Background: The global COVID-19 pandemic has led to an urgent need for scalable methods for clinical diagnostics and viral tracking. Next generation sequencing technologies have enabled large-scale genomic surveillance of SARS-CoV-2 as thousands of isolates are being sequenced around the world and deposited in public data repositories. A number of methods using both short- and long-read technologies are currently being applied for SARS-CoV-2 sequencing, including amplicon approaches, metagenomic methods, and sequence capture or enrichment methods. Given the small genome size, the ability to sequence SARS-CoV-2 at scale is limited by the cost and labor associated with making sequencing libraries.
Results: Here we describe a low-cost, streamlined, all amplicon-based method for sequencing SARS-CoV-2, which bypasses costly and time-consuming library preparation steps. We benchmark this tailed amplicon method against both the ARTIC amplicon protocol and sequence capture approaches and show that an optimized tailed amplicon approach achieves comparable amplicon balance, coverage metrics, and variant calls to the ARTIC v3 approach.
Conclusions: The tailed amplicon method we describe represents a cost-effective and highly scalable method for SARS-CoV-2 sequencing.
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http://dx.doi.org/10.1186/s12864-020-07283-6 | DOI Listing |
Int J Mol Sci
December 2024
College of Life Sciences, North China University of Science and Technology, Tangshan 063200, China.
Due to the price and demand of having increased dramatically, adulteration with other fungi is a common problem. Thus, a reliable method of authentic identification is essential. In the present work, a rapid DNA extraction and double-tailed recombinase polymerase amplification (RPA) coupled with nucleic acid hybridization lateral flow strip (NAH-LFS) was developed to distinguish authentic ingredients from other fungi substitutes.
View Article and Find Full Text PDFBiosensors (Basel)
December 2024
Department of Bioscience and Biotechnology, Konkuk University, 120 Neungdong-ro, Gwangjin-gu, Seoul 05029, Republic of Korea.
We developed a rapid and sensitive diagnostic platform that integrates isothermal viral gene amplification with a nucleic acid lateral flow assay (NALFA) to detect SARS-CoV-2 RNA. Isothermal gene amplification was performed by combining reverse transcription of viral RNA with recombinase polymerase amplification (RPA). In our diagnostic platform, DNA primers for the RPA reaction were modified by appending DNA tails, enabling the synthesis of tailed amplicon DNAs.
View Article and Find Full Text PDFHeliyon
October 2024
Universität Innsbruck, Department of Microbiology, Technikerstrasse 25d, 6020, Innsbruck, Austria.
Cofactor F is an electron carrier playing a crucial role in a variety of microorganisms during redox reactions of the primary and secondary metabolism due to its low redox potential and thus arouses increasing interest. In this study, cofactor F glutamyl tail length spectra in various habitats like manure, compost, soil, and digester sludge samples and their respective microbial communities were investigated using high performance liquid chromatography and an amplicon sequencing approach A previous study was used to identify F producing microorganisms. The highest concentration of cofactor F could be achieved in the horse manure, digester sludge, and mixed manure samples, which was approximately 100-fold higher than in all the other samples.
View Article and Find Full Text PDFPathogens
September 2024
Laboratory of Virology, Beijing Key Laboratory of Etiology of Viral Diseases in Children, Capital Institute of Pediatrics, Beijing 100020, China.
Head-to-tail sequences have been reported in human bocavirus (HBoV) 1-4. To reveal their features and functions, HBoV DNA was screened among respiratory specimens from pediatric patients with an acute respiratory infection (ARI) between April 2020 and December 2022, followed by HBoV genotyping. Head-to-tail sequences were detected using nested PCR, TA cloning, and Sanger sequencing, and these findings were confirmed by mNGS and amplicon sequencing.
View Article and Find Full Text PDFPlant Dis
September 2024
Henan Normal University, College of Life Science, Xinxiang, China;
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