Electron microscopy: essentials for viral structure, morphogenesis and rapid diagnosis

Zhang, Ying; Hung, Tao; Song, Jingdong; He, Jin‐Sheng

doi:10.1007/s11427-013-4476-2

Cited by 31 publications

(24 citation statements)

References 86 publications

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“…Despite the fact that TEM analysis is a very useful means by which to visualize virus particles, this technique comes with certain limitations such as lengthy sample preparation procedures, expensive equipment and specialized expertise (Goldsmith and Miller, 2009; Vale et al, 2010; Zhang et al, 2013; Brown et al, 2015). Thus, only a relatively small number of strains were subjected to TEM analysis.…”

Section: Discussionmentioning

confidence: 99%

Detecting Lactococcus lactis Prophages by Mitomycin C-Mediated Induction Coupled to Flow Cytometry Analysis

et al. 2017

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Most analyzed Lactococcus lactis strains are predicted to harbor one or more prophage genomes within their chromosome; however, the true extent of the inducibility and functionality of such prophages cannot easily be deduced from sequence analysis alone. Chemical treatment of lysogenic strains with Mitomycin C is known to cause induction of temperate phages, though it is not always easy to clearly identify a lysogenic strain or to measure the number of released phage particles. Here, we report the application of flow cytometry as a reliable tool for the detection and enumeration of released lactococcal prophages using the green dye SYTO-9.

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Section: Discussionmentioning

confidence: 99%

Detecting Lactococcus lactis Prophages by Mitomycin C-Mediated Induction Coupled to Flow Cytometry Analysis

et al. 2017

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“…Furthermore, even with the high resolution afforded by TEM, it can be difficult to distinguish true viruses from non-virus particles of similar size, as will typically be found in environmental samples (i.e., non-viral or 'fake' VLPs). For an in-depth overview of TEM capabilities for viruses, see [168].…”

Section: Efficiency Of Virus Resuspension From Soilsmentioning

confidence: 99%

Coming-of-Age Characterization of Soil Viruses: A User’s Guide to Virus Isolation, Detection within Metagenomes, and Viromics

et al. 2020

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The study of soil viruses, though not new, has languished relative to the study of marine viruses. This is particularly due to challenges associated with separating virions from harboring soils. Generally, three approaches to analyzing soil viruses have been employed: (1) Isolation, to characterize virus genotypes and phenotypes, the primary method used prior to the start of the 21st century. (2) Metagenomics, which has revealed a vast diversity of viruses while also allowing insights into viral community ecology, although with limitations due to DNA from cellular organisms obscuring viral DNA. (3) Viromics (targeted metagenomics of virus-like-particles), which has provided a more focused development of ‘virus-sequence-to-ecology’ pipelines, a result of separation of presumptive virions from cellular organisms prior to DNA extraction. This separation permits greater sequencing emphasis on virus DNA and thereby more targeted molecular and ecological characterization of viruses. Employing viromics to characterize soil systems presents new challenges, however. Ones that only recently are being addressed. Here we provide a guide to implementing these three approaches to studying environmental viruses, highlighting benefits, difficulties, and potential contamination, all toward fostering greater focus on viruses in the study of soil ecology.

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“…Clinical microbiologists are constantly striving to deliver accurate test results more quickly to ensure that appropriate treatment and infection control measures are implemented on time. Conventional diagnostic methods, such as microscopy and culture are laborious, timeconsuming and require considerable technical skill for accurate analysis of the pathogen [1,2]. The technology revolution that has occurred over the last few decades has considerably improved and expanded the diagnostic capabilities of modern clinical microbiology laboratories.…”

Section: Introductionmentioning

confidence: 99%

Amplification chemistries in clinical virology

Dunbar

Das

2019

Journal of Clinical Virology

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Molecular diagnostic methods have evolved and matured considerably over the last several decades and are constantly being evaluated and adopted by clinical laboratories for the identification of infectious pathogens. Advancement in other technologies such as fluorescence, electronics, instrumentation, automation, and sensors have made the overall diagnostic process more accurate, sensitive, and rapid. Nucleic acid based detection procedures, which rely on the fundamental principles of DNA replication have emerged as a popular and standard diagnostic method, and several commercial assays are currently available based on different nucleic acid amplification techniques. This review focuses on the major amplification chemistries that are used for developing commercial assays and discusses their application in the clinical virology laboratory. Polymerase chain reaction (PCR)The development of PCR by Kary Mullis in the 1980s revolutionized the detection of microbial pathogens in clinical specimens. PCR is a highly sensitive method that involves cyclical heating and cooling of the reaction mixture to facilitate amplification of the target DNA [3]. The method consists of three basic steps (denaturation, annealing, and extension) and the reaction mixture requires four primary components (template DNA, primers, nucleotides, and a DNA polymerase) (Fig. 1a). Double-stranded target DNA (dsDNA) is thermally denatured to form a single-stranded DNA template (ssDNA). Oligonucleotide primers anneal to complementary target sequences on the nucleic acid template and are extended by the DNA polymerase, thus creating the resultant PCR product (amplicon). The reaction occurs in a programmable thermal https://doi.

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Electron microscopy: essentials for viral structure, morphogenesis and rapid diagnosis

Cited by 31 publications

References 86 publications

Detecting Lactococcus lactis Prophages by Mitomycin C-Mediated Induction Coupled to Flow Cytometry Analysis

Detecting Lactococcus lactis Prophages by Mitomycin C-Mediated Induction Coupled to Flow Cytometry Analysis

Coming-of-Age Characterization of Soil Viruses: A User’s Guide to Virus Isolation, Detection within Metagenomes, and Viromics

Amplification chemistries in clinical virology

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