Autoclave the solution and store at room temperature. Mix for about 15 min and adjust pH to 7.4 with HCl. Phosphate-buffered saline (PBS 10×): Weigh and transfer 80 g NaCl, 2 g KCl, 14.4 g Na 2HPO 4, and 2.4 g KH 2PO 4 to a 1 L graduated cylinder, and add about 800 mL water. MaxiSorp flat-bottom 96-well ELISA plates. Diligently follow all waste disposal regulations when disposing waste materials. Prepare all solutions using deionized water and store them at room temperature or 4 ☌. This assay was developed and validated using a large number of well-characterized human serum samples and could be adapted by any laboratory especially that all required reagents are commercially available. Here, we provide a detailed protocol for indirect ELISA based on recombinant MERS-CoV S1 subunit (amino acids 1–725) for qualitative and quantitative MERS-CoV serological testing. The importance of validated ELISA is evident especially when dealing with pathogens requiring high containment facilities such as MERS-CoV as it could provide a rapid, simple, and cheap method for field or clinical use without the need for high containment laboratories.ĭifferent indirect ELISAs based on MERS-CoV nucleocapsid (N) or spike (S) proteins were developed and used in epidemiological and surveillance studies.
The use of an enzyme-linked conjugate as well as substrate leads to changes in color which help to determine the presence and the quantity of substances such as peptides, proteins, antibodies, and hormones in a given sample, such as blood or urine samples. Immunosorbent assay (ELISA) is an immunological technique used to study the interaction between antigens and antibodies.
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