP-BSA antibody as a quality control line. The test strip for detecting the PLA2R antibody by using a capture method has afaster color development speed, and only 7 minutes is needed for sample feeding to result reading, so that the detection time is shorter; the test strip is stronger in coloring signals, the missing detection of weak positive does not easily occur, and the specificity is better; the test strip and the test method can be used for quantitatively, accurately and conveniently testing the content of the PLA2R antibody in human serum, plasma and whole blood.

Description

technical field [0001] The invention belongs to the technical field of biological detection, and in particular relates to a PLA2R antibody detection test strip and a detection method. Background technique [0002] Membranous nephropathy (MN) is one of the most common pathological types of nephrotic syndrome in adults, and its characteristic pathological change is the deposition of a large number of immune complexes on the epithelial side of glomerular capillary loops. According to the etiology, MN can be divided into two categories: idiopathic membranous nephropathy (IMN) and secondary membranous nephropathy (SMN). IMN is a chronic glomerular inflammatory disease, most of which are associated with anti-phospholipase A2 receptor antibodies, which bind to the corresponding antigens on podocytes to form immune complexes in situ, and then through bypass The pathway activates complement, forms C5b-9 membrane attack complex, damages podocytes, and destroys the glomerular filtrati...

Claims

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Application Information

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IPC IPC(8): G01N33/68G01N33/558
CPCG01N33/558G01N2800/347G01N33/6893G01N33/54313G01N33/533G01N33/52G01N33/54386
Inventor 熊祖应张永顶马伟民张大准王洪涛
Owner SHENZHEN BLOT BIOTECH
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