Parainfluenza virus type-1, 2, 3 chemiluminescence immunoassay kit and preparation method thereof

A technology of chemiluminescent immunity and detection kits, applied in chemiluminescence/bioluminescence, analysis by making materials react chemically, measuring devices, etc., can solve cross-contamination of reagents, cannot perform single-serve detection and filling Reagent operation is cumbersome and other problems, to achieve the effect of high detection accuracy

Inactive Publication Date: 2016-10-26
SHENZHEN YHLO BIOTECH
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  • Abstract
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AI Technical Summary

Problems solved by technology

[0007] (1) Use 12×8 type, 6×8 type, 8×12 type or whole plate type 96-well special microwell plate as antigen coating equipment and reaction container, which can only be divided into 12 batches and 6 batches when used , 8 batches or the whole board can be used at one time, and independent and single-person testing cannot be carried out;
[0008] (2) There are many types of reagents used in quantitative determination, and each detection reagent must be contained in a reagent bottle, and each time a reagent is used, it is necessary to replace the suction nozzle to fill it into the microwells of the microwell plate , not only there are many types of reagent bottles, but also the operation of filling reagents is extremely cumbersome;
[0009] (3) There is a lack of corresponding labeling of the testing information. The production batch number and expiry date information of the testing reagent can only be known or known by checking the label on the outer packaging box of the kit, and the known information is not controlled during the testing process, which has great potential. large randomness;
[0010] (4) The detection reagents are in an open space during the detection process, which may easily cause cross-contamination among various reagents and affect the accuracy of the detection results;
[0011] (5) Manual operation is mostly used in the detection process, the addition of reagents or samples is not very precise, the operation process is extremely cumbersome and complicated, and operation errors are prone to occur, and the accuracy and precision of the detection results are poor;
[0012] (6) The quantity configuration and use of the complete set of reagents for the test items are the number of items × 48 / 96 persons. If 10 items need to be tested, the configuration and use of reagents must be 10×48 / 96 persons. If Only one sample needs to detect 10 different items, and it also needs to configure reagents for 10×48 / 96 people, which has the disadvantage of not being economical and reasonable
[0017] From the existing detection methods of parainfluenza virus types 1, 2, and 3, it can be seen that although EIA, virus isolation, and RT-PCR diagnostic methods have the advantages of certain specificity and sensitivity, they require professional and technical personnel, Disadvantages of specialized instruments and equipment, specific conditions and time-consuming

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  • Parainfluenza virus type-1, 2, 3 chemiluminescence immunoassay kit and preparation method thereof
  • Parainfluenza virus type-1, 2, 3 chemiluminescence immunoassay kit and preparation method thereof
  • Parainfluenza virus type-1, 2, 3 chemiluminescence immunoassay kit and preparation method thereof

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preparation example Construction

[0055] Such as figure 1 The preparation method of the above-mentioned parainfluenza virus type 1, 2, and 3 chemiluminescence immunoassay kits shown comprises the following steps:

[0056] Take the suspension of carboxylated magnetic particles, remove the supernatant by magnetic separation, resuspend with MES buffer, then add EDC aqueous solution to activate the carboxyl groups on the surface of carboxylated magnetic particles, and then add parainfluenza virus type 1, 2, and 3 for recombination The protein was suspended at room temperature for 2h-10h, and the supernatant was removed by magnetic separation, and then resuspended with Tris buffer to obtain carboxylated magnetic particles coated with parainfluenza virus type 1, 2, and 3 recombinant proteins.

[0057] MES (2-(N-morpholine)ethanesulfonic acid) buffer had a concentration of 0.02M and a pH of 5.5.

[0058] Tris buffer has a concentration of 0.1 M and contains 2% BSA, pH 8.0.

[0059] The concentration of EDC (1-ethyl...

Embodiment 1

[0075] Example 1: Preparation of parainfluenza virus type 1, 2, and 3 chemiluminescent immunoassay kits

[0076] (1) Preparation of carboxylated magnetic particles coated with parainfluenza virus type 1, 2, and 3 recombinant proteins:

[0077] Take the suspension containing 50 mg of carboxylated magnetic particles (MagnaBind21353) with a particle size of 0.05 μm~1 μm, magnetically separate to remove the supernatant, resuspend with 0.02 M, pH 5.5 MES buffer, add 1 mL of newly prepared 10 mg / mL EDC aqueous solution, activate the carboxyl groups on the surface of magnetic beads, add 4 mg of parainfluenza virus type 1, 2, and 3 recombinant proteins (biorbyt, product number orb48780), suspend at room temperature for 6 hours, magnetically separate, remove the supernatant, and wash with 0.1M containing 2% BSA , resuspended to 1 mg / mL in Tris buffer with a pH of 8.0 to obtain carboxylated magnetic particles coated with recombinant proteins of parainfluenza virus types 1, 2, and 3, and...

Embodiment 2

[0082] Embodiment 2: Parainfluenza virus type 1, 2, 3 chemiluminescent immunoassay method

[0083] The automatic chemiluminescent immunoassay analyzer (YHLO, catalog number iFlash3000) was used as the detection tool, and the methodological mode was indirect immunoassay, that is, 50 μL of samples and 50 μL of parainfluenza virus type 1, 2, and 3 recombinant protein packages were sequentially added to the instrument. Carboxylated magnetic particles and 50 μL of parainfluenza virus type 1, 2, and 3 treatment solutions were reacted for 20 minutes, and then 50 μL of anti-human immunoglobulin acridinium ester was added for 20 minutes of reaction before magnetic separation. , the instrument sends the reaction mixture into the dark room, and sequentially adds the luminescent substrate A solution (H 2 o 2 ) and solution B (NaOH) for luminescence reaction, and finally record the luminescence value.

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Abstract

The invention discloses a parainfluenza virus type-1, 2, 3 chemiluminescence immunoassay kit and a preparation method thereof. The parainfluenza virus type-1, 2, 3 chemiluminescence immunoassay kit comprises carboxylic magnetic particles coated with parainfluenza virus type-1, 2, 3 recombinant proteins and chemiluminescent labels marked by anti-human immune globulin. The parainfluenza virus type-1, 2, 3 chemiluminescence immunoassay kit can adopt a full-automatic chemiluminescence immunoassay analyzer as a detection tool to complete the detection of the parainfluenza viruses type-1, 2, 3. By virtue of experiments, the detection sensitivity of the parainfluenza virus 1, 2, 3 chemiluminescence immunoassay kit reaches 1U / L, which is at least increased by 10 times compared with the sensitivity of the traditional detection method for the parainfluenza viruses type-1, 2, 3. The detection precision of the parainfluenza virus type-1, 2, 3 chemiluminescence immunoassay kit is relatively high.

Description

technical field [0001] The invention relates to the field of in vitro detection, in particular to a parainfluenza virus type 1, 2, 3 chemiluminescence immunoassay kit and a preparation method thereof. Background technique [0002] Human Parainfluenzavirus (HPIV) is a paramyxovirus [0003] Single-celled RNA viruses can be divided into four serotypes according to their antigenicity and heredity: human parainfluenza virus type 1 (HPIV1), human parainfluenza virus type 2 (HPIV2), and human parainfluenza virus type 3 ( HPIV3) and human parainfluenza virus type 4 (HPIV4). Human parainfluenza virus is a common pathogen of acute viral respiratory tract infection. It mainly causes lower respiratory tract infection in infants and upper respiratory tract infection in adults. In addition, human parainfluenza virus has also been reported to infect the urinary tract, which is considered to be a cause of prostatitis. Influenza B virus is a common influenza epidemic virus, which is only...

Claims

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Application Information

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IPC IPC(8): G01N21/76G01N33/569
CPCG01N21/76G01N33/56983G01N2333/115
Inventor 夏福臻祝亮王刚钱纯亘黄湘东
Owner SHENZHEN YHLO BIOTECH
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