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Journal of Virological Methods

Jianing Chen, Chengyu Zhang, Yue Liu, Guangliang Liu
Disinfectant is commonly employed to eliminate infectious agents and prevent its transmission. In this study, we investigated the efficacy of Medilox(®) super-oxidized water on inactivating veterinary viruses mainly circulating in swine farms. The results demonstrated that this super-oxidized water could effectively inactivate porcine viruses.
January 10, 2017: Journal of Virological Methods
Spencer H Marshall, Raphael O Adegbola, Scott Adkins, Rayapati A Naidu
Tospoviruses (genus Tospovirus, family Bunyaviridae) are responsible for major losses in an extensive range of crops worldwide. New species of these single-stranded, ambisense RNA viruses regularly emerge and have been shown to maintain heterogeneous populations with individual isolates having quite variable biological and virulence characteristics. Most tospovirus phylogenetic studies have focused on analysis of a single gene, most often the nucleocapsid protein gene. Complete genomic RNA segment amplification as a single fragment would facilitate more detailed analyses of genome-wide sequence variability, but obtaining such sequences for a large number of tospovirus isolates using traditional methods of amplification and cloning of small overlapping fragments is tedious, time consuming and expensive...
January 9, 2017: Journal of Virological Methods
Satoshi Taniguchi, Aiko Fukuma, Hideki Tani, Shuetsu Fukushi, Masayuki Saijo, Masayuki Shimojima
Severe fever with thrombocytopenia syndrome (SFTS) is a recently-discovered, potentially fatal infectious disease caused by SFTS virus (SFTSV). Due to the inability of SFTSV to make clear cytopathic effects (CPE) in cell culture, titration and neutralization assays of the virus require immunostaining of inoculated cells; consequently, the assays are time-consuming and expensive. In this report, we demonstrate the use of a highly-passaged SFTSV strain, p50-2, in a neutralization assay, which made clear plaques in inoculated Vero cells under neutral red staining...
January 9, 2017: Journal of Virological Methods
Chui-Wan Lam, Sazaly AbuBakar, Li-Yen Chang
Nipah virus (NiV) is a highly pathogenic zoonotic paramyxovirus with unusual broad host tropism and is designated as a Category C pathogen by the U.S. National Institute of Allergy and Infectious Diseases. NiV infection is initiated after binding of the viral G glycoprotein to the host cell receptor. The aim of this study was to map the NiV G glycoprotein cell binding domain using a phage display system. The NiV G extracellular domain was truncated and displayed as attachment proteins on M13 phage g3p minor coat protein...
January 9, 2017: Journal of Virological Methods
Joany Castéra-Guy, Pierre-Alain Rubbo, Dramane Kania, Maud Lemoine, Philippe Van de Perre, Edouard Tuaillon
Antiviral therapy can be avoided during the low replicative phase of chronic Hepatitis B virus (HBV) infection which is characterized notably by HBV DNA concentration below 2000 IU/ml. Simplified diagnostic tests can improve access to HBV DNA monitoring in resource-limited settings. The capacity of a new semi-quantitative real-time PCR approach based on sample-to-standard relative detection of the target to discriminate samples with HBV DNA levels above or below the clinical threshold of 2000 IU/ml was compared to a quantitative assay (Roche CobasAmpliPrep/CobasTaqMan HBV Test v2...
January 6, 2017: Journal of Virological Methods
Selvaraj Tamilselvan, Thirunavukkarasu Ashokkumar, Kasivelu Govindaraju
In the present investigation, silver nanoparticles (AgNPs) interactions with Bombyx mori Nuclear Polyhedrosis virus (BmNPV) were characterized using High-Resolution Scanning Electron Microscopy (HR-SEM), Energy Dispersive X-ray Analysis (EDAX), Transmission Electron Microscopy (TEM), Atomic Force Microcopy (AFM) and Confocal Microscope (CM). HR-SEM study reveals that the biosynthesized AgNPs have interacted with BmNPV and were found on the surface. TEM micrographs of normal and viral polyhedra treated with AgNPs showed that the nanoparticles were accumulated in the membrane and it was noted that some of the AgNPs successfully penetrated the membrane by reaching the capsid of BmNPV...
January 5, 2017: Journal of Virological Methods
Shaun W Lim, Shea T Lance, Kenneth M Stedman, Adam R Abate
Characterizing virus-host relationships is critical for understanding the impact of a virus on an ecosystem, but is challenging with existing techniques, particularly for uncultivable species. We present a general, cultivation-free approach for identifying phage-associated bacterial cells. Using PCR-activated cell sorting, we interrogate millions of individual bacteria for the presence of specific phage nucleic acids. If the nucleic acids are present, the bacteria are recovered via sorting and their genomes analyzed...
December 29, 2016: Journal of Virological Methods
Tran Quang Huy, Nguyen Thi Hien Thanh, Nguyen Thanh Thuy, Pham Van Chung, Pham Ngoc Hung, Anh-Tuan Le, Nguyen Thi Hong Hanh
Silver nanoparticles (AgNPs) have been proven to have noticeable cytotoxicity in vitro and antiviral activity against some types of enveloped viruses. This paper presents the cytotoxicity and antiviral activity of pure AgNPs synthesized by the electrochemical method, towards cell culture and poliovirus (a non-enveloped virus). Prepared AgNPs were characterized by ultraviolet-visible spectroscopy, energy-dispersive X-ray spectroscopy and transmission electron microscopy. Before incubation with poliovirus, different concentrations of AgNPs were added to human rhabdomyosarcoma (RD) cell monolayers seeded in 96 well plates for testing their cytotoxicity...
December 28, 2016: Journal of Virological Methods
Susan Bennett, Rory N Gunson
Viral gastroenteritis is a major health problem with significant morbidity and economic consequences. Viral gastroenteritis is caused by a number of viruses, including norovirus, rotavirus, adenovirus, astrovirus, and sapovirus. Conventional diagnosis is based on direct antigen detection and electron microscopy, however enzyme immunoassay's are insensitive and not available for all relevant pathogens, and electron microscope (EM) is no longer routinely carried out in most laboratories. Most laboratories now offer norovirus real-time PCR testing however the availability of other assays is variable...
December 28, 2016: Journal of Virological Methods
Philipp Kaiser, Sunil K Joshi, Peggy Kim, Peilin Li, Hongbing Liu, Andrew P Rice, Joseph K Wong, Steven A Yukl
Despite intensive study, it is unclear which mechanisms are responsible for latent HIV infection in vivo. One potential mechanism is inhibition of HIV transcriptional elongation, which results in short abortive transcripts containing the trans-activation response (TAR) region. Because the relative levels of total (including short) and processive transcripts provide measures of HIV transcriptional initiation and elongation, there is a compelling need for techniques that accurately measure both. Nonetheless, prior assays for total transcripts have been semi-quantitative and have seen limited application to patient samples...
December 27, 2016: Journal of Virological Methods
Krister Melén, Laura Kakkola, Felix He, Kari Airenne, Olli Vapalahti, Helen Karlberg, Ali Mirazimi, Ilkka Julkunen
There is an urgent need for Ebola virus (EBOV) proteins, EBOV-specific antibodies and recombinant antigens to be used in diagnostics and as potential vaccine candidates. Our objective was to produce and purify recombinant proteins for immunological assays and for the production of polyclonal EBOV specific antibodies. In addition, a limited comparison of the adjuvant effects of Freund's complete adjuvant (FCA) and adjuvant system 03 (AS03) was carried out. Recombinant EBOV GST-VP24, -VP30, -VP35, -VP40 and -NP were produced in E...
December 23, 2016: Journal of Virological Methods
Matthew Frankel, Kenn Forberg, Kelly E Coller, Michael G Berg, John Hackett, Gavin Cloherty, George J Dawson
Human Pegivirus 2 (HPgV-2) was recently identified in the bloodstream of HCV-infected and multiply transfused individuals. Initial reports show HPgV-2 circulates at a low prevalence in HCV co-infected individuals, necessitating testing of large cohorts of samples to identify infected persons. The identification of additional HPgV-2 cases was facilitated by the development of a high throughput and reliable molecular reverse transcription polymerase chain reaction (RT-PCR) assay intended for use on the automated Abbott m2000 system with a capability of extracting and testing 96 samples at once...
December 22, 2016: Journal of Virological Methods
Brian J Kearney, Matthew A Voorhees, Priscilla L Williams, Scott P Olschner, Cynthia A Rossi, Randal J Schoepp
Viral preparations are essential components in diagnostic research and development. The production of large quantities of virus traditionally is done by infecting numerous tissue culture flasks or roller bottles, which require large incubators and/or roller bottle racks. The Corning HYPERFlask(®) is a multilayer flask that uses a gas permeable film to provide gas exchange between the cells and culture medium and the atmospheric environment. This study evaluated the suitability of the HYPERFlask for production of Lassa, Ebola, Bundibugyo, Reston, and Marburg viruses and compared it to more traditional methods using tissue culture flasks and roller bottles...
December 21, 2016: Journal of Virological Methods
Lyns Etienne, Poorval Joshi, Laura Dingle, Eugene Huang, Peter Grzesik, Prashant J Desai
Our laboratory was one of the first to engineer a live fluorescent tag, enhanced green fluorescent protein (eGFP), that marked the capsid of herpes simplex virus type 1 (HSV-1) and subsequently maturing virus as the particle made its way to the cell surface. In the present study we sought to increase the repertoire of colors available as fusion to the small capsid protein, VP26, so that they can be used alone or in conjunction with other fluorescent tags (fused to other HSV proteins) to follow the virus as it enters and replicates within the cell...
December 21, 2016: Journal of Virological Methods
A E Greijer, O Ramayanti, S A W M Verkuijlen, Z Novalić, H Juwana, J M Middeldorp
Epstein-Barr virus (EBV) is etiologically linked to multiple acute, chronic and malignant diseases. Detection of EBV-RNA transcripts in tissues or biofluids besides EBV-DNA can help in diagnosing EBV related syndromes. Sensitive EBV transcription profiling yields new insights on its pathogenic role and may be useful for monitoring virus targeted therapy. Here we describe a multi-gene quantitative RT-PCR profiling method that simultaneously detects a broad spectrum (n=16) of crucial latent and lytic EBV transcripts...
December 16, 2016: Journal of Virological Methods
Udeni B R Balasuriya, Pei-Yu Alison Lee, Yun-Long Tsai, Chuan-Fu Tsai, Yu-Han Shen, Hsiao-Fen Grace Chang, Ashley Skillman, Hwa-Tang Thomas Wang, Stéphane Pronost, Yan Zhang
Equine herpesvirus myeloencephalopathy (EHM), a major problem for the equine industry in the United States, is caused by equine herpesvirus-1 (EHV-1). In addition, EHV-1 is associated with upper respiratory disease, abortion, and chorioretinal lesions in horses. Here we describe the development and evaluation of an inexpensive, user-friendly insulated isothermal PCR (iiPCR) method targeting open reading 30 (ORF30) to detect both neuropathogenic and non-neuropathogenic strains on the field-deployable POCKIT™ device for point-of-need detection of EHV-1...
December 16, 2016: Journal of Virological Methods
Carole L Wallis, Raquel V Viana, Shanmugam Saravanan, Carlos Silva de Jesus, Clement Zeh, Elias K Halvas, John W Mellors
BACKGROUND: HIV-1 sequence variation is a major obstacle to developing molecular based assays for multiple subtypes. This study sought to independently assess performance characteristics of the ViroSeq™ HIV-1 Integrase RUO Genotyping Kit (Celera, US) for samples of multiple different HIV-1 subtypes. METHODS: 264 samples were tested in the validation, 106 from integrase inhibitor naïve patients' sent for routine HIV-1 drug resistance testing after failing a 1st- or 2nd-line regimen, and 158 samples from an external virology quality assurance program (VQA)...
December 16, 2016: Journal of Virological Methods
Rutger M Schepp, Guy A M Berbers, José A Ferreira, Johan H Reimerink, Fiona R van der Klis
Large-scale serosurveillance or vaccine studies for poliovirus using the "gold standard" WHO neutralisation test (NT) are very laborious and time consuming. With the polio eradication at hand and with the removal of live attenuated Sabin strains from the oral poliovirus vaccine (OPV), starting with type 2 (as of April 2016), laboratories will need to conform to much more stringent laboratory biosafety regulations when handling live poliovirus strains. In this study, a poliovirus binding inhibition multiplex immunoassay (polio MIA) using inactivated poliovirus vaccine (IPV-Salk) was developed for simultaneous quantification of serum antibodies directed to all three poliovirus types...
December 14, 2016: Journal of Virological Methods
L Elvira-González, A V Puchades, C Carpino, A Alfaro-Fernandez, M I Font-San-Ambrosio, L Rubio, L Galipienso
Southern tomato virus (STV) is a double stranded RNA (dsRNA) virus belonging to genus Amalgavirus (family Amalgamaviridae) which has been detected in tomato plants showing stunting, fruit discoloration and size reduction. A one-step reverse-transcription loop-mediated isothermal amplification (RT-LAMP) assay was developed for the detection of STV in total RNA or sap extracts (obtained just by grinding in buffer) from STV-infected tomato plants by using a set of three primers pairs which were designed to the sequence of the STV putative coat protein...
December 10, 2016: Journal of Virological Methods
Jaclyn A Miranda, Grieg F Steward
Reverse transcription, quantitative PCR (RT-qPCR) is a sensitive method for quantification of specific RNA targets, but the first step of the assay, reverse transcription, is notoriously variable and sensitive to reaction conditions. In this study, we used purified Bacteriophage MS2 genomic RNA as a model virus target to test two different RT enzymes (SuperScript II and SuperScript III), two RT-priming strategies (gene-specific primers and random hexamers), and varying background RNA concentrations (0-50ngμl(-1)) to determine how these variables influence the efficiency of reverse transcription over a range of target concentrations (10(1)-10(7) copies μl(-1))...
December 7, 2016: Journal of Virological Methods
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