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Lipoarabinomannan detection sensitizer and its composition and kit

A mannan and composition technology, applied in the field of detection compositions and detection kits, can solve problems such as difficulty in achieving sensitivity, and achieve the effect of improving high sensitivity, sensitivity and/or specificity

Active Publication Date: 2022-07-01
SHANGHAI RONGSHENG BIOLOGICAL PHARM CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, in some cases, it is difficult to achieve the goal of improving sensitivity through these two technical solutions

Method used

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  • Lipoarabinomannan detection sensitizer and its composition and kit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] Example 1 Preprocessing samples

[0035] Take 1ml of the positive sample and distribute it into 10 centrifuge tubes, each with 100ul, and add an appropriate amount of the above-mentioned sample treatment solution to each centrifuge tube to make the pH 1-2. The resulting mixture is milky white due to denaturation of the protein. The denatured protein was centrifuged at 13,000 rpm for 5 minutes in a high-speed centrifuge, the upper 75 μl clear liquid was discarded, and the pH was adjusted to 6.0-8.0 with 2M potassium carbonate solution. The sample was cooled to 4°C for 30 minutes to increase the precipitation rate, and the solution was returned to room temperature (25°C) before analysis for use.

Embodiment 2

[0036] Example 2 Preparation of lipoarabinomannan

[0037] Lipoarabinomannan can be obtained commercially (eg, Hangzhou Huakui Jinpei Biotechnology Co., Ltd.), or prepared by itself (see: CN2010105436856).

Embodiment 3

[0038] Example 3 Preparation of coated reaction plate

[0039] Preparation of coating solution: take a 200mL reagent bottle, add 0.5mL of sodium carboxymethylcellulose mother solution, add 10mL of Tris-HCL mother solution, add 10mL of trehalose mother solution, and add pure water to 100g.

[0040] Take a 15mL centrifuge tube, and use the above-mentioned coating dilution solution according to the appropriate concentration to lipoarabinomannan antigen and polypeptide (the polypeptide shown in SEQ ID No 1, or the polypeptide shown in SEQ ID No 1 and SEQ ID No 2. The peptide or the peptide shown in SEQID No 3 with a molar ratio of 2 to 5:1) was diluted to prepare a coating mixture of a suitable concentration, and an enzyme-linked immunoluminescence plate was taken, 125 μl per well, 2 ℃ ~ 8 ℃ overnight; wash twice with washing solution; block the ELISA plate with 5% BSA blocking solution, 220 μl per well, overnight at 2 ℃ ~ 8 ℃; blot dry the blocking solution in the reaction well, ...

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Abstract

A sensitizer for the detection of Mycobacterium tuberculosis lipoarabinomannan antibody, which is the polypeptide shown in SEQ ID No 1, or the polypeptide shown in SEQ ID No 2 and the polypeptide shown in SEQ ID No 3 and SEQ ID No. 3 respectively. The polypeptide shown in No. 1 forms a composition, which is then co-coated with Mycobacterium tuberculosis lipoarabinomannan antigen to realize the detection of Mycobacterium tuberculosis. The present invention adopts the way of co-coating lipoarabinomannan antigen and polypeptide (or composition), realizes the specific detection of Mycobacterium tuberculosis antibody, has high sensitivity, and is suitable for in vitro detection.

Description

technical field [0001] The present invention relates to a polypeptide auxiliary, in particular to a sensitizer for improving detection sensitivity and specificity, which is used for the detection of lipoarabinomannan, as well as a detection composition and a detection kit. Background technique [0002] Tuberculosis is an infectious disease that has plagued humans for thousands of years, and an estimated one-quarter of the adult population in nineteenth-century Europe died from tuberculosis. Thanks to improvements in living standards in the twentieth century and the discovery of antibiotics, by the 1950s, tuberculosis had been nearly eradicated in industrialized countries. However, tuberculosis remains rampant as a recurring infectious disease. At present, it is estimated that one in three people in the world's population is infected with tubercle bacillus (latent tuberculosis). The total number of new-onset tuberculosis patients in the world is said to be 9.4 million, and ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/535G01N33/545G01N33/569
CPCG01N33/535G01N33/545G01N33/569
Inventor 盛盼龙朱绍荣
Owner SHANGHAI RONGSHENG BIOLOGICAL PHARM CO LTD