Effective RNA extraction with readily available reagents: A comparative analysis of alternative protocols.

J Virol Methods

Respiratory Viruses, Exanthemateous, Enterovirus and Viral Emergencies Laboratory, Oswaldo Cruz Institute/Fiocruz, Av. Brasil 4365, Rio de Janeiro, RJ 21040-360, Brazil.

Published: December 2025


Category Ranking

98%

Total Visits

921

Avg Visit Duration

2 minutes

Citations

20

Article Abstract

The SARS-CoV-2 pandemic exposed significant challenges in diagnostic laboratory workflows, where shortages of reagents and automation worsened the management of surges in testing demand. This study evaluates four direct sample preparation protocols and proposes an economical alternative RNA extraction method using generic silica columns as a feasible solution during resource scarcity. While the non-extraction protocols tested presented an elevated risk of producing false negatives, the alternative method demonstrated results statistically comparable to standard manual and automated extraction protocols (ANOVA, F=0.69, p = 0.415), with an average Ct difference of only one cycle. The protocol demonstrated reduced efficiency only at extreme viral loads and offers a cost-effective, accessible option for public health laboratories facing supply chain constraints during increasingly common epidemic pressures.

Download full-text PDF

Source
http://dx.doi.org/10.1016/j.jviromet.2025.115242DOI Listing

Publication Analysis

Top Keywords

rna extraction
8
effective rna
4
extraction reagents
4
reagents comparative
4
comparative analysis
4
analysis alternative
4
protocols
4
alternative protocols
4
protocols sars-cov-2
4
sars-cov-2 pandemic
4

Similar Publications

Background: The dysregulation of long-chain noncoding RNAs (lncRNAs) causes several complex human diseases including neurodegenerative disorders across the globe.

Methods And Results: This study aimed to investigate lncRNA expression profiles of Withania somnifera (WS)-treated human neuroblastoma SK-N-SH cells at different timepoints (3 & 9 h) and concentrations (50 & 100 µg/mL) using RNA sequencing. Differential gene expression analysis showed a total of 4772 differentially expressed lncRNAs, out of which 3971 were upregulated and 801 were downregulated compared to controls.

View Article and Find Full Text PDF

Unlabelled: There is a need for the development of broad-spectrum antiviral compounds that can act as first-line therapeutic countermeasures to emerging viral infections. Host-directed approaches present a promising avenue of development and carry the benefit of mitigating risks of viral escape mutants. We have previously found the SKI (super killer) complex to be a broad-spectrum, host-target with our lead compound ("UMB18") showing activity against influenza A virus, coronaviruses, and filoviruses.

View Article and Find Full Text PDF

Crab shell polypeptides enhance calcium dynamics and osteogenic activity in osteoporosis.

Front Pharmacol

August 2025

Department of Genetics and Cell Biology, School of Basic Medicine, Qingdao University, Qingdao, Shandong, China.

Background: Osteoporosis (OP) is a chronic, systemic skeletal disorder characterized by progressive bone loss and microarchitectural deterioration, which increases fracture susceptibility and presents a challenging set of global healthcare problems. Current pharmacological interventions are limited by adverse effects, high costs, and insufficient long-term efficacy. Here, we identify snow crab shell-derived polypeptides (SCSP) as a potent osteoprotective agent.

View Article and Find Full Text PDF

Background And Aim: is a significant zoonotic pathogen linked to reproductive losses in livestock and serious health risks in humans. In Iraq, listeriosis remains underreported in sheep, with limited data on its molecular and epidemiological characteristics. This study aimed to (1) estimate the seroprevalence of in recently aborted ewes, (2) evaluate the association between seropositivity and clinical indicators, and (3) perform molecular detection and phylogenetic analysis of polymerase chain reaction (PCR)-confirmed isolates.

View Article and Find Full Text PDF

Purpose: To verify the stability and reliability of circulating microRNA (miRNA) profiles in plasma and serum under different processing and storage conditions to inform future applications to circulating biomarker analyses.

Background: The development of blood-based methods for early disease detection has become increasingly desirable across various medical fields. RNA profiles have been investigated but have been a challenge due to rapid degradation of the analyte by ubiquitous RNases.

View Article and Find Full Text PDF