Joint inspection reagent for encephalitis
A reagent and joint inspection technology, applied in the field of immunoassay, can solve the problems of reducing detection accuracy and false positives, and achieve the effects of saving detection time, strong specificity, and improving accuracy
- Summary
- Abstract
- Description
- Claims
- Application Information
AI Technical Summary
Problems solved by technology
Method used
Image
Examples
Embodiment 1
[0041] Example 1: Preparation of reagents for joint detection of encephalitis
[0042] A combined detection reagent for encephalitis, the reagent comprises a well plate reagent, a goat anti-human secondary antibody labeled with Dylight 488, and a phosphate buffer; wherein the well plate is a 96-well plate, and the well plate reagent comprises NMDAR, LGI1 expressing labeled mCherry fluorescent protein , GABARB1 / B2, AMPAR1, AMPAR2, CASPR2 transfected cells and blank vector expressing mCherry-labeled fluorescent protein transfected cells, transfected cells in different sample wells.
[0043]1) NMDAR, LGI1, GABARB1 / B2, AMPAR1, AMPAR2, CASPR2 transfected cells expressing marker mCherry (red fluorescent protein) and blank vector transfecting cells expressing marker mCherry were cultured in the sample wells of 96-well plate, respectively. Observe the cell growth under a microscope, and wait for the cells to rise to nearly 100%;
[0044] 2) Remove the medium and wash with 1×PBS;
[...
Embodiment 2
[0047] Example 2: Encephalitis joint detection reagent for the detection of cerebrospinal fluid
[0048] The reagents prepared in Example 1 are used for the detection of anti-NMDAR antibody, anti-LGI1 antibody, anti-GABARB1 / B2 antibody, anti-AMPAR1 antibody, anti-AMPAR2 antibody and anti-CASPR2 antibody as follows:
[0049] 1) Prepare cerebrospinal fluid samples, add 30µl-50µl of diluted cerebrospinal fluid to the sample wells on the 96-well plate;
[0050] 2) Incubate at 37°C for 1 hour or at 4°C overnight;
[0051] 3) Wash 3 times with 1×PBS, 3 minutes each time;
[0052] 4) Drain 1×PBS thoroughly, add 30µl-50µl fluorescent Dylight 488-labeled goat anti-human secondary antibody to the sample wells on the plate, and incubate at 37°C for 30 minutes;
[0053] 5) Wash 3 times with 1×PBS, 3 minutes each time;
[0054] 6) Add 30µl-50µl of 1×PBS to the sample well;
[0055] 7) Place the well plate under a fluorescence microscope for microscopic examination.
Embodiment 3
[0056] Example 3: Interpretation of anti-NMDAR antibodies in samples by fluorescence microscopy
[0057] The interpretation method of the orifice plate that embodiment 2 handles is as follows:
[0058] 1) Use a fluorescence microscope to observe the emitted fluorescent spots at a wavelength of 518 nm and 610 nm, respectively, by observing whether green fluorescent spots or red fluorescent spots are emitted;
[0059] 2) Compare the positions and features of the green fluorescent spots with the positions of the red fluorescent spots to see if they overlap;
[0060] 3) If the position of the green fluorescent spot and the position of the red fluorescent spot can overlap, and the characteristics are basically the same, it can be determined that the sample contains anti-NMDAR antibody;
[0061] 4) If the green fluorescent spots cannot be emitted; or the positions of the green fluorescent spots and the red fluorescent spots cannot overlap; or the positions of the green fluorescent ...
PUM
Login to View More Abstract
Description
Claims
Application Information
Login to View More