Protocols in Current Issue
    Screening Method for CRISPR/Cas9 Inhibition of a Human DNA Virus: Herpes Simplex Virus
    [Abstract] The efficiency of cleavage of individual CRISPR/Cas9-sgRNAs remains difficult to predict based on the CRISPR target sequence alone. Different intracellular environments (dependent on cell type or cell cycle state for example) may affect sgRNA efficiency by altering accessibility of genomic DNA through DNA modifications such as epigenetic marks and ...
    Paper Lateral Flow Biosensor for Nodavirus Reverse Transcribed RNA Detection
    Authors:  Dimitra K. Toubanaki and Evdokia Karagouni, date: 08/05/2020, view: 385, Q&A: 0
    [Abstract] Paper nanobiosensors have been established as an excellent platform for analysis of veterinary and human pathogens causing various diseases. Especially, lateral flow assays or biosensors ideal for sensitive, rapid, robust and accurate analysis in laboratory setups and on-site analysis. Viral RNA detection is of great importance for public health ...
    Safe DNA-extraction Protocol Suitable for Studying Tree-fungus Interactions
    Authors:  Susanna Keriö, Eeva Terhonen and Jared M. LeBoldus, date: 06/05/2020, view: 1431, Q&A: 0
    [Abstract] We present a safe and low-cost method suitable for DNA extraction from mycelium and tree tissue samples. After sample preparation, the extraction takes about 60 min. Method performance was tested by extracting DNA from various tree tissue samples and from mycelium grown on solid and liquid media. DNA was extracted from juvenile and mature host ...
    Viral Double-Stranded RNA Detection by DNase I and Nuclease S1 digestions in Leishmania parasites
    Authors:  Nathalie Isorce and Nicolas Fasel, date: 05/05/2020, view: 703, Q&A: 0
    [Abstract] Many RNA viruses are found in protozoan parasites. They can be responsible for more serious pathology or treatment failure. For the detection of viral double-stranded RNA (dsRNA), sequence-dependent and -independent methods are available, such as quantitative real-time PCR and immunofluorescence, dot blot, ELISA or sequencing. The technique ...
    Localizing Genome Segments and Protein Products of a Multipartite Virus in Host Plant Cells
    Authors:  Marie-Stephanie Vernerey, Elodie Pirolles, Stephane Blanc and Anne Sicard, date: 12/05/2019, view: 1441, Q&A: 0
    [Abstract] A founding paradigm in virology is that the spatial unit of the viral replication cycle is an individual cell. This concept applied to multipartite viruses–which have a genome composed of two or more nucleic acid segments, each individually encapsulated–implies that all segments constituting a viral genome need to coinfect the same host cell for ...
    Leishmania Parasite Quantification by Bioluminescence in Murine Models
    Authors:  Marta Reverte and Nicolas Fasel, date: 11/20/2019, view: 1454, Q&A: 0
    [Abstract] Leishmaniasis remains a major public health problem worldwide with a prevalence of 12 million, an incidence of 1 million persons, and 350 million people being at risk. Murine models have been largely used for studying the host-pathogen relationship and developing effective chemotherapies against Leishmania parasites. Thus, preclinical ...
    A Mismatch-tolerant RT-LAMP Method for Molecular Diagnosis of Highly Variable Viruses
    Authors:  Yingxue Li, Yi Zhou, Yingying Ma, Rong Xu, Xia Jin and Chiyu Zhang, date: 11/05/2019, view: 2089, Q&A: 0
    [Abstract] Loop-mediated isothermal amplification (LAMP) has been widely used in the detection of pathogens. However, there are usually numerous variants in one viral pathogen and primers employed in LAMP can hardly match all these variants. The mismatches between the primers and the viral genomes, especially those at the 3′-end of the primers, hinder LAMP ...
    Solid Phase PCR on 3D Microstructure ArrayChip for Pathogen Detection Application
    Authors:  Krishna Kant and Tien Anh Ngo, date: 08/05/2019, view: 1819, Q&A: 0
    [Abstract] Advanced free angle photolithography (FAPL) is presented for making 3D supercritical angle fluorescence (SAF) microstructures and transfer them on to polymeric chips using injection molding technique for low-cost microfluidic devices embedded with optical sensing structures. A solid phase polymerase chain reaction (SP-PCR) is used as model ...

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