https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9581791

Evidence for SARS-CoV-2 Delta and Omicron co-infections and recombination

Abstract Between November 2021 and February 2022, SARS-CoV-2 Delta and Omicron variants co-circulated in the United States, allowing for co-infections and possible recombination events. We sequenced 29,719 positive samples during this period and analyzed the presence and fraction of reads supporting mutations specific to either the Delta or Omicron variant. We identified 18 co-infections, one of which displayed evidence of a low Delta-Omicron recombinant viral population. We also identified two independent cases of infection by a Delta-Omicron recombinant virus, where 100% of the viral RNA came from one clonal recombinant. In the three cases, the 5’-end of the viral genome was from the Delta genome, and the 3’-end from Omicron including the majority of the spike protein gene, though the breakpoints were different. Delta-Omicron recombinant viruses were rare, and there is currently no evidence that Delta-Omicron recombinant viruses are more transmissible between hosts compared to the circulating Omicron lineages. 

/pdf/evidence-for-sars-cov-2-delta-and-omicron-co-infections-and-3h0pn20v.pdf

The continued circulation of SARS-CoV-2 amid limited surveillance efforts and inconsistent vaccination of populations has resulted in the emergence of variants that uniquely impact public health systems. Thus, in conjunction with functional and clinical studies, continuous detection and identification are quintessential to informing diagnostic and public health measures. ABSTRACT Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) variants are characterized by differences in transmissibility and response to therapeutics. Therefore, discriminating among them is vital for surveillance, infection prevention, and patient care. While whole-genome sequencing (WGS) is the “gold standard” for variant identification, molecular variant panels have become increasingly available. Most, however, are based on limited targets and have not undergone comprehensive evaluation. We assessed the diagnostic performance of the highly multiplexed Agena MassARRAY SARS-CoV-2 Variant Panel v3 to identify variants in a diverse set of 391 SARS-CoV-2 clinical RNA specimens collected across our health systems in New York City, USA and Bogotá, Colombia (September 2, 2020 to March 2, 2022). We demonstrated almost perfect levels of interrater agreement between this assay and WGS for 9 of 11 variant calls (κ ≥ 0.856) and 25 of 30 targets (κ ≥ 0.820) tested on the panel. The assay had a high diagnostic sensitivity (≥93.67%) for contemporary variants (e.g., Iota, Alpha, Delta, and Omicron [BA.1 sublineage]) and a high diagnostic specificity for all 11 variants (≥96.15%) and all 30 targets (≥94.34%) tested. Moreover, we highlighted distinct target patterns that could be utilized to identify variants not yet defined on the panel, including the Omicron BA.2 and other sublineages. These findings exemplified the power of highly multiplexed diagnostic panels to accurately call variants and the potential for target result signatures to elucidate new ones. IMPORTANCE The continued circulation of SARS-CoV-2 amid limited surveillance efforts and inconsistent vaccination of populations has resulted in the emergence of variants that uniquely impact public health systems. Thus, in conjunction with functional and clinical studies, continuous detection and identification are quintessential to informing diagnostic and public health measures. Furthermore, until WGS becomes more accessible in the clinical microbiology laboratory, the ideal assay for identifying variants must be robust, provide high resolution, and be adaptable to the evolving nature of viruses like SARS-CoV-2. Here, we highlighted the diagnostic capabilities of a highly multiplexed commercial assay to identify diverse SARS-CoV-2 lineages that circulated from September 2, 2020 to March 2, 2022 among patients seeking care in our health systems. This assay demonstrated variant-specific signatures of nucleotide/amino acid polymorphisms and underscored its utility for the detection of contemporary and emerging SARS-CoV-2 variants of concern.

https://doi.org/10.1101/2022.05.28.22275691

A Robust, Highly Multiplexed Mass Spectrometry Assay to Identify SARS-CoV-2 Variants

Emergence of new variants of Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) during current pandemic of Coronavirus Disease 2019 (COVID-19) and several waves of infections by some of variants emphasized the importance of continuous surveillance. While genomic surveillance through whole genome sequencing is performed as a standard method, identification of known variants through mutation-targeting molecular diagnosis such as qRT-PCR is also useful for timely investigation. However, there are limited studies regarding the concurrent detection and identification of SARS-CoV-2 variants through a LAMP-based method. In this study, we developed and evaluated RT-LAMP assays to detect characteristic deletions of SARS-CoV-2 variants. In addition, we evaluated a fluorescent probe mediated method for identification of single nucleotide substitution by RT-LAMP. Finally, we discussed restrictions and perspectives regarding pathogen screening and surveillance of variants by RT-LAMP based on our observations.

Development and Evaluation of RT-LAMP Assays to Identify Variants of SARS-CoV-2

Foz do Iguassu is a famous Brazilian tourist site located on the Triple Frontier, where Brazil has borders with Argentina and Paraguay, and it is home of the world second largest hydroelectric power plant, named Itaipu Binational. SARS-CoV-2 wastewater monitoring is an important tool that has been helping health authorities to deal and plan the fight against COVID-19 ongoing pandemic. Since 2020, Foz do Iguassu has been weekly monitoring 20 points in the city and at the Itaipu Hydroelectric Power Plant. Samples from January/2022 were sequenced using Nanopore Technology and MinION device. All viable samples resulted in the presence of Omicron variant. The samples presented three unique mutations (T13195C ORF1ab, G14966- ORF1ab and A27259C ORF6) that are not defining mutations from any of the Variants of Concern (VOCs). One of these mutations (T13195C ORF1ab) is described as a signature mutation found in Omicron sequences from patients of two hospitals in Washington, United States. Since Foz do Iguassu receives thousands of visitors from all around the world, we suggest that the current Omicron variant circulating in the city could be originated from United States. We demonstrated that, despite inherent difficulties associated with sequencing environmental samples, sequencing SARS-CoV-2 using Nanopore technologies as a complement of the wastewater surveillance is feasible and can help health authorities to make improvements and determinations to act against the COVID-19 pandemic.

First Detection of SARS-CoV-2 Omicron Variant Using Nanopore Sequencing in Wastewater of Foz Do Iguassu-Brazil

The rapid emergence of new SARS-CoV-2 variants raises a number of public health questions including the capability of diagnostic tests to detect new strains, the efficacy of vaccines, and how to map the geographical distribution of variants to better understand patterns of transmission and possible load on healthcare resources. Next-Generation Sequencing (NGS) is the primary method for detecting and tracing the emergence of new variants, but it is expensive, and it can take weeks before sequence data is available in public repositories. Here, we describe a Polymerase Chain Reaction (PCR)-based genotyping approach that is significantly less expensive, accelerates reporting on SARS-CoV-2 variants, and can be implemented in any testing lab performing PCR. Specific Single Nucleotide Polymorphisms (SNPs) and indels are identified that have high positive percent agreement (PPA) and negative percent agreement (NPA) compared to NGS for the major genotypes that circulated in 2021. Using a 48-marker panel, testing on 1,128 retrospective samples yielded a PPA and NPA in the 96.3 to 100% and 99.2 to 100% range, respectively, for the top 10 most prevalent lineages. The effect on PPA and NPA of reducing the number of panel markers was also explored. In addition, with the emergence of Omicron, we also developed an Omicron genotyping panel that distinguishes the Delta and Omicron variants using four (4) highly specific SNPs. Data from testing demonstrates the capability to use the panel to rapidly track the growing prevalence of the Omicron variant in the United States in December 2021.

A method for variant agnostic detection of SARS-CoV-2, rapid monitoring of circulating variants, detection of mutations of biological significance, and early detection of emergent variants such as Omicron

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A method for variant agnostic detection of SARS-CoV-2, rapid monitoring of circulating variants, detection of mutations of biological significance, and early detection of emergent variants such as Omicron