Module- Human haemotology and clinical immunology.make a lab report on topic - Covid - 19 Total antibody ELISA.

Title: Covid - 19 Total antibody ELISA
Abstract    2
Introduction    2
Method    3
Discussion    4
Conclusion    5
Reference    7
Abstract
Over 2 million people have died due to COVID-19, brought on by the SARS-CoV-2 virus and has affected people's daily lives worldwide. Nucleic acid PCR assays, direct viral antigen testing, and indirect human antibodies tests particular to SARS-CoV-2 are being used as mainstay diagnostic techniques in laboratories to identify active infections and past exposure (Al-Jighefee et al,2021). In this viewpoint, we briefly discuss the PCR and antigenic tests before concentrating mainly on current blood work and their shortcomings, such as errors and potential unreliability. False-negative results in monoclonal immunoassays can result from the assay design, choice of intracellular pathogens and antibody types, molecular diagnostic windows, individual variability, and changes in antibody levels. The most common causes of wrongful convictions in antibody immunoassays are autoimmune illness and antibody cross-reactivity with other viruses. False positives and false negatives are also impacted by spectrum bias (Ejazi, Ghosh and Ali,2021). To increase sensitivity and specificity, assay makers must carefully evaluate the choice of intracellular pathogens and composition of human antibodies in addition to improving test designs. To test patients utilising currently flawed but accessible tests with intelligent strategies and reasonable assessment of the test results, doctors must consider the variables impacting the accuracy of assays.
Introduction
The coronavirus illness 2019, initially identified in Wuhan, China, in December 2019, soon spread over the globe to cause a pandemic. The World Health Organization (WHO) classified the outbreak as a pandemic on January 30th of, 2020 and March 11th of the same year, respectively (Al-Jighefee et al,2021). More than 100 million COVID-19 incidents have been reported, according to the Center for Systems Engineering and Sciences at Johns Hopkins University have been documented as of 27 January 2021 in more than 188 nations and territories, culminating in more than 2 million fatalities.
In attempts to confine populations, population quarantine (or "lock-down") measures have been frequently utilised to restrict movement and reduce interpersonal interaction. Scaling up diagnostic testing is urgently required, along with comprehensive data collection on current and past SARS-CoV-2 exposure at the individual and community levels. Diagnostic testing must include mass screening as well as a screening of particular high-risk categories (contact information of confirmed cases, healthcare workers, and their relatives). The International Committee on Taxonomy of Viruses named this virus SARS coronavirus ("SARSCoV2") on February 11, 2020, due to its genetic resemblance to the coronavirus that caused the (2003) severe acute respiratory syndrome (SARS) outbreak. The S1 subunit has a receptor-binding domain (RBD), which facilitates coronavirus entry into host cells. The N-terminal domain (NTD) and also the C-terminal domain (CTD), which are considered to engage with viral RNA genome most likely through electrostatic interactions, are two different RNA-binding domains on the dimeric N protein. In this perspective, with the help of multiple studies, researchers will provide a brief overview of the primary PCR and antigen tests used to identify COVID-19 disease activity (Thomas et al,2021). The numerous press reports of errors in antibody assays prompt us to shift our attention to antibody assays and arrays subsequently. Examining the flaws in the way antibody testing is currently done, including false negatives and positives as well as the various possible causes of inaccuracy.
The creation of a new ELISA targets the SARS-CoV spike protein. Using SARS2 Spike that has been mammalian codon optimised and has had a GSAS substitution made at the furin cleavage site and a double proline substitution made at amino acids 986 and 987, recombinant SARS-CoV-2 trimeric spiked protein was created as stated (Wan, Seng and Lim,2020). The T4 fibrin pattern, HRV3C protease cleavage location, a TwinStrep Tag, and an 8-HisTag were placed after the C-terminal. The gene was expressed in 293T cells after being cloned into a pHLsec. The transgenic S protein was purified using the HIS trap HP column (Liu and Rusling,2021).
To find S protein antibodies, we employed ELISA. StrepMAB-Classic was applied as a coating on MAXISORP Immuno plates. 0.125 g of...