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MeSH Disease

HGNC Genes

SARS-CoV-2 proteins

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SARS-CoV-2 Proteins
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    Development and validation of cost-effective one-step multiplex RT-PCR assay for detecting the SARS-CoV-2 infection MESHD using SYBR Green melting curve analysis

    Authors: Shovon Lal Sarkar; A. S. M. Rubayet Ul Alam; Prosanto Kumar Das; Md. Hasan Ali Pramanik; Hassan M. Al-Emran; Iqbal Kabir Jahid; Md. Anwar Hossain

    doi:10.1101/2021.05.06.21256629 Date: 2021-05-08 Source: medRxiv

    TaqMan probe-based expensive commercial real-time (RT) PCR kits are being used in COVID-19 MESHD diagnosis. The unprecedented scale of SARS-CoV-2 infections MESHD has urgently needed to meet the challenge of testing more persons at a reasonable cost. This study developed a rapid, simple, and cost-effective alternative diagnostic method based on melting curve analysis of SYBR green multiplex assay with a host-specific internal control. A total of 90 randomly selected samples were used for comparing the assay with an available commercial kit to analyse the variation and validity of this in-house developed method. Our customized designed primers specifically detected the virus as similar to commercial kit manufactured by Sansure Biotech Inc. We optimized separately the N, E, S, and RdRp PROTEIN genes by SYBR Green RT-PCR method based on melting curve analysis. Afterwards, a multiplex COVID-19 MESHD diagnosis method targeting N and E genes PROTEIN of the virus along with the { beta}-actin HGNC gene of the host as an internal control has been established. The total run-time of our proposed method was less than 90 minutes. The cost of each sample processing was less than $2. Overall, this one-step and one-tube method can revolutionize the COVID-19 MESHD diagnosis in developing countries.

    A new system in qualitative RT-PCR detecting SARS-CoV-2 in biological samples: an Italian experience.

    Authors: Marco Favaro; Walter Mattina; Enrico Salvatore Pistoia; Roberta Gaziano; Paolo Di Francesco; Simon Middleton; Silvia D'Angelo; Tullio Altarozzi; Carla Fontana

    doi:10.1101/2020.06.17.20124396 Date: 2020-06-19 Source: medRxiv

    In the last moths the world was faced with the pandemic of a new severe acute respiratory syndrome coronavirus (SARS-CoV) MESHD and the majority of the Nations have yet to come out of it. Numerous assays have emerged to meet SARS-CoV-2 diagnostic needs. A clear knowledge of these assays parameters is essential to choose the proper test by clinical microbiologists. Unfortunately, the latter cannot be the unique criterion that guides test selection as - given the great demand - shortcomings of commercial kits is also a great issue. Aimed by the intention of overcoming both difficulties we have developed a new qualitative RT-PCR probe based for COVID-19 MESHD detection. The system detects three genes of SARS-CoV-2: RNA-dependent RNA polymerase PROTEIN ( RdRp PROTEIN), envelope (E) and nucleocapsid (N PROTEIN) and { beta}-actin HGNC gene used as endogenous internal control. The results of our assay show a total agreement with those obtained using a commercially available kit, with the exception of two specimens which did not pass the endogenous internal control. Moreover, our kit was designed to be open either for nucleic acid extraction step or on the RT-PCR assay to be carried out on several instruments. Thus, it is free from the industrial production logics of closed systems and conversely it is hypothetically available for distribution on large numbers in any microbiological laboratories. Presently, the kit is currently distributed worldwide

The ZB MED preprint Viewer preVIEW includes all COVID-19 related preprints from medRxiv and bioRxiv, from ChemRxiv, from ResearchSquare, from arXiv and from Preprints.org and is updated on a daily basis (7am CET/CEST).
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MeSH Disease
HGNC Genes
SARS-CoV-2 Proteins


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