Whole-genome sequencing & mutational analysis of SARS-CoV-2 from patients' faecal samples reveal the possible role in faecal-oral transmission

Rapid Detection of Predominant SARS-CoV-2 Variants Using Multiplex High-Resolution Melting Analysis

“…First, in terms of the detection reagent, the multiplex HRM method requires only a single common primer pair for target mutation site testing without the need for a fluorescent probe. Moreover, a number of studies have indicated that EvaGreen dye is sensitive, reliable, and cost-effective ( 17 , 31 , 32 ), which is why EvaGreen was used in this study. Second, with respect to experimental design, the multiplex HRM method confers great flexibility and a low-cost alternative for analyzing viral genomes with high mutation rates.…”

“…HRM is an easy and reliable closed-tube analysis that has been applied for the identification of mutations by scanning or molecular typing in several research fields, such as epidemiology and microbiology. Exploiting the advantages of HRM, our team has used this approach for the detection of several pathogens and their mutation sites, for example, in Neisseria gonorrhoeae , Chlamydia trachomatis , Mycoplasma genitalium , and SARS-CoV-2 ( 16 – 19 ). In these works, we developed a one-step multiplex HRM method for SARS-CoV-2 detection and mutation site identification, consisting of five assays: one assay detected SARS-CoV-2, and the other four assays obtained 12 mutation sites for variant analysis ( 19 ).…”

Lack of evidence of viability and infectivity of SARS-CoV-2 in the fecal specimens of COVID-19 patients

“…The data obtained was evaluated using the entire genome of the SARS-CoV-2 Wuhan-Hu-1 isolate after sequencing (GenBank accession number MN908947.3). Five specimens with >90% coverage yielded consensus of the whole genome data of SARS-CoV-2 and the remaining five were with partial genomes (mean genome length: 60.5%; The successful isolation of multiple strains of SARS-CoV-2 has been performed using Vero CCL-81 cell lines at Indian Council of Medical Research-National Institute of Virology (ICMR-NIV), Pune City, Maharashtra state, India (20). We attempted isolation of SARS-CoV-2 from fecal specimens in Vero CCL-81 cell lines (21).…”

“…The successful isolation of multiple strains of SARS-CoV-2 has been performed using Vero CCL-81 cell lines at Indian Council of Medical Research- National Institute of Virology (ICMR-NIV), Pune City, Maharashtra state, India ( 20 ). We attempted isolation of SARS-CoV-2 from fecal specimens in Vero CCL-81 cell lines ( 21 ).…”