An assessment of the reproducibility of reverse transcription digital PCR quantification of HIV-1

SARS-CoV-2 Subgenomic RNAs: Characterization, Utility, and Perspectives

“…Limited yields of cells or fluids from sampling procedures such as nasopharyngeal swabs, and the presence of potential inhibitors (e.g., chemical or protein contaminants) in clinical samples, require that PCR amplification and detection be highly sensitive and reliable during SARS-CoV-2 nucleic acid analysis. Digital PCR has demonstrated significant advantages in both SARS-CoV-2 gRNA [94][95][96] and sgRNA [49,97,98] studies due to its ability for absolute quantification [98][99][100][101][102][103][104][105], tolerance to inhibitors [106], increased precision at low analyte copy numbers [107][108][109], and inter-run reproducibility [110][111][112]. One additional distinct advantage of the digital PCR approach is its lower susceptibility to sequence mismatches, which is especially relevant as emerging mutations that can potentially predominate could affect the performance of real-time PCR-based assays if they occur in regions where the PCR primer and probes are located [113,114].…”

An ultra‐sensitive and highly specific detection method for a newly discovered Oncotshavirus using one‐step reverse transcription droplet‐based digital PCR

“…This species is an economically important fish variety, and our detection method will provide an important assay for food safety of aquatic products. Although the ddPCR has been widely used in both DNA and RNA virus detection (Amoroso et al, 2021;Falak et al, 2021;Gourinat et al, 2015;Jiang et al, 2020;Pavšič et al, 2016;Pinheiro-de-Oliveira et al, 2019;Tan et al, 2021), there is no record that the method has been used on aquatic virus detection previously. The one-step ddPCR detection method established in this study is the first attempt to apply the assay to aquatic virus, which is meaningful to the detection methodology of aquatic virus.…”

“…Given the advantage of better sensitivity and amplification efficiency than traditional qPCR methods, dPCR develops rapidly and has been applied to absolute quantitation (Garcia-Murillas et al, 2013), copy number determination (Hindson et al, 2011) and single molecule identification (Fu et al, 2015;Zhu et al, 2012). Although dPCR has achieved wide application among the quantitative detection of viruses (Amoroso et al, 2021;Falak et al, 2021;Jiang et al, 2020;Pavšič et al, 2016;Pinheiro-de-Oliveira et al, 2019;Tan et al, 2021), it has not been applied to the detection of aquatic virus in previous study. Moreover, the reverse transcription steps necessary to evaluate an RNA virus could cause significant deviations to the final quantitative evaluation results.…”

Metrological framework to support accurate, reliable, and reproducible nucleic acid measurements

“…Similarly in the frame of Infect-Met and AntiMicroResist projects, methods for reliable quantification of several infectious agents, human influenza A virus, human cytomegalovirus, Mycobacterium tuberculosis, and HIV, and their resistance were investigated [90][91][92][93][94][95][96][97]. One of the measurement procedures developed and evaluated in these projects, namely quantification of DNA extracted from human cytomegalovirus, has been also listed in the JCTLM database.…”