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Reagent for specific detection of fungi in serum and preparation method of reagent

A kind of specific and reagent technology, which is applied in the field of reagents and its preparation for specific detection of fungi in serum, can solve the problems of complex process, loss and low yield, and achieve the effect of avoiding false positive results

Inactive Publication Date: 2018-08-10
福州启新生物技术有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Therefore, in the current technology, when using LAL to detect serum fungi, the LAL must be chromatographically purified to remove factor C or / and B factor, but the process is more complicated and the yield is low, factor G and other components Points will also cause a certain loss

Method used

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  • Reagent for specific detection of fungi in serum and preparation method of reagent
  • Reagent for specific detection of fungi in serum and preparation method of reagent
  • Reagent for specific detection of fungi in serum and preparation method of reagent

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0067] 1. Limulus reagent preparation:

[0068] 1.1 Cell shearing:

[0069] Preparations: Prepare the tools required for cutting the cells after removal of bacterial endotoxin on the 100-level operating table. Pre-freeze the glass vials to be loaded with cells in a freezer below 4°C. Pre-freeze the disintegration solution to be used in a refrigerator at 0-4°C for 30 minutes. Take about 40-50g of qualified cell raw material each. Pick 2 beakers and cover the sides with ice.

[0070] Cutting cells: Take a few branches of cells, carefully dig out the cells in the container with a medicine spoon, pick up the cells with tweezers, and cut the cells into a beaker with scissors. It is required to cut 25g cells into more than 10 small pieces of cell fragments. Clamp the cut cells in the beaker into the pre-frozen glass bottles, add the pre-frozen disintegration solution into the glass bottles containing the cells at a ratio of 1:8, and shake each glass bottle vigorously , to dispe...

Embodiment 2-5

[0081] The difference between Examples 2-5 and Example 1 is that in the preparation step of the detection reagent, the polymyxins added are respectively polymyxins A, B, C, and D.

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Abstract

The invention provides a reagent for specific detection of fungi in serum. The reagent comprises polymyxin and tachypleus amebocyte lysate. According to the technical scheme, polymyxin is added in thetachypleus amebocyte lysate. The polymyxin contains positive-charged free amino groups, which can combine with negative-charged phosphate groups in phosphatide of Gram-negative bacteria lipopolysaccharides (LPS) which is endotoxin to make endotoxin inactive. The inactive lipopolysaccharides cannot react with a C factor in tachypleus amebocyte lysate, so that a C reaction is inhibited, and a falsepositive result in fungi detection is avoided. Cross reaction doesn't happen between polymyxin and other components in tachypleus amebocyte lysate or detected serum.

Description

technical field [0001] The invention relates to the field of biochemistry, in particular to a reagent for specifically detecting fungi in serum and a preparation method thereof. Background technique [0002] The main components of Limulus reagent include factor C (Factor C, FC), factor B (Factor B, FB), factor G, proclotting enzyme (Proclotting enzyme), proclotting protein (or chromogenic substrate), divalent cations, and buffer salts. Wait. The basic principle of its enzyme cascade reaction has been widely studied, including two agglutination reaction pathways: one is the factor G (FG) pathway mediated by 1,3-β-D-glucan, and the activated factor G will coagulate The zymogen (Proclotting enzyme) is converted into clotting enzyme (clotting enzyme), and the coagulation enzyme hydrolyzes the chromogenic substrate (Boc-Leu-Gly-Arg-pNA) to release p-nitroaniline (pNA). The nitriding reaction produces a rose-red azo substance with an absorption peak at 545nm. The other is the e...

Claims

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Application Information

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IPC IPC(8): G01N33/68G01N33/579G01N33/569
CPCG01N33/56961G01N33/579G01N33/68G01N2333/37
Inventor 丁友玲陈晓佳李红玲
Owner 福州启新生物技术有限公司