Indirect ELISA (Enzyme-Linked Immuno Sorbent Assay) detection method of SARS-CoV-2S protein IgG (Immunoglobulin G)

Abstract

The invention relates to an indirect ELISA detection method of SARSCoV2S protein IgG (Immunoglobulin G), which comprises the following steps: adding 100 muL of COVID-19 spike protein diluted by adopting an ELISA coating solution into each well of an elisa plate, standing overnight at 4 DEG C, and cleaning with a PBST (Phosphate Buffer Solution); adding 250 muL of a 5% skim milk powder solution forsealing, diluting the serum to be detected with an ELISA antibody diluent as a primary antibody, adding 100 muL of the primary antibody into each well, and incubating at 37 DEG C for 1 h; using horseradish peroxide labeled anti-human IgG diluted by an ELISA antibody diluent as a secondary antibody, adding 100 muL of the secondary antibody into each well, performing incubation at 37 DEG C for 45 min, adding 100 [mu] L of a single-component TMB color developing solution into each well in a dark place, performing incubation at 37 DEG C for 5 min, adding 50 muL of a stop solution into each well,wherein the detection wavelength is 450 nm, the reference wavelength is 630 nm; and detecting the absorbance value OD<450 nm> of 450 nm by a microplate reader.

Description

technical field [0001] The invention belongs to the technical field of virus detection and relates to an indirect ELISA detection method for SARS-CoV-2 S protein IgG. Background technique [0002] The new member of the Coronaviridae family discovered in December 2019 and associated with severe pneumonia was originally called 2019 novel coronavirus (2019-nCoV) and is now called severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2 ), causing coronavirus disease 2019 (COVID-19). SARS-CoV-2 is highly homologous to another human coronavirus (severe acute respiratory syndrome coronavirus (SARS-CoV)). SARS-CoV-2 is transmitted by droplets and possibly by the fecal-oral route. The symptoms of COVID-19 cases are dry cough, fever, body aches and weakness, and diarrhea. The clinical manifestations of severe cases are high fever, dyspnea, and low blood oxygen levels. Most of the deaths are caused by respiratory failure and multi-organ damage caused by cytokine storms. Exhauste...

AI Technical Summary

Problems solved by technology

At present, several polymerase chain reaction (PCR)-based technologies to detect viral RNA are the main methods to determine the diagnosis of new coronavirus infection, but the positive rate of nucleic acid detection based on respiratory specimens is low, and a single nucleic acid test result may cause missed diagnosis of new coronavirus patients. It is necessary to develop other methods to supplement nucleic acid detection. At the same time, real-time fluorescent quantitative PCR (quantitative real-timePCR RT-qPCR) can only detect whether there is virus in the specimen, but it cannot reflect the humoral immune response of the case

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