Sensitive identification of mutations in genes related to the pathogenesis of cancer is a prerequisite for risk-stratified therapies. Next-generation sequencing (NGS) in lymphoma has revealed genetic heterogeneity which makes clinical translation challenging. We established a 454-based targeted resequencing platform for robust high-throughput sequencing from limited material of patients with lymphoma. Hotspot mutations in the most frequently mutated cancer consensus genes were amplified in a two-step multiplex-polymerase chain reation (PCR) which was optimized for homogeneous coverage of all regions of interest. We show that targeted resequencing based on NGS technologies allows highly sensitive detection of mutations and assessment of clone size. The application of this or similar techniques will help the development of genotype-specific treatment approaches in lymphoma.
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http://dx.doi.org/10.3109/10428194.2013.796053 | DOI Listing |
Viruses
December 2024
Foundation Plant Services, University of California-Davis, Davis, CA 95616, USA.
Among the cultivated crop species, the economically and culturally important grapevine plays host to the greatest number of distinctly characterized viruses. A critical component of the management and containment of these viral diseases in grapevine is both the identification of infected vines and the characterization of new pathogens. Next-generation high-throughput sequencing technologies, i.
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December 2024
Wadsworth Center, David Axelrod Institute, New York State Department of Health, Albany, NY 12208, USA.
A historical perspective of more than one hundred years of influenza surveillance in New York State demonstrates the progression from anecdotes and case counts to next-generation sequencing and electronic database management, greatly improving pandemic preparedness and response. Here, we determined if influenza virologic surveillance at the New York State public health laboratory (NYS PHL) tests sufficient specimen numbers within preferred confidence limits to assess situational awareness and detect novel viruses that pose a pandemic risk. To this end, we analyzed retrospective electronic data on laboratory test results for the influenza seasons 1997-1998 to 2021-2022 according to sample sizes recommended in the Influenza Virologic Surveillance Right Size Roadmap issued by the Association of Public Health Laboratories and Centers for Disease Control and Prevention.
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November 2024
College of Plant Protection, Nanjing Agricultural University, Nanjing 210095, China.
Rice is a crucial staple food for over half the global population, and viral infections pose significant threats to rice yields. This study focuses on the Rice Stripe Virus (RSV), which is known to drastically reduce rice productivity. We employed RNA-seq and ribosome profiling to analyze the transcriptional and translational responses of RSV-infected rice seedlings.
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November 2024
MRC/UVRI & LSHTM Uganda Research Unit, Entebbe 256, Uganda.
The emergence of SARS-CoV-2 variants has heightened concerns about vaccine efficacy, posing challenges in controlling the spread of COVID-19. As part of the COVID-19 Vaccine Effectiveness and Variants (COVVAR) study in Uganda, this study aimed to genotype and characterize SARS-CoV-2 variants in patients with COVID-19-like symptoms who tested positive on a real-time PCR. Amplicon deep sequencing was performed on 163 oropharyngeal/nasopharyngeal swabs collected from symptomatic patients.
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November 2024
Institute of Biology, ELTE Eötvös Loránd University, 1117 Budapest, Hungary.
The increasingly widespread application of next-generation sequencing (NGS) in clinical diagnostics and epidemiological research has generated a demand for robust, fast, automated, and user-friendly bioinformatics workflows. To guide the choice of tools for the assembly of full-length viral genomes from NGS datasets, we assessed the performance and applicability of four open-source bioinformatics pipelines (shiver-for which we created a user-friendly Dockerized version, referred to as dshiver; SmaltAlign; viral-ngs; and V-pipe) using both simulated and real-world HIV-1 paired-end short-read datasets and default settings. All four pipelines produced consensus genome assemblies with high quality metrics (genome fraction recovery, mismatch and indel rates, variant calling F1 scores) when the reference sequence used for assembly had high similarity to the analyzed sample.
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