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A chlamydiae pneumoniae IgM chemiluminescent immunoassay kit and a preparing method thereof

A chemiluminescence immunity, Chlamydia pneumoniae technology, applied in chemiluminescence/bioluminescence, analysis by chemical reaction of materials, measurement devices, etc., can solve the problems of narrow detection linear range, low detection sensitivity, and high detection cost, reaching The effect of high accuracy, high detection sensitivity and easy operation

Inactive Publication Date: 2017-02-15
SHENZHEN YHLO BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0012] The current Chlamydia pneumoniae IgM detection technology has the following disadvantages: high detection cost, low detection sensitivity, narrow detection linear range, low reproducibility, inability to quantify, complicated operation, etc.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0021] Example 1: Preparation method of Chlamydia pneumoniae IgM chemiluminescent immunoassay kit

[0022] (1) Preparation of nano magnetic beads coated with IgM monoclonal antibody of Chlamydia pneumoniae:

[0023] Take 50mg of carboxylated magnetic particle (particle size: 0.05-1um) suspension, magnetically separate to remove the supernatant, resuspend with 0.02 M, pH 5.5 MES buffer, add 0.5-2mL of newly prepared 10 mg / mL EDC Aqueous solution, activate carboxyl groups on the surface of magnetic beads, add 3-5 mg Chlamydia pneumoniae IgM monoclonal antibody, suspend at room temperature for 2-10 h, magnetically separate, remove supernatant, buffer with 0.1 M Tris pH 8.0 containing 2% BSA The solution was resuspended to 1 mg / mL to obtain Chlamydia pneumoniae IgM monoclonal antibody-coated magnetic particles, which were divided into 5 mL bottles and stored at 4°C for later use.

[0024] (2) Preparation of acridinium esters labeled with IgM derivatives of Chlamydia pneumoniae: ...

Embodiment 2

[0032] Embodiment 2: Chlamydia pneumoniae IgM chemiluminescent immunoassay method:

[0033] The present invention uses a fully automatic chemiluminescence immunoassay analyzer as a detection tool, and the methodological mode of the present invention is a competitive method, that is, the instrument sequentially adds 20 uL of samples, 50 uL of magnetic particles coated with IgM monoclonal antibody of Chlamydia pneumoniae, and 50 uL of The acridinium ester coated with Chlamydia pneumoniae IgM was reacted for 10 minutes, then magnetically separated, and the instrument sent the reaction mixture into the dark room, followed by adding 50uL chemiluminescence pre-excitation solution and 50uL chemiluminescence excitation solution for luminescence reaction, and finally recorded the luminescence intensity , Calculate the Chlamydia pneumoniae IgM content of the tested sample from the standard curve.

Embodiment 3

[0034] Example 3: Performance evaluation of Chlamydia pneumoniae IgM chemiluminescence immunoassay kit

[0035] Sensitivity detection:

[0036] The sensitivity reference product in the serum plate was tested, which met the technical requirements of the kit.

[0037] Accuracy test:

[0038] The positive reference and negative reference in the serum plate were tested, and the compliance was 100%.

[0039] Precision determination:

[0040] Detect the repeatable reference products R1, R2 and R3, each sample has 10 parallels for each concentration, and use three batches of kits for detection, calculate the difference between the kits within the batch and between batches, and test the results of R1 and R2 The intra-assay and inter-assay differences of the kit were less than 5%, and R3 was negative.

[0041] Interfering experiment:

[0042] Take the mixed serum and add interfering substances, including: conjugated bilirubin, free bilirubin, hemoglobin, ascorbic acid, glyceride, th...

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PUM

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Abstract

A chlamydiae pneumoniae IgM chemiluminescent immunoassay kit is disclosed. The kit includes magnetic micro-particles coated with a chlamydiae pneumoniae antigen, an acridinium ester labeled mouse anti-human IgM antibody, a chlamydiae pneumoniae IgM calibrated product, a pre-excitation liquid and an excitation liquid. The invention also discloses a preparing method of the kit. Compared with kits at present, the kit has advantages of simple and convenient operation, high sensitivity, a wide detection range, and the like.

Description

technical field [0001] The invention relates to the field of in vitro diagnostic immunodetection, and specifically, the invention provides a chemiluminescence immunodetection kit for Chlamydia pneumoniae IgM and a preparation method thereof. Background technique [0002] Chlamydia pneumoniae ( Chlamydia pneumoniae , Cpn) is a strictly eukaryotic intracellular parasitic prokaryotic microorganism identified in 1989, which has been renamed Chlamydia pneumoniae ( Chlamydophila pneumoniae , Cpn), together with Chlamydia psittaci, Chlamydia abortus, Chlamydia guinea pigs, Chlamydia cats, and Chlamydia mammals form the genus Chlamydia. The size of the CPn genome is about 1.2Mbp, consisting of 21 Pmp genes, a type III secreted virulence factor system, 3 serine / threonine protein kinases, and 2 phospholipase-D-like protein genes. [0003] Cpn infection is very common and distributed worldwide, mainly causing respiratory tract infections in humans, which is positively correlated wit...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/76G01N33/577
CPCG01N21/76G01N33/577
Inventor 叶国强徐向红夏福臻钱纯亘刘星
Owner SHENZHEN YHLO BIOTECH
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