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Journal of Virological Methods

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https://www.readbyqxmd.com/read/28414163/droplet-digital-pcr-ddpcr-vs-quantitative-real-time-pcr-qpcr-approach-for-detection-and-quantification-of-merkel-cell-polyomavirus-mcpyv-dna-in-formalin-fixed-paraffin-embedded-ffpe-cutaneous-biopsies
#1
Rosaria Arvia, Mauro Sollai, Federica Pierucci, Carmelo Urso, Daniela Massi, Krystyna Zakrzewska
BACKGROUND: Merkel cell polyomavirus (MCPyV) is associated with Merkel cell carcinoma and high viral load in the skin was proposed as a risk factor for the occurrence of this tumour. MCPyV DNA was detected, with lower frequency, in different skin cancers but since the viral load was usually low, the real prevalence of viral DNA could be underestimated. OBJECTIVE: To evaluate the performance of two assays (qPCR and ddPCR) for MCPyV detection and quantification in formalin fixed paraffin embedded (FFPE) tissue samples...
April 14, 2017: Journal of Virological Methods
https://www.readbyqxmd.com/read/28411129/detection-of-a-novel-highly-pathogenic-h7-influenza-virus-by-duplex-real-time-reverse-transcription-polymerase-chain-reaction
#2
Weixin Jia, Chenfu Cao, Yanxing Lin, Liangning Zhong, Shumin Xie, Xiao Wang, Sanhong Yin, Zhenna Xu, Yixue Dai, Zhixian Li, Xiao Niu, Wenbao Qi, Tikang Lu, Ming Liao
No abstract text is available yet for this article.
April 11, 2017: Journal of Virological Methods
https://www.readbyqxmd.com/read/28408312/hcv-rna-quantification-in-liver-bioptic-samples-and-extrahepatic-compartments-using-the-abbott-realtime-hcv-assay
#3
Francesco Paolo Antonucci, Valeria Cento, Maria Chiara Sorbo, Matteo Ciancio Manuelli, Ilaria Lenci, Daniele Sforza, Domenico Di Carlo, Martina Milana, Tommaso Maria Manzia, Mario Angelico, Giuseppe Tisone, Carlo Federico Perno, Francesca Ceccherini-Silberstein
BACKGROUND & AIMS: We evaluated the performance of a rapid method to quantify HCV-RNA in the hepatic and extrahepatic compartments, by using for the first time the Abbott RealTime HCV-assay. METHODS: Non-tumoral (NT), tumoral (TT) liver samples, lymph nodes and ascitic fluid from patients undergoing orthotopic-liver-transplantation (N=18) or liver resection (N=4) were used for the HCV-RNA quantification; 5/22 patients were tested after or during direct acting antivirals (DAA) treatment...
April 11, 2017: Journal of Virological Methods
https://www.readbyqxmd.com/read/28392409/a-rapid-isothermal-assay-for-the-detection-of-hop-stunt-viroid-in-hop-plants-humulus-lupulus-and-its-application-in-disease-surveys
#4
Madhu Kappagantu, Dan Edward V Villamor, Jeff M Bullock, Kenneth C Eastwell
Hop stunt disease caused by Hop stunt viroid (HSVd) is a growing threat to hop cultivation globally. HSVd spreads mainly by use of contaminated planting material and by mechanical means. Thorough testing of hop yards and removal of infected bines are critical components of efforts to control the spread of the disease. Reverse transcription-polymerase chain reaction (RT-PCR) has become the primary technique used for HSVd detection; however, sample handling and analysis are technically challenging. In this study, a robust reverse transcription-recombinase polymerase amplification (RT-RPA) assay was developed to facilitate analysis of multiple samples...
April 7, 2017: Journal of Virological Methods
https://www.readbyqxmd.com/read/28365410/performance-evaluation-of-direct-fluorescent-antibody-focus-diagnostics-simplexa%C3%A2-flu-a-b-rsv-and-multi-parameter-customized-respiratory-taqman%C3%A2-array-card-in-immunocompromised-patients
#5
Deborah Steensels, Marijke Reynders, Patrick Descheemaeker, Martin D Curran, Frédérique Jacobs, Olivier Denis, Marie-Luce Delforge, Isabel Montesinos
BACKGROUND: Molecular assays for diagnosis of Flu A, Flu B, and RSV with short turn-around-time (TAT) are of considerable clinical importance. In addition, rapid and accurate diagnosis of a large panel of viral and atypical pathogens can be crucial in immunocompromised patients. OBJECTIVES: First, to evaluate the performance of the Simplexa™ Direct assay system in comparison with direct fluorescent antibody (DFA) and customized Taqman® Array Card (TAC) testing for RSV, Flu A, and Flu B in immunocompromised patients...
March 30, 2017: Journal of Virological Methods
https://www.readbyqxmd.com/read/28363451/characterization-of-three-commercial-elisa-kits-for-detection-of-bohv-1-ge-specific-antibodies-in-serum-and-milk-samples-and-applicability-of-bulk-milk-for-determination-of-herd-status
#6
Marylène Tignon, Miet De Baere, Jean-Baptiste Hanon, Annelies Goolaerts, Jean-Yves Houtain, Laurent Delooz, Ann Brigitte Cay
Vaccination of animals with gE-deleted vaccine strains (gE- marker vaccines) and differential detection of vaccinated vs infected animals with antibody ELISA targeting the gE or the gB proteins have been proved to be useful tools in programs for control and eradication of the bovine herpesvirus 1 (BoHV-1) responsible for infectious bovine rhinotracheitis (IBR), a major pathogen of cattle. The diagnostic sensitivity (DSe) and specificity (DSp) of three commercial gE ELISA kits from IDEXX, IDVet and CIV-HIPRA were compared for serum and milk matrices...
March 28, 2017: Journal of Virological Methods
https://www.readbyqxmd.com/read/28359920/hcv-core-antigen-as-an-alternate-test-to-hcv-rna-for-assessment-of-virologic-responses-to-all-oral-interferon-free-treatment-in-hcv-genotype-1-infected-patients
#7
Jürgen Kurt Rockstroh, Jordan J Feld, Stéphane Chevaliez, Kevin Cheng, Heiner Wedemeyer, Christoph Sarrazin, Benjamin Maasoumy, Christine Herman, John Hackett, Daniel E Cohen, George J Dawson, Gavin Cloherty, Jean-Michel Pawlotsky
In light of the advances in HCV therapy, simplification of diagnosis confirmation, pre- treatment diagnostic workup and treatment monitoring is required to ensure broad access to interferon-free therapies. HCV core antigen (HCV cAg) testing is rapid, giving results in approximately 60min, and less expensive than HCV RNA methods. While extensive data on the analytical performance of HCV cAg relative to RNA or comparisons in longitudinal studies of patients on interferon based (response guided) therapy there is very limited data on the relative performance of HCV cAg in diagnosis and monitoring patients receiving all-oral interferon free regimens...
March 27, 2017: Journal of Virological Methods
https://www.readbyqxmd.com/read/28359770/evaluation-of-cell-viability-dyes-in-antiviral-assays-with-rna-viruses-that-exhibit-different-cytopathogenic-properties
#8
Donald F Smee, Brett L Hurst, W Joseph Evans, Nathan Clyde, Sean Wright, Christopher Peterson, Kie-Hoon Jung, Craig W Day
Studies were conducted to determine the performance of four dyes in assessing antiviral activities of compounds against three RNA viruses with differing cytopathogenic properties. Dyes included alamarBlue(®) measured by absorbance (ALB-A) and fluorescence (ALB-F), neutral red (NR), Viral ToxGlo™ (VTG), and WST-1. Viruses were chikungunya, dengue type 2, and Junin, which generally cause 100, 80-90, and 50% maximal cytopathic effect (CPE), respectively, in Vero 76. Compounds evaluated were 6-azauridine, BCX-4430, 3-deazaguanine, EICAR, favipiravir, infergen, mycophenolic acid (MPA), ribavirin, and tiazofurin...
March 27, 2017: Journal of Virological Methods
https://www.readbyqxmd.com/read/28356221/rapid-detection-of-all-known-ebolavirus-species-by-reverse-transcription-loop-mediated-isothermal-amplification-rt-lamp
#9
Olamide K Oloniniyi, Yohei Kurosaki, Hiroko Miyamoto, Ayato Takada, Jiro Yasuda
Ebola virus disease (EVD), a highly virulent infectious disease caused by ebolaviruses, has a fatality rate of 25-90%. Without a licensed chemotherapeutic agent or vaccine for the treatment and prevention of EVD, control of outbreaks requires accurate and rapid diagnosis of cases. In this study, five sets of six oligonucleotide primers targeting the nucleoprotein gene were designed for specific identification of each of the five ebolavirus species using reverse transcription-loop mediated isothermal amplification (RT-LAMP) assay...
March 26, 2017: Journal of Virological Methods
https://www.readbyqxmd.com/read/28347708/analytical-validation-of-a-reverse-transcriptase-droplet-digital-pcr-rt-ddpcr-for-quantitative-detection-of-infectious-hematopoietic-necrosis-virus
#10
Peng Jia, Maureen K Purcell, Guang Pan, Jinjin Wang, Shifu Kan, Yin Liu, Xiaocong Zheng, Xiujie Shi, Junqiang He, Li Yu, Qunyi Hua, Tikang Lu, Wensheng Lan, James R Winton, Ningyi Jin, Hong Liu
Infectious hematopoietic necrosis virus (IHNV) is an important pathogen of salmonid fishes. A validated universal reverse transcriptase quantitative PCR (RT-qPCR) assay that can quantify levels of IHNV in fish tissues has been previously reported. In the present study, we adapted the published set of IHNV primers and probe for use in a reverse-transcriptase droplet digital PCR (RT-ddPCR) assay for quantification of the virus in fish tissue samples. The RT-ddPCR and RT-qPCR assays detected 13 phylogenetically diverse IHNV strains, but neither assay produced detectable amplification when RNA from other fish viruses was used...
March 24, 2017: Journal of Virological Methods
https://www.readbyqxmd.com/read/28342740/the-development-of-a-real-time-reverse-transcription-polymerase-chain-reaction-rrt-pcr-assay-using-taqman-technology-for-the-pan-detection-of-bluetongue-virus-btv
#11
Catherine Mulholland, Michael J McMenamy, Bernd Hoffmann, Bernadette Earley, Bryan Markey, Joseph Cassidy, Gordon Allan, Michael D Welsh, John McKillen
Bluetongue virus (BTV) is an infectious, non-contagious viral disease of domestic and wild ruminants that is transmitted by adult females of certain Culicoides species. Since 2006, several serotypes including BTV-1, 2, 4, 6, 8, 9 and 16, have spread from the Mediterranean basin into Northern Europe for the first time. BTV-8 in particular, caused a major epidemic in northern Europe. As a result, it is evident that most European countries are at risk of BTV infection. The objective of this study was to develop and validate a real-time reverse transcriptase-polymerase chain reaction (rRT-PCR) assay based on TaqMan technology for the detection of representative strains of all BTV serotypes...
March 23, 2017: Journal of Virological Methods
https://www.readbyqxmd.com/read/28336367/new-real-time-and-conventional-rt-pcrs-for-updated-molecular-diagnosis-of-infectious-bronchitis-virus-infection-ibv-in-chickens-in-egypt-associated-with-frequent-co-infections-with-avian-influenza-and-newcastle-disease-viruses
#12
Mahmoud M Naguib, Magdy F El-Kady, Dörte Lüschow, Kareem E Hassan, Abdel-Satar Arafa, Ali El-Zanaty, Mohamed K Hassan, Hafez M Hafez, Christian Grund, Timm C Harder
In Egypt, currently two geographically restricted genotypes of the infectious bronchitis coronavirus (IBV) are circulating with detrimental effects for poultry industry. A sensitive real-time RT-PCR assay targeting the IBV nucleocapsid gene (N) was developed to screen clinical samples for presence of IBV. Conventional RT-PCRs amplifying hypervariable regions (HVRs 1-2 and 3) of the IBV S1 gene were developed and amplificates used for nucleotide sequence-based typing of IBV field strains in Egyptian chickens directly from clinical samples...
March 21, 2017: Journal of Virological Methods
https://www.readbyqxmd.com/read/28315718/absolute-quantification-of-cassava-brown-streak-virus-mrna-by-real-time-qpcr
#13
Rudolph R Shirima, Daniel G 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 16, 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
#14
Gudrun S 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 16, 2017: Journal of Virological Methods
https://www.readbyqxmd.com/read/28315379/magnetic-nanoparticles-for-efficient-cell-transduction-with-semliki-forest-virus
#15
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 15, 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
#16
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 12, 2017: Journal of Virological Methods
https://www.readbyqxmd.com/read/28257802/assessment-of-influenza-a-neuraminidase-subtype-n1-potency-by-elisa
#17
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
#18
Haroldo Cid da Silva Junior, 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 9, 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
#19
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
#20
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
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