Adsorbent for detecting IgM antibody and preparation method of adsorbent

A technology of antibody detection and adsorbent, applied in the field of adsorbent for IgM antibody detection and preparation thereof, can solve the problems of inaccurate results and high background of results, achieve simple reagents, good sensitivity and specificity, and improve detection accuracy. Effect

Inactive Publication Date: 2017-01-04
TAIZHOU ZECEN BIOTECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The technical problem to be solved by the present invention is to overcome the defect that the capture method or the indirect method is interfered by IgG to varying degree

Method used

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  • Adsorbent for detecting IgM antibody and preparation method of adsorbent
  • Adsorbent for detecting IgM antibody and preparation method of adsorbent
  • Adsorbent for detecting IgM antibody and preparation method of adsorbent

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0016] Embodiment 1 Toxoplasma gondii IgM (TOX IgM) antibody detection reagent (magnetic particle chemiluminescence method)

[0017] Source of samples: 5 cases were positive and 5 cases were negative for Toxoplasma gondii IgM antibody in clinical screening. Toxoplasma gondii IgG was purchased from manufacturer 2, and RF was purchased from manufacturer 3.

[0018] Comparison of different types and different concentrations of PEG:

[0019] Adsorbent A (PEG2000): 10 mM PB buffer, 150 mM NaCl, 6% polyethylene glycol 2000 (PEG2000).

[0020] Adsorbent B (PEG4000): 10 mM PB buffer, 150 mM NaCl, 6% polyethylene glycol 4000 (PEG4000).

[0021] Adsorbent C (PEG6000): 10 mM PB buffer, 150 mM NaCl, 6% polyethylene glycol 6000 (PEG6000).

[0022] Adsorbent D (PEG20000): 10 mM PB buffer, 150 mM NaCl, 6% polyethylene glycol 20000 (PEG20000).

[0023] TOX IgG and RF factors were added to negative samples, and different types of PEG were reacted with the same batch of reagents to detect TO...

Embodiment 2

[0059] Embodiment 2-cytomegalovirus IgM (CMV IgM) antibody detection reagent (magnetic particle chemiluminescence method)

[0060] Source of samples: Clinical screening of CMV IgM antibody was positive in 5 cases and negative in 5 cases. Giant cell IgG was purchased from manufacturer 2, and RF was purchased from manufacturer 3.

[0061] Detection steps:

[0062] (1) After diluting the sample 1:100, take 30ul of the diluted sample and mix it with 30ul of adsorbent. After mixing, react at 37°C for 5min.

[0063] (2) Add 30ul CMV-FITC, mix well, and react at 37°C for 15min.

[0064] (3) Add 30ul magnetic beads coupled with FITC antibody, mix well, and react at 37°C for 5min.

[0065] (4) Wash 3 times with 10mM PBS and pat dry.

[0066] (5) Add 60ul of AP-labeled anti-human IgM and react at 37°C for 15min.

[0067] (6) Wash 3 times with 10mM PBS and pat dry.

[0068] (7) Add substrate and read.

[0069] (8) The result is compared with the critical value to obtain the ratio, ...

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Abstract

The invention discloses an adsorbent for detecting an IgM antibody. A phosphate buffer solution of 10mM of PB and 150mM of NaCl and with the pH of 7.2 to 7.4 is taken as a base, polyethylene glycol 6000 of which the final concentration is 6wt%, 1mg/mL of an anti-human IgG antibody, 10mg/mL of BSA and 0.1wt% of proclin 300 are added, and the titer of the IgG antibody is not less than 1: 300000. The adsorbent is simple in reagent composition, and cheap in price of the anti-human IgG and PEG6000 which are taken as main reagents and mild in reaction, the pH of a buffer system of the adsorbent leans neutrality, and the ionic strength is suitable, thereby having little effect on the immune response of an antigen antibody; the adsorbent further has better sensitivity and specificity, a better absorption effect on specific IgG and RF factors and a good application range for most of IgM detection reagents, especially for indirect-method determination of IgM, and can obviously lower sample backgrounds which affect detected IgM antibody results and improve the detection accuracy.

Description

technical field [0001] The invention relates to the field of antibody detection, in particular to an adsorbent for IgM antibody detection and a preparation method thereof. Background technique [0002] IgM antibody is the earliest specific antibody in humoral immunity. When pathogens infect the human body, the detection of IgM antibody is of great significance for the early diagnosis of infectious diseases. Currently, the most widely used methods on the market include capture method and indirect method, which have been applied to detection platforms such as Elisa, colloidal gold, and chemiluminescence. When the capture method is used to measure IgM, some clinical samples contain rheumatoid factor against self-denatured IgG in different concentrations, and there are IgM antibodies in them, which will react with anti-human IgM antibodies and cause false positives. The indirect method for IgM determination has a more obvious impact, because specific IgM antibodies against cert...

Claims

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Application Information

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IPC IPC(8): G01N33/68C07K16/00C07K1/22
CPCG01N33/6854C07K16/00
Inventor 王振刘振国陈海生鲁衡
Owner TAIZHOU ZECEN BIOTECH CO LTD
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