A rapid and simple quantitative method for specific Detection of Smaller Co-terminal RNA by PCR (DeSCo-PCR): Application to the detection of viral subgenomic RNAs.

A rapid and simple quantitative method for specific Detection of Smaller Co-terminal RNA by PCR (DeSCo-PCR): Application to the detection of viral subgenomic RNAs. RNA. 2020 Apr 01;: Authors: Kanodia P, Prasanth KR, Roa-Linares VC, Bradrick SS, Garcia-Blanco MA, Miller WA Abstract RNAs that are 5'-truncated versions of a longer RNA, but share the same 3' terminus can be generated by alternative promoters in transcription of cellular mRNAs or by replicating RNA viruses. These truncated RNAs cannot be distinguished from the longer RNA by a simple two-primer RT-PCR because primers that anneal to the cDNA from the smaller RNA also anneal to - and amplify - the longer RNA-derived cDNA. Thus, laborious methods, such as northern blot hybridization, are used to distinguish shorter from longer RNAs. For rapid, low-cost and specific detection of these truncated RNAs, we report Detection of Smaller Co-terminal RNA by PCR (DeSCo-PCR). DeSCo-PCR employs a non-extendable blocking primer (BP), which outcompetes a forward primer (FP) for annealing to longer RNA-derived cDNA, while FP outcompetes BP for annealing to shorter RNA-derived cDNA. In the presence of BP, FP and the reverse primer, only cDNA from the shorter RNA is amplified in a single-tube reaction containing both RNAs. Many positive strand RNA viruses generate 5'-truncated forms of the genomic RNA (gRNA) called subgenomic RNAs (sgRNA), which play key roles in viral gene expression and pathogenicity. We demons...
Source: RNA - Category: Genetics & Stem Cells Authors: Tags: RNA Source Type: research

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Abstract The emergence and re-emergence of Zika virus (ZIKV), is a cause for international concern. These highly pathogenic arboviruses represent a serious health burden in tropical and subtropical areas worldwide. Despite these burdens, antiviral therapies do not exist, and inhibitors of ZIKV are therefore urgently needed. To elucidate the anti-ZIKV effect of lycorine, we used reverse transcription-quantitative real-time PCR (qRT-PCR), immunofluorescence, Westernwestern blot, and plaque forming assay to analyse viral RNA (vRNA), viral protein, progeny virus counts, and validated inhibitors in vitro using a variet...
Source: Virology - Category: Virology Authors: Tags: Virology Source Type: research
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Source: Medgadget - Category: Medical Devices Authors: Tags: Exclusive Medicine Public Health Source Type: blogs
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Source: International Journal of Environmental Research and Public Health - Category: Environmental Health Authors: Tags: Review Source Type: research
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Source: IPS Inter Press Service - Health - Category: International Medicine & Public Health Authors: Tags: Aid Economy & Trade Featured Global Headlines Health Humanitarian Emergencies TerraViva United Nations Jomo Kwame Sundaram & Anis Chowdhury Source Type: news
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Source: Viruses - Category: Virology Authors: Tags: Article Source Type: research
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Source: Viruses - Category: Virology Authors: Tags: Article Source Type: research
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Source: EurekAlert! - Infectious and Emerging Diseases - Category: Infectious Diseases Source Type: news
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Source: Pediatric Neurology - Category: Neurology Authors: Tags: Review Article Source Type: research
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Source: Viruses - Category: Virology Authors: Tags: Article Source Type: research
AbstractThe Japanese encephalitis virus (JEV) is one of the vector borne causes of encephalitis found in southeastern Asia. This positive single-stranded RNA virus is a member of the Flaviviridae family, which notably includes dengue, tick-borne, West Nile, Zika as well as yellow fever, and transmits to humans by infected mosquitos. The main site of interactions for antibodies against this virus is the envelope protein domain III (ED3). The present report investigates the time-dependent structural and conformational changes of JEV ED3 functional epitopes and escape mutants by computer simulations. The results indicate the ...
Source: Immunologic Research - Category: Allergy & Immunology Source Type: research
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