International Journal of Molecular Sciences

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Dr. Ilka Engelmann

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Laboratoire de Virologie, CHU de Montpellier, Faculté de Médecine, Université de Montpellier, Montpellier, France
Interests: molecular diagnostics of viruses; markers of viral infection; viral infections in immunocompromised host; enterovirus; microRNAs in viral infections; viral persistance
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Dear Colleagues,

Molecular biology has revolutionized virology in several aspects. Due to the limitations of viral culture, antigen detection, and serology, the molecular detection of viral nucleic acids has become the method of choice for the diagnosis of viral infections in many circumstances. Molecular epidemiology of viruses has led to a better understanding of their phylogenetic relationships, which has led to changes in the taxonomy of several virus families. The recent development of rapid molecular techniques enhances the impact on viral diagnostics for patient management. Next generation sequencing is now entering clinical laboratories to permit the diagnosis of a previously unachieved large spectrum of viruses.

This Special Issue on "Molecular detection and typing of viruses" will include manuscripts on recent advancements in molecular biological techniques in the field of virology. Papers aiming to improve our understanding of epidemiology, diagnosis, and management of viral infections are welcome. Original research and reviews on these and related topics are invited.

You may choose our Joint Special Issue in Diagnostics.

Dr. Ilka Engelmann
Guest Editor

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Keywords

PCR
molecular biology
typing
epidemiology
sequencing
molecular diagnostics

Published Papers (2 papers)

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Research

13 pages, 985 KiB

Open AccessArticle

Swift and Reliable “Easy Lab” Methods for the Sensitive Molecular Detection of African Swine Fever Virus

by Ahmed Elnagar, Jutta Pikalo, Martin Beer, Sandra Blome and Bernd Hoffmann

Int. J. Mol. Sci. 2021, 22(5), 2307; https://doi.org/10.3390/ijms22052307 - 25 Feb 2021

Cited by 10 | Viewed by 3985

Abstract

African swine fever (ASF) is a contagious viral hemorrhagic disease of domestic pigs and wild boars. The disease is notifiable to the World Organisation for Animal Health (OIE) and is responsible for high mortality and serious economic losses. PCR and real-time PCR (qPCR) are the OIE-recommended standard methods for the direct detection of African swine fever virus (ASFV) DNA. The aim of our work was the simplification and standardization of the molecular diagnostic workflow in the lab. For validation of this “easy lab” workflow, different sample materials from animal trials were collected and analyzed (EDTA blood, serum, oral swabs, chewing ropes, and tissue samples) to identify the optimal sample material for diagnostics in live animals. Based on our data, the EDTA blood samples or bloody tissue samples represent the best specimens for ASFV detection in the early and late phases of infection. The application of prefilled ready-to-use reagents for nucleic acid extraction or the use of a Tissue Lysis Reagent (TLR) delivers simple and reliable alternatives for the release of the ASFV nucleic acids. For the qPCR detection of ASFV, different published and commercial kits were compared. Here, a lyophilized commercial kit shows the best results mainly based on the increased template input. The good results of the “easy lab” strategy could be confirmed by the ASFV detection in field samples from wild boars collected from the 2020 ASFV outbreak in Germany. Appropriate internal control systems for extraction and PCR are key features of the “easy lab” concept and reduce the risk of false-negative and false-positive results. In addition, the use of easy-to-handle machines and software reduces training efforts and the misinterpretation of results. The PCR diagnostics based on the “easy lab” strategy can realize a high sensitivity and specificity comparable to the standard PCR methods and should be especially usable for labs with limited experiences and resources. Full article

(This article belongs to the Special Issue Molecular Detection and Typing of Viruses)

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11 pages, 1491 KiB

Open AccessArticle

Optimized qRT-PCR Approach for the Detection of Intra- and Extra-Cellular SARS-CoV-2 RNAs

by Tuna Toptan, Sebastian Hoehl, Sandra Westhaus, Denisa Bojkova, Annemarie Berger, Björn Rotter, Klaus Hoffmeier, Jindrich Cinatl, Jr., Sandra Ciesek and Marek Widera

Int. J. Mol. Sci. 2020, 21(12), 4396; https://doi.org/10.3390/ijms21124396 - 20 Jun 2020

Cited by 57 | Viewed by 7546

Abstract

The novel coronavirus SARS-CoV-2 is the causative agent of the acute respiratory disease COVID-19, which has become a global concern due to its rapid spread. Meanwhile, increased demand for testing has led to a shortage of reagents and supplies and compromised the performance of diagnostic laboratories in many countries. Both the World Health Organization (WHO) and the Center for Disease Control and Prevention (CDC) recommend multi-step RT-PCR assays using multiple primer and probe pairs, which might complicate the interpretation of the test results, especially for borderline cases. In this study, we describe an alternative RT-PCR approach for the detection of SARS-CoV-2 RNA that can be used for the probe-based detection of clinical isolates in diagnostics as well as in research labs using a low-cost SYBR green method. For the evaluation, we used samples from patients with confirmed SARS-CoV-2 infections and performed RT-PCR assays along with successive dilutions of RNA standards to determine the limit of detection. We identified an M-gene binding primer and probe pair highly suitable for the quantitative detection of SARS-CoV-2 RNA for diagnostic and research purposes. Full article

(This article belongs to the Special Issue Molecular Detection and Typing of Viruses)

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