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Lysophosphatidylcholine detection kit and lysophosphatidylcholine detection method

A detection kit, phosphatidylcholine technology, applied in the field of medical detection, to achieve the effect of reducing impurity interference, accurate qualitative and simple operation

Pending Publication Date: 2020-07-21
WUHAN ADICON CLINICAL LAB
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

A defect in the ABCD1 gene is responsible for the disease

Method used

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  • Lysophosphatidylcholine detection kit and lysophosphatidylcholine detection method

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Experimental program
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Effect test

Embodiment 1

[0015] In this embodiment, the lysophosphatidylcholine (LPC) at C20, C22, C24 and C26 in the dried blood spot of the sample is specifically measured by liquid chromatography-tandem mass spectrometry. The specific detection steps are as follows:

[0016] 1) Standard product preparation: Dilute the following standard products with water / methanol / acetonitrile mixture: C18:0, C20:0, C22:0, C24:0. (C18:0 represents stearic acid with 18 carbons and 0 unsaturation) Specifically, water 10%; methanol 60%; acetonitrile 30%.

[0017] 2) Standard blood sheet preparation: take normal goat or bovine whole blood cells, centrifuge at 5000rpm for 10 minutes, remove plasma, add 2 times the volume of PBS, mix with a mixer and let stand for 5 minutes, repeat this step twice to wash the blood cells. Add the blood cells that will be washed in the end, use the same volume of PBS, add the appropriate standard mixture and BHT reagent, and mix well. Take 50ul drops into the filter paper as a standard...

Embodiment 2

[0023] Embodiment 2 contrast experiment to detection background signal

[0024] According to the method of Example 1, common treatment liquid extraction (methanol) (control 1) and mixed treatment (methanol / isopropanol / acetonitrile / internal standard mixed solution 3v:3v:3v:1v) were used for treatment (the present invention). Samples 1 to 5 were extracted and determined, and the test results are shown in Table 1. The extract reagent of the invention can protect and quickly extract the required substances and reduce the interference of impurities.

[0025] Table 1

[0026] Methanol treatment background signal Background signal from mixed reagents sample 1 3120 257 sample 2 5010 301 sample 3 6190 660 sample 4 980 310 Sample 5 1770 270

Embodiment 3

[0027] Quantitative determination of embodiment 3 clinical samples

[0028] According to the method of Example 1, LC-MSMS quantitative determination was performed on the standard substance and samples 1 to 6. The measurement results are shown in Table 2. It can be proved from the table that the method of the present invention can carry out quantitative determination on the samples. The internal standard reference can be qualitative.

[0029] Table 2

[0030]

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Abstract

The invention provides a lysophosphatidylcholine detection kit and a lysophosphatidylcholine detection method. The lysophosphatidylcholine of C20, C22, C24 and C26 in dry blood spots is specifically measured through liquid chromatography-tandem mass spectrometry. The detection kit can protect and rapidly extract required substances and reduce impurity interference. The detection method is simple and rapid to operate and high in throughput, and a quantitative and qualitative method with high specificity, accuracy and sensitivity is established.

Description

technical field [0001] The invention belongs to the field of medical detection, in particular to a rapid detection kit and detection method for lysophosphatidylcholine. [0002] technical background [0003] Peroxisomes are organelles present in all human cells except mature red blood cells. They perform essential metabolic functions, including β-oxidation of very long-chain fatty acids (VLCFAs), α-oxidation of phytanic acid, and biosynthesis of plasmalogens and bile acids. Peroxisome disorders include 2 major subgroups: disorders of peroxisome biogenesis and defects in monoperoxisomal enzyme transporters. Peroxisome biogenesis defects such as Zellweger spectrum syndrome are characterized by defective assembly of entire organelles, whereas in single enzyme transporter defects such as X-linked adrenoleukodystrophy (X-ALD), organelles are intact but Certain functions are broken. These disorders are clinically diverse and vary in severity, ranging from neonatal lethal to mild...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N30/88G01N30/06
CPCG01N30/88G01N30/06
Inventor 陈坚
Owner WUHAN ADICON CLINICAL LAB