A lupus anticoagulant detection reagent

A lupus anticoagulant and detection reagent technology, which is applied in the field of clinical detection, can solve problems such as difficult matching, increased false positive rate or false negative rate of test results, etc., and achieves the effects of easy production, improved accuracy, and improved sensitivity

Active Publication Date: 2019-03-05
NINGBO ACCUTECH BIOSCI LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Therefore, it is difficult to match the components of the screening reagent in the form of lyophilized powder and the confirm

Method used

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  • A lupus anticoagulant detection reagent
  • A lupus anticoagulant detection reagent
  • A lupus anticoagulant detection reagent

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0024] The screening of embodiment 1 synthetic phospholipids

[0025] Weigh an appropriate amount of synthetic phosphatidylserine, phosphatidylcholine, phosphatidylglycerol, phosphatidylethanolamine, and cholesterol, and dissolve them in chloroform in different proportions (accounting for the mass percentage of synthetic phospholipids) (see Table 1 for details), mix well, and use Blow dry with nitrogen, then vacuum to dry residual chloroform. Add a certain amount of distilled water and stir at room temperature for 2 hours to prepare a phospholipid liposome solution. Store at -20°C for later use.

[0026] Table 1

[0027] Phospholipid component

Phosphatidylserine

Phosphatidylcholine

Phosphatidylethanolamine

Phosphatidylglycerol

cholesterol

Phospholipid 1

10％

30％

40％

15％

5％

Phospholipid 2

10％

35％

35％

15％

5％

Phospholipid 3

10％

45

35％

10％

5％

Phospholipid 4

5％

30％

40％

...

Embodiment 2

[0035] Embodiment 2 The preparation of the reagent of lupus anticoagulant of the present invention

[0036] (1) Preparation of screening reagents

[0037] Prepare 50mM Tris buffer, add 100mM sodium chloride, add appropriate amount of silica (final concentration is 0.28%), add stabilizers and preservatives, so that the solution contains 1% BSA by mass volume, 1% PEG6000, 0.02% BHT , 0.05% sodium azide, adjust the pH to 7.35, add an appropriate amount of the prepared phospholipid 4 solution in Example 1, so that the final concentrations are 10 μM, 40 μM, 160 μM and mix well. Set up the program according to the instructions of the instrument, and use the prepared screening reagent to test the normal plasma and lupus anticoagulant positive plasma on the Sysmex CA1500 hemagglutination analyzer. The test parameters are: take 50 μl of the screening reagent and add it to the reaction tube. , add 50 μl of blood sample, incubate at 37°C for 3 minutes, add 50 μl of calcium chloride solu...

Embodiment 3

[0046] Embodiment 3 Stability

[0047] Prepare screening reagents and diagnostic reagents according to Example 2, place the reagents at 4°C, incubate at 37°C, and take samples for detection every 7 days. The results are shown in Table 5.

[0048] table 5

[0049]

[0050] The results showed that the detection results of the lupus inhibitor detection reagent did not change much after being stored at 37°C for 28 days, and the stability was very good.

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Abstract

The invention discloses a lupus anticoagulant detection reagent which includes a screening reagent and a diagnosis reagent. Both of the two reagents include a buffer solution, an activating agent silica, and synthetic phospholipid, wherein the content of the synthetic phospholipid is low in the screening reagent but is high in the diagnosis reagent. The synthetic phospholipid is composed of phosphatidyl serine, phosphatidyl choline, phosphatidyl ethanolamine, phosphatidyl glycerol and cholesterol according to the mass ratio of 5:30:40:15:10. The lupus anticoagulant detection reagent is high in sensitivity and is a liquid reagent; compared with a DRVVT method, the lupus anticoagulant detection reagent is very small in inter-batch difference and is more easy to produce.

Description

technical field [0001] The invention relates to the technical field of clinical detection, in particular to a reagent for detecting lupus anticoagulant. Background technique [0002] Antiphospholipid antibodies are a collective term for a class of autoantibodies that bind to phospholipids or proteins that form complexes with phospholipids. Antiphospholipid antibodies have different names depending on which phospholipid or phospholipid-binding protein the antibody recognizes. For example, anti-cardiolipin antibodies, anti-β2glycoprotein Iantibodies, phosphatidylserine-dependent antiprothrombin antibodies, lupus anticoagulant (lupus anticoagulant). Lupus anticoagulants were first discovered in lupus patients and later found to be present in large quantities in many non-lupus patients. The mechanism of lupus anticoagulant formation is unclear, and it may be related to medication, such as chlorpromazine, procainamide, azathioprine, etc. Lupus anticoagulant can cause antiphos...

Claims

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Application Information

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IPC IPC(8): G01N33/86
CPCG01N33/86
Inventor 赖增祖叶欣杨奎东徐健邬荣杰
Owner NINGBO ACCUTECH BIOSCI LTD
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