Category: Vector & Virus
RT-qPCR Diagnostics: The “Drosten” SARS-CoV-2 Assay Paradigm
The reverse transcription quantitative polymerase chain reaction (RT-qPCR) is an established tool for the diagnosis of RNA pathogens. Its potential for automation has caused it to be used as a presence/absence diagnostic tool even when RNA quantification is not required. This technology has been pushed to the forefront of public awareness by the COVID-19 pandemic,…
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Pooled RT-qPCR testing for SARS-CoV-2 surveillance in schools – a cluster randomised trial
Background: The extent to which youngsters and adolescents contribute to SARS-CoV-2 transmission stays not absolutely understood. Novel high-capacity testing strategies might present real-time epidemiological knowledge in academic settings serving to to determine a rational method to forestall and decrease SARS-CoV-2 transmission. We investigated whether or not pooling of samples for SARS-CoV-2 detection by RT-qPCR is a…
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Development of a qPCR-based method for counting overwintering spruce budworm (Choristoneura fumiferana) larvae collected during fall surveys and for assessing their natural enemy load: a proof-of-concept study
Background: In japanese Canada, surveys of overwintering second-instar spruce budworm (Choristoneura fumiferana) larvae (‘L2s’) are carried out every fall to information insecticide utility choices within the following spring. These surveys contain the gathering of fir and spruce branches in chosen stands, adopted by the mechanical/chemical removing of larvae. The latter are then counted manually on filter…
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Improving sustainable hydrogen production from green waste: [FeFe]-hydrogenases quantitative gene expression RT-qPCR analysis in presence of autochthonous consortia
Background: Bio-hydrogen manufacturing through darkish fermentation of low-value waste is a potent and easy imply of recovering vitality, maximising the harvesting of decreasing equivalents to provide the cleanest gasoline amongst renewables. Following a number of place papers from firms and public our bodies, the hydrogen financial system is regaining curiosity, particularly together with round financial system…
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What’s the norm in normalization? A frightening note on the use of RT-qPCR in the livestock science
Reverse-Transcription quantitative PCR (RT-qPCR) offers a precious device to check gene expression with beautiful sensitivity. To retain its inferential energy, user-introduced technical variability have to be decreased and accounted for. Deciding on a set of stably expressed inside management genes (ICG), validated for every experimental situation/pattern set, is broadly accepted as a dependable approach to…
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Development of real-time RT-qPCR assays for the typing of two novel bluetongue virus genotypes derived from sheeppox vaccine
Beforehand, we reported the detection of two novel bluetongue virus (BTV) strains (SPvvvv/02 and SPvvvv/03), presumably representing new BTV genotypes, in a batch of sheeppox vaccine. We developed type-specific RT-qPCR assays (concentrating on genome section 2) for these two new BTV strains. The restrict of detection of each assays was 10 genome copies/μl and no…
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Identification and validation of reference genes for RT-qPCR analysis in fetal rat pancreas
The selection of reference gene is essential for quantitative reverse transcriptase-polymerase chain response (RT-qPCR) assay. To display screen and decide the appropriate reference genes in fetal rat pancreas, we chosen eight candidate reference genes (Gapdh, Actb, Rn18 s, B2m, Rpl13a, Tbp, Ywhaz and Ubc), and evaluated the fidelity of gene expression from fetal rat pancreases…
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An optimized stepwise algorithm combining rapid antigen and RT-qPCR for screening of COVID-19 patients
Background & intention: We investigated the mixture of speedy antigen detection (RAD) and RT-qPCR assays in a stepwise process to optimize the detection of COVID-19. Strategies: From August 2020 to November 2020, 43,399 sufferers had been screened in our laboratory for COVID-19 diagnostic by RT-qPCR utilizing nasopharyngeal swab. General, 4,691 of the 43,399 had been discovered to…
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Establishment of a duplex real-time qPCR method for detection of Salmonella spp. and Serratia fonticola in fishmeal
Salmonella spp. is a high-risk bacterial pathogen that’s monitored in imported animal-derived feedstuffs. Serratia fonticola is the bacterial species most often confused with Salmonella spp. in conventional identification strategies primarily based on biochemical traits, that are time-consuming and labor-intensive, and thus unsuitable for each day inspection and quarantine work. On this examine, we established a…
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DNA aptamers against bacterial cells can be efficiently selected by a SELEX process using state-of-the art qPCR and ultra-deep sequencing
DNA aptamers generated by cell-SELEX towards bacterial cells have gained elevated curiosity as novel and cost-effective affinity reagents for cell labelling, imaging and biosensing. Right here we describe the choice and identification of DNA aptamers for bacterial cells utilizing a mixed method based mostly on cell-SELEX, state-of-the-art functions of quantitative real-time PCR (qPCR), next-generation sequencing…
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