SARS-CoV-2 IgM Antibody ELISA Kit

Principle of the Assay

 The kit uses enzyme-linked immunoassay. According to the structural characteristics of IgM antibodies, the same protein antigen was selected to coat the microporous plate and to be labeled with horseradish peroxidase (HRP); Labeled SARS-CoV-2-n recombinant protein (labeled antigen) with horseradish peroxidase (HRP). The kit design is as follows:

The 96-well microporous plate is coated with a recombinant SARS-CoV-2-n protein (solid-phase antigen); When the positive sample is to be tested (IgM) and the labeled antigen is added to the microplate, the pattern and product of (solid antigen +IgMAb+ labeled antigen) → (solid antigen -IgMAb-4 labeled antigen) will be formed. Absorbance was measured at 450nm wavelength by a microplate reader. The concentration of IgM antibody in the sample is directly proportional to the absorbance (OD value).

Target Information

 The new coronavirus belongs to the beta coronavirus of the genus β, which has an envelope, the particles are round or oval, often polymorphic, and the diameter is 60-140nm. Its genetic characteristics are significantly different from SARS-CoV and MERS-CoV. Current research shows that it has more than 85% homology with bat SARS-like coronavirus (bat-SLCOVZC45).

In vitro isolation and culture, SARS-CoV-2  can be found in human respiratory epithelial cells in about 96 hours, while it takes about 6 days to isolate and culture in Vero E6 and Huh-7 cell lines. Based on current epidemiological investigations, the incubation period is generally 7 days, with a maximum of 14 days. The main symptoms are fever, fatigue, and dry cough. A few patients have symptoms such as nasal congestion, runny nose, and diarrhea. In severe cases, dyspnea occurs more than a week later. In other severe cases, acute respiratory distress syndrome, septic shock, difficult to correct metabolic acidosis, and coagulation dysfunction develop rapidly. It is worth noting that in the course of severe and critically ill patients, there may be moderate to low fever, even without obvious fever. Some patients showed only low fever, mild fatigue, and no pneumonia and recovered after 1 week. In the early stages of the disease, the total number of white blood cells in the peripheral blood was normal or decreased, the lymphocyte count decreased, and some patients had increased liver enzymes, muscle enzymes, and myoglobin. Most patients have elevated C-reactive protein (CRP) and erythrocyte sedimentation rate and normal procalcitonin. In severe cases, D-dimer increases and peripheral blood lymphocytes progressively decrease. New coronavirus nucleic acids can be detected in throat swabs, sputum, lower respiratory tract secretions, and blood.

 

Materials Supplied

Kit Components 96 Wells Quantity/Size
Microporous plate coated with SARS-CoV-2-N recombinant protein antigen. (8×12)
1 plate
IgM antibody negative reference: specification : 0.8ml/ bottle
1 bottle
IgM antibody positive reference: specification : 0.8ml/ bottle
1 bottle
Enzyme conjugate: SARS-CoV-2-n antigen with HRP labeled:  specification: 10.5ml/ bottle.
1 bottle
Color rendering solution A, the main component is H2O2.  specification: 5.0ml/ bottle.
1 bottle
Color rendering solution B, the main component is TMB.  specification: 5.0ml/ bottle.
1 bottle
Termination solution, mainly containing dilute H2SO4
specification : 5.0ml/ bottle.
1 bottle
Concentrated washing solution, 10 times concentrated washing solution, dilute before use. Specification: 50.0ml/ bottle.
1 bottle
Adhesive Films 4 pieces
Operating Instructions
1 copy

Storage

Storage Store at 2 - 8°C

Performance Characteristics

REPEATABILITY Tested the positive reference products of the enterprise. Repeated detection 10 times, all the results were positive.
SPECIFICITY After the destruction test of 2-mercapto ethanol in this kit; it is verified that the detected antibody is IgM type antibody.