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Journal of Virological Methods

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https://www.readbyqxmd.com/read/28315718/absolute-quantification-of-cassava-brown-streak-virus-mrna-by-real-time-qpcr
#1
Rudolph R Shirima, Daniel Maeda, Edward Kanju, Gloria Ceasar, Flora I Tibazarwa, James P Legg
Cassava brown streak disease (CBSD) is the most important virus disease of cassava and a major food security threat in Africa. Yearly economic losses of up to $100 million USD have been attributed to CBSD. The lack of information on plant-virus interactions has restricted progress in breeding for CBSD resistance. Virus quantification is becoming a major tool for the quick and reliable assessment of plant host resistance. Therefore, a protocol for specific absolute quantification of Cassava brown streak virus (CBSV) and Ugandan cassava brown streak virus (UCBSV) was developed...
March 15, 2017: Journal of Virological Methods
https://www.readbyqxmd.com/read/28315717/exploring-novel-sero-epidemiological-tools-effect-of-different-storage-conditions-on-longitudinal-stability-of-microarray-slides-comprising-influenza-a-measles-and-streptococcus-pneumoniae-antigens
#2
Gudrun Freidl, Erwin de Bruin, Maarten Schipper, Marion Koopmans
In this study we evaluated the long-term stability of a microarray-based serological screening platform, containing antigens of influenza A, measles and Streptococcus pneumoniae, as part of a preparedness research program aiming to develop assays for syndromic disease detection. Spotted microarray slides were kept at four different storage regimes with varying temperature and humidity conditions. We showed that under the standard storage condition in a temperature-controlled (21°C) and desiccated environment (0% relative humidity), microarray slides remained stable for at least 22 months without loss of antigen quality, whereas the other three conditions (37°C, desiccated; Room temperature, non-desiccated; Frozen, desiccated) produced acceptable results for some antigens (influenza A, S...
March 15, 2017: Journal of Virological Methods
https://www.readbyqxmd.com/read/28315379/magnetic-nanoparticles-for-efficient-cell-transduction-with-semliki-forest-virus
#3
Baiba Kurena, Aleksandra Vežāne, Dace Skrastiņa, Olga Trofimova, Anna Zajakina
Semliki Forest virus (SFV) is a potential cancer gene therapy vector capable of providing high and transient expression of heterologous proteins in mammalian cells. However, SFV has shown suboptimal transduction levels in several cancer cell types as well as wide biodistribution of SFV has been observed after in vivo applications. Magnetic nanoparticles (MNPs) have been shown to increase cell transduction with several viral vectors in vitro under an external magnetic field and enhance magnetically guided viral vector delivery...
March 14, 2017: Journal of Virological Methods
https://www.readbyqxmd.com/read/28300606/one-step-multiplex-rt-qpcr-detects-three-citrus-viroids-from-different-genera-in-a-wide-range-of-hosts
#4
Fatima Osman, Tyler Dang, Sohrab Bodaghi, Georgios Vidalakis
A one-step multiplex reverse transcription real-time quantitative polymerase chain reaction (RT-qPCR) based on species-specific minor groove binding (MGB) probes, was developed for the simultaneous detection, identification, and quantification of three citrus viroids belonging to different genera. Citrus exocortis viroid (Pospiviroid), Hop stunt viroid (Hostuviroid), and Citrus bark cracking viroid (Cocadviroid) cause a variety of maladies in agriculturally significant crops. Therefore, reliable assays for their detection are essential tools for various government and industry organizations implementing disease management programs...
March 11, 2017: Journal of Virological Methods
https://www.readbyqxmd.com/read/28257802/assessment-of-influenza-a-neuraminidase-subtype-n1-potency-by-elisa
#5
Hongquan Wan, Ishrat Sultana, Laura K Couzens, Samuel Mindaye, Maryna C Eichelberger
Antibodies that inhibit neuraminidase (NA) activity of influenza virus provide resistance against disease and have been associated with milder epidemics. Although studies have demonstrated a correlation between NA inhibition antibody titers and vaccine efficacy, neither the quantity nor form of NA is measured in seasonal and pandemic influenza vaccines. In this report, we describe development of enzyme-linked immunosorbent assays (ELISAs) that are suitable for quantitation of the native form of NA of subtype N1...
March 10, 2017: Journal of Virological Methods
https://www.readbyqxmd.com/read/28284976/recombinant-vp1-protein-as-a-potential-marker-for-the-diagnosis-of-acute-hepatitis-a-virus-infection
#6
Haroldo Cid da Silva, Edimilson Domingos da Silva, Lia Laura Lewis-Ximenez de Souza Rodrigues, Marco Alberto Medeiros
Since hepatitis A virus (HAV) production is time-consuming and expensive, the use of recombinant proteins may represent an alternative source of antigens for diagnostic purposes. The present study aimed to express, purify and evaluate the potential of recombinant VP1 protein (rVP1) as a marker for the diagnosis of acute HAV infection. The rVP1 was expressed and purified successfully from Escherichia coli. The purified rVP1 was used to establish an in-house enzyme-linked immunosorbent assay (ELISA-rVP1) for detection of IgM antibodies in sera from HAV-positive patients...
March 8, 2017: Journal of Virological Methods
https://www.readbyqxmd.com/read/28274745/a-real-time-rt-qpcr-assay-for-the-detection-of-citrus-tatter-leaf-virus
#7
S J Cowell, S J Harper, W O Dawson
Citrus tatter leaf virus (CLTV) is globally distributed wherever citrus is grown, and, given the extensive use of CTLV sensitive rootstock, has the potential to be a significant threat to the citrus industry. In order to facilitate fast and reliable detection of this virus, we have developed a CTLV-specific real-time RT-qPCR assay. The optimized assay was found to be more reliable and sensitive compared to ELISA and end-point RT-PCR, detecting CTLV in up to 70% more plants. The real-time RT-qPCR is also specific, as it did not cross-react with the closely related Apple stem grooving virus or with the host itself; robust, being able to detect CTLV in young and mature host tissue types; and rapid...
March 6, 2017: Journal of Virological Methods
https://www.readbyqxmd.com/read/28274744/performance-of-the-samba-i-and-ii-hiv-1-semi-q-tests-for-viral-load-monitoring-at-the-point-of-care
#8
Neha Goel, Allyson V Ritchie, Sekesai Mtapuri-Zinyowera, Clement Zeh, Tetiana Stepchenkova, Jesse Lehga, Annemiek De Ruiter, Laura E Farleigh, Daniel Edemaga, Rosario So, Hiroshi Sembongi, Craig Wisniewski, Lourdes Nadala, Marco Schito, Helen Lee
Although access to antiretroviral therapy for HIV infection is increasing in resource-poor countries, viral load testing for monitoring of treatment efficacy remains limited, expensive, and confined to centralized laboratories. The SAMBA HIV-1 Semi-Q Test is a nucleic acid-based amplification assay developed for viral load monitoring performed on either the semi-automated SAMBA I system for laboratory use or the fully automated SAMBA II system for point-of care use. We have assessed the performance characteristics of the SAMBA HIV-1 Semi-Q Test on SAMBA I and SAMBA II systems according to the Common Technical Specifications of the European Community's 98/79 In Vitro Diagnostic Medical Devices Directive...
March 6, 2017: Journal of Virological Methods
https://www.readbyqxmd.com/read/28263762/development-and-evaluation-of-taqman-real-time-pcr-assay-for-detection-of-beak-and-feather-disease-virus
#9
Lenka Černíková, Eliška Vitásková, Alexander Nagy
Psittacine beak and feather disease (PBFD) is one of the most significant viral diseases in psittacine birds. The aim of the presented study was to develop a highly specific and sensitive TaqMan real-time PCR assay for universal detection of beak and feather disease virus (BFDV). Primers and a hydrolysis probe were selected on the highly conserved regions belonging to the ORF1 of the BFDV genome which were identified by aligning 814 genomic sequences downloaded from the GenBank database. The evaluation of the reaction parameters suggested a reaction efficiency of 97...
March 2, 2017: Journal of Virological Methods
https://www.readbyqxmd.com/read/28257801/analysis-of-bzlf1-mrna-detection-in-saliva-as-a-marker-for-active-replication-of-epstein-barr-virus
#10
Ursula Fagin, Linda Nerbas, Bastian Vogl, Wolfram J Jabs
Monitoring replicative Epstein-Barr virus (EBV) infection still remains a challenge in modern laboratory routine. The immediate-early protein BZLF1 mediates the switch between latent and replicate forms of EBV infection. The aim of this study was to analyze the feasibility of BZLF1 mRNA detection in saliva as a marker for active replication of the virus. Various specimens (saliva, plasma, PBMC) from 17 patients with EBV-induced infectious mononucleosis (IM) and 4 control patients were examined for expression of viral BZLF1 mRNA by means of real-time PCR...
February 28, 2017: Journal of Virological Methods
https://www.readbyqxmd.com/read/28254680/enhancing-the-sensitivity-of-dengue-virus-serotype-detection-by-rt-pcr-among-infected-children-in-india
#11
Syed Fazil Ahamed, Rosario Vivek, Shalini Kotabagi, Kaustuv Nayak, Anmol Chandele, Murali-Krishna Kaja, Anita Shet
Dengue surveillance relies on reverse transcription-polymerase chain reaction (RT-PCR), for confirmation of dengue virus (DENV) serotypes. We compared efficacies of published and modified primer sets targeting envelope (Env) and capsid-premembrane (C-prM) genes for detection of circulating DENV serotypes in southern India. Acute samples from children with clinically-diagnosed dengue were used for RT-PCR testing. All samples were also subjected to dengue serology (NS1 antigen and anti-dengue-IgM/IgG rapid immunochromatographic assay)...
February 28, 2017: Journal of Virological Methods
https://www.readbyqxmd.com/read/28242293/lyophilized-visually-readable-loop-mediated-isothermal-reverse-transcriptase-nucleic-acid-amplification-test-for-detection-ebola-zaire-rna
#12
Christoph Carter, Kevan Akrami, Drew Hall, Davey Smith, Eliah Aronoff-Spencer
Recent viral outbreaks highlight the need for reliable, yet broadly deployable diagnostics for detection of epidemic and emerging pathogens. In this study we designed and optimized methods to visually detect viral nucleic acid by isothermal amplification and SYBR dye intercalation. We designed and tested loop-mediated isothermal amplification (LAMP) primers and lyophilized reactions to optimize the detection of Zaire Ebola Virus (ZEBOV) and further evolved the LAMP platform to allow room-temperature storage for deployment in resource limited settings...
February 24, 2017: Journal of Virological Methods
https://www.readbyqxmd.com/read/28219761/development-and-validation-of-a-real-time-reverse-transcription-pcr-assay-for-rapid-and-low-cost-genotyping-of-hepatitis-c-virus-genotypes-1a-1b-2-and-3a
#13
Andrea D Olmstead, Tracy D Lee, Ron Chow, Kingsley Gunadasa, Brian Auk, Mel Krajden, Agatha N Jassem
Hepatitis C virus (HCV) infection affects millions of people and leads to liver fibrosis, cirrhosis, and hepatocellular carcinoma. Treatment regimen selection requires HCV genotype (Gt) and Gt 1 subtype determination. Use of a laboratory developed, reverse transcription (RT)-PCR assay was explored as a low-cost, high-throughput screening approach for the major HCV genotypes and subtypes in North America. A commercial line probe assay (LiPA) was used for comparison. Sequencing and/or an alternative PCR assay were used for discordant analyses...
February 20, 2017: Journal of Virological Methods
https://www.readbyqxmd.com/read/28093275/reverse-transcription-loop-mediated-isothermal-amplification-for-rapid-detection-of-respiratory-syncytial-virus-directly-from-nasopharyngeal-swabs
#14
Johannes Hoos, Rebecca Marie Peters, Julia Tabatabai, Jürgen Grulich-Henn, Paul Schnitzler, Johannes Pfeil
Respiratory syncytial virus (RSV) is a common cause of lower respiratory tract infection in young infants and a major cause of nosocomial infection in pediatric care. Currently available RSV point-of-care tests are of limited sensitivity and relatively expensive. We developed and evaluated a novel RSV rapid test for use at point-of-care, based on reverse-transcription loop-mediated isothermal amplification (RT-LAMP) for direct testing of nasopharyngeal swab specimens. RT-LAMP can detect RSV within 30min, without the need for RNA extraction...
January 16, 2017: Journal of Virological Methods
https://www.readbyqxmd.com/read/28087375/super-oxidized-water-inactivates-major-viruses-circulating-in-swine-farms
#15
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
https://www.readbyqxmd.com/read/28082165/an-efficient-and-high-fidelity-method-for-amplification-cloning-and-sequencing-of-complete-tospovirus-genomic-rna-segments
#16
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
https://www.readbyqxmd.com/read/28082164/a-neutralization-assay-with-a-severe-fever-with-thrombocytopenia-syndrome-virus-strain-that-makes-plaques-in-inoculated-cells
#17
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
https://www.readbyqxmd.com/read/28069472/semi-quantitative-real-time-pcr-a-useful-approach-to-identify-persons-with-low-replicative-chronic-hepatitis-b
#18
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 2000IU/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 2000IU/ml was compared to a quantitative assay (Roche CobasAmpliPrep/CobasTaqMan HBV Test v2...
January 7, 2017: Journal of Virological Methods
https://www.readbyqxmd.com/read/28065747/microscopy-based-studies-on-the-interaction-of-bio-based-silver-nanoparticles-with-bombyx-mori-nuclear-polyhedrosis-virus
#19
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
https://www.readbyqxmd.com/read/28042018/pcr-activated-cell-sorting-as-a-general-cultivation-free-method-for-high-throughput-identification-and-enrichment-of-virus-hosts
#20
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
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