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Journal of Virological Methods

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https://www.readbyqxmd.com/read/28923315/development-of-a-digital-droplet-pcr-assay-to-measure-hbv-dna-in-patients-receiving-long-term-tdf-treatment
#1
Yang Liu, Andrea L Cathcart, William E Delaney, Kathryn M Kitrinos
The COBAS TaqMan assay has a lower limit of quantification (LLOQ) of 169 HBV copies/mL and a lower limit of detection (LLOD) of 58 copies/mL. HBV DNA below the TaqMan LLOQ is classified as target not detected (TND) (<58 copies/mL) or target detected (TD) (between 58 to 169 copies/mL). Here we have developed a more sensitive digital droplet PCR (ddPCR) assay to evaluate the impact of long-term tenofovir disoproxil fumarate (TDF) treatment in patients that did or did not achieve HBsAg seroconversion. A ddPCR assay was developed to detect HBV DNA to 8 copies/mL...
September 15, 2017: Journal of Virological Methods
https://www.readbyqxmd.com/read/28923314/a-simple-method-for-isolation-of-cell-associated-viral-particles-from-cell-culture
#2
Katarina Laposova, Ingrid Oveckova, Jana Tomaskova
A common method for cell-associated virus isolation involves disruption of infected cells by a combination of hypotonic burst, freeze-thaw cycles (F-T) and sonication. This protocol was also originally used for the preparation of cell-free extract containing the MX strain of lymphocytic choriomeningitis virus (LCMV), which is preferentially propagated by cell-to-cell contact and does not release distinct virions into the medium. In the present study, we compared different approaches to virus isolation. Based on virus yield, we show that deionized water lysis is the fastest and most effective method for releasing LCMV MX infectious viral particles from persistently infected cells...
September 15, 2017: Journal of Virological Methods
https://www.readbyqxmd.com/read/28919035/development-of-a-fast-and-efficient-method-for-hepatitis-a-virus-concentration-from-green-onion
#3
Yan Zheng, Yuan Hu
Hepatitis A virus (HAV) can cause serious liver disease and even death. HAV outbreaks are associated with the consumption of raw or minimally processed produce, making it a major public health concern. Infections have occurred despite the fact that effective HAV vaccine has been available. Development of a rapid and sensitive HAV detection method is necessary for an investigation of an HAV outbreak. Detection of HAV is complicated by the lack of a reliable culture method. In addition, due to the low infectious dose of HAV, these methods must be very sensitive...
September 14, 2017: Journal of Virological Methods
https://www.readbyqxmd.com/read/28919034/a-sensitive-and-specific-lateral-flow-assay-for-rapid-detection-of-antibodies-against-glycoprotein-b-of-aujeszky-s-disease-virus
#4
Veronika V Vrublevskaya, Vladimir N Afanasyev, Andrey A Grinevich, Yuri Y Skarga, Pavel P Gladyshev, Sagila A Ibragimova, Dmitry V Krylsky, Sergey V Dezhurov, Oleg S Morenkov
A direct double antibody lateral flow assay (DDA-gB-LFA) for the detection of antibodies against the glycoprotein B (gB) of Aujeszky's disease virus (ADV) in swine sera was developed. A native ADV gB was used for the preparation of a conjugate with colloidal gold particles and the immobilization on the strip membrane. The gB purified from ADV virions by immunoaffinity chromatography retained its native epitope structure after adsorption on the nitrocellulose membrane and the surface of colloidal gold particles...
September 14, 2017: Journal of Virological Methods
https://www.readbyqxmd.com/read/28918076/simultaneous-detection-of-wheat-dwarf-virus-northern-cereal-mosaic-virus-barley-yellow-striate-mosaic-virus-and-rice-black-streaked-dwarf-virus-in-wheat-by-multiplex-rt-pcr
#5
Peipei Zhang, Yan Liu, Wenwen Liu, Sebastien Massart, Xifeng Wang
Wheat dwarf virus (WDV), barley yellow striate mosaic virus (BYSMV), rice black-streaked dwarf virus (RBSDV) and northern cereal mosaic virus (NCMV) are four viruses infecting wheat and causing similar symptoms. In this paper, a multiplex reverse transcription polymerase chain reaction (m-RT-PCR) method has been developed for the simultaneous detection and discrimination of these viruses. The protocol uses specific primer set for each virus and produces four distinct fragments (273, 565, 783 and 1296bp), detecting the presence of RBSDV, BYSMV, WDV and NCMV, respectively...
September 13, 2017: Journal of Virological Methods
https://www.readbyqxmd.com/read/28918075/direct-detection-of-trichomonas-vaginalis-virus-in-trichomonas-vaginalis-positive-clinical-samples-from-the-netherlands
#6
Ivo Jehee, Charlotte van der Veer, Michelle Himschoot, Mirjam Hermans, Sylvia Bruisten
Trichomonas vaginalis is the most common sexually transmitted parasitical infection worldwide. T. vaginalis can carry a virus: Trichomonas vaginalis virus (TVV). To date, four TVV species have been described. Few studies have investigated TVV prevalence and its clinical importance. We have developed a nested reverse-transcriptase PCR, with novel, type specific primers to directly detect TVV RNA in T. vaginalis positive clinical samples. A total of 119T. vaginalis positive clinical samples were collected in Amsterdam and "s-Hertogenbosch, the Netherlands, from 2012 to 2016...
September 13, 2017: Journal of Virological Methods
https://www.readbyqxmd.com/read/28918074/rapid-adenovirus-typing-method-for-species-identification
#7
Fabienne Rayne, Linda Wittkop, Clément Bader, Somar Kassab, Camille Tumiotto, Sylvie Berciaud, Harald Wodrich, Marie-Edith Lafon
Adenoviruses are characterized by a large variability, reflected by their classification in species A to G. Certain species, eg A and C, could be associated with increased clinical severity, both in immunocompetent and immunocompromised hosts suggesting that in some instances species identification provides clinically relevant information. Here we designed a novel "pVI rapid typing method" to obtain quick, simple and cost effective species assignment for Adenoviruses, thanks to combined fusion temperature (Tm) and amplicon size analysis...
September 13, 2017: Journal of Virological Methods
https://www.readbyqxmd.com/read/28918073/a-directed-evolution-approach-to-select-for-novel-adeno-associated-virus-capsids-on-an-hiv-1-producer-t-cell-line
#8
Dawn P Wooley, Priyanka Sharma, John R Weinstein, Poornima Kotha Lakshmi Narayan, David V Schaffer, Katherine J D A Excoffon
A directed evolution approach was used to select for Adeno-associated virus (AAV) capsids that would exhibit more tropism toward an HIV-1 producer T cell line with the long-term goal of developing improved gene transfer vectors. A library of AAV variants was used to infect H9T cells previously infected or uninfected by HIV-1 followed by AAV amplification with wild-type adenovirus. Six rounds of biological selection were performed, including negative selection and diversification after round three. The H9T cells were successfully infected with all three wild-type viruses (AAV, adenovirus, and HIV-1)...
September 13, 2017: Journal of Virological Methods
https://www.readbyqxmd.com/read/28918072/development-of-a-duplex-semi-nested-pcr-assay-for-detection-of-classical-goose-parvovirus-and-novel-goose-parvovirus-related-virus-in-sick-or-dead-ducks-with-short-beak-and-dwarfism-syndrome
#9
Pengfei Li, Ruihua Zhang, Junhao Chen, Dapeng Sun, Jingjing Lan, Shaoli Lin, Shasha Song, Zhijing Xie, Shijin Jiang
Duck short beak and dwarfism syndrome (SBDS) is an emerging infectious disease caused by a novel goose parvovirus-related virus (NGPV) in China. Until now, it remains uncertain whether the Cherry Valley ducks and mule ducks with SBDS are co-infected with classical goose parvovirus (GPV) and NGPV. In this study, a duplex semi-nested PCR assay with high specificity and sensitivity was developed for detection of the two viruses. Using the duplex PCR assay, NGPV was tested positive in all the 15 duck flocks with SBDS, whereas classical GPV was not detected in all the 133 sick and dead ducks collected from East China...
September 13, 2017: Journal of Virological Methods
https://www.readbyqxmd.com/read/28916427/simultaneous-detection-and-differentiation-of-three-potyviridae-viruses-in-sweet-potato-by-a-multiplex-taqman-real-time-rt-pcr-assay
#10
Pingxiu Lan, Fan Li, Jorge Abad, Lingling Pu, Ruhui Li
A multiplex TaqMan real time RT-PCR was developed for detection and differentiation of Sweet potato virus G, Sweet potato latent virus and Sweet potato mild mottle virus in one tube. Amplification and detection of a fluorogenic cytochrome oxidase gene was included as an internal control. The assay was compared with a multiplex RT-PCR developed in the initial study for the detection and differentiation of the three viruses and host 18S rRNA. Primers and/or probes of the two assays were designed from conserved regions of each virus...
September 12, 2017: Journal of Virological Methods
https://www.readbyqxmd.com/read/28916419/development-of-an-indirect-enzyme-linked-immunosorbent-assay-elisa-to-differentiate-antibodies-against-wild-type-porcine-reproductive-and-respiratory-syndrome-from-the-vaccine-strain-tjm-f92-based-on-a-recombinant-nsp2-protein
#11
X X Wang, F X Wang, Z G Li, Y J Wen, X Wang, N Song, H Wu
An accurate ELISA method to differentiate pigs infected with wild-type porcine reproductive and respiratory syndrome (PRRSV) strains from vaccinated ones would help to monitor PRRSV vaccination compliance. The recombinant protein GST-d120aa derived from the continuous deletion of 120 amino acids in the non-structural protein 2 region of the modified-live vaccine strain TJM-F92 was used to develop an indirect enzyme-linked immunosorbent assay (d120-ELISA) for differentiating serum antibodies against TJM-F92 from other PRRSV strains...
September 12, 2017: Journal of Virological Methods
https://www.readbyqxmd.com/read/28893550/specificity-of-two-hiv-screening-tests-detecting-simultaneously-hiv-1-p24-antigen-and-antibodies-to-hiv-1-and-2
#12
Annette Blaich, Andreas Buser, Marcel Stöckle, Christian Gehringer, Hans H Hirsch, Manuel Battegay, Thomas Klimkait, Reno Frei
This study aimed at assessing the specificity of the Elecsys(®) HIV combi PT in comparison to the ARCHITECT(®) HIV Ag/Ab Combo. With both of these assays, 3997 unselected sera from patients of a tertiary health care centre in Basel, Switzerland, were screened for HIV. Reactive sera were reanalysed on the VIDAS(®) HIV Duo Ultra to identify false-reactive specimens prior to confirmation by quantitative PCR and line immunoassay. The Elecsys(®) compared to the ARCHITECT(®) shows a similar specificity (99.7% versus 99...
September 8, 2017: Journal of Virological Methods
https://www.readbyqxmd.com/read/28888402/investigating-the-potential-of-multiwalled-carbon-nanotubes-based-zinc-nanocomposite-as-a-recognition-interface-towards-plant-pathogen-detection
#13
Muhammad Ali Tahir, Sadaf Hameed, Anam Munawar, Imran Amin, Shahid Mansoor, Waheed S Khan, Sadia Zafar Bajwa
The emergence of nanotechnology has opened new horizons for constructing efficient recognition interfaces. This is the first report where the potential of a multiwalled carbon nanotube based zinc nanocomposite (MWCNTs-Zn NPs) investigated for the detection of an agricultural pathogen i.e. Chili leaf curl betasatellite (ChLCB). Atomic force microscope analyses revealed the presence of multiwalled carbon nanotubes (MWCNTs) having a diameter of 50-100nm with zinc nanoparticles (Zn-NPs) of 25-500nm. In this system, these bunches of Zn-NPs anchored along the whole lengths of MWCNTs were used for the immobilization of probe DNA strands...
September 6, 2017: Journal of Virological Methods
https://www.readbyqxmd.com/read/28887190/rapid-detection-of-fifteen-known-soybean-viruses-by-dot-immunobinding-assay
#14
Akhtar Ali
A dot-immunobinding assay (DIBA) was optimized and used successfully for the rapid detection of 15 known viruses [Alfalfa mosaic virus (AMV), Bean pod mottle virus (BPMV), Bean yellow mosaic virus (BYMV), Cowpea mild mottle virus (CPMMV), Cowpea severe mosaic virus (CPSMV), Cucumber mosaic virus (CMV), Peanut mottle virus (PeMoV), Peanut stunt virus (PSV), Southern bean mosaic virus (SBMV), Soybean dwarf virus (SbDV), Soybean mosaic virus (SMV), Soybean vein necrosis virus (SVNV), Tobacco ringspot virus (TRSV), Tomato ringspot virus (ToRSV), and Tobacco streak virus (TSV)] infecting soybean plants in Oklahoma...
September 6, 2017: Journal of Virological Methods
https://www.readbyqxmd.com/read/28882762/estimation-of-the-diagnostic-performance-of-two-elisas-to-detect-pcv2-antibodies-in-pig-sera-using-a-bayesian-method
#15
C Fablet, N Rose, C Bernard, I Messager, Y Piel, B Grasland
This study was designed to assess the diagnostic characteristics of two PCV2 ELISAs without a gold standard. Four hundred and sixty-five serum samples from finishing pigs (25 herds) not vaccinated against PCV2 were used. Samples were tested by two ELISAs: an in-house ELISA (I-ELISA) and the commercial SERELISA(®)PCV2 Ab Mono Blocking kit (S-ELISA). A ROC curve was used to assess the S-ELISA's optimal threshold by taking the I-ELISA as a reference and using the cut-off previously determined by comparison to an cccmonolayer assay (IPMA)...
September 5, 2017: Journal of Virological Methods
https://www.readbyqxmd.com/read/28855093/metagenomic-arbovirus-detection-using-minion-nanopore-sequencing
#16
Jana Batovska, Stacey E Lynch, Brendan C Rodoni, Tim I Sawbridge, Noel Oi Cogan
With its small size and low cost, the hand-held MinION sequencer is a powerful tool for in-field surveillance. Using a metagenomic approach, it allows non-targeted detection of viruses in a sample within a few hours. This study aimed to determine the ability of the MinION to metagenomically detect and characterise a virus from an infected mosquito. RNA was extracted from an Aedes notoscriptus mosquito infected with Ross River virus (RRV), converted into cDNA and sequenced on the MinION. Bioinformatic analysis of the MinION reads led to detection of full-length RRV, with reads of up to 2...
September 4, 2017: Journal of Virological Methods
https://www.readbyqxmd.com/read/28867302/comparison-of-the-efficiency-of-different-newcastle-disease-virus-reverse-genetics-systems
#17
Haijin Liu, Renata Servan de Almeida, Patricia Gil, Emmanuel Albina
Rescue of negative-sense single-stranded RNA viruses ((-)ssRNA virus), generally requires the handling of a large number of plasmids to provide the virus genome and essential components for gene expression and genome replication. This constraint probably renders reverse genetics of (-)ssRNA virus more complex and less efficient. Some authors have shown that the fewer the plasmids, the more efficient reverse genetics is for segmented RNA virus. However, it is not clear if the same applies for (-)ssRNA, such as Newcastle disease virus (NDV)...
September 1, 2017: Journal of Virological Methods
https://www.readbyqxmd.com/read/28866364/development-of-rapid-and-highly-sensitive-detection-of-bean-common-mosaic-necrosis-virus-in-leguminous-crops-using-loop-mediated-isothermal-amplification-assay
#18
Siwon Lee, Heejung Kim, Jin-Young Lee, Jae-Young Rho
Bean common mosaic necrosis virus (BCMNV) is a plant pathogenic virus that can infect leguminous crops such as kidney beans, sunn hemp, red beans, and mung beans. BCMNV has not been reported in Korea and is classified as a quarantine plant virus. Currently, the standard diagnostic method for diagnosis of BCMNV is reverse transcription (RT)-nested PCR system. However a more rapid monitoring system is needed to enable the testing of more samples. The use of highly efficient loop-mediated isothermal amplification (LAMP) assay for its detection has not yet been reported, and development of LAMP for detecting BCMNV in this study...
September 1, 2017: Journal of Virological Methods
https://www.readbyqxmd.com/read/28864043/development-of-an-immunochromatographic-strip-for-detection-of-antibodies-against-duck-tembusu-virus
#19
Jie Deng, Yanhong Liu, Renyong Jia, Mingshu Wang, Shun Chen, Dekang Zhu, Mafeng Liu, Kunfeng Sun, Xinxin Zhao, Zhongqiong Yin, Anchun Chen
Duck Tembusu virus (DTMUV), a novel flavivirus, causes severe disease in ducks. There is an urgent need for a rapid and effective diagnostic method to control the spread of DTMUV. We chose the envelope (E) protein from DTMUV as an antigen and combined it with colloidal gold particles as tracers to specifically detect anti-DTMUV antibodies. Based on the double-antigen sandwich format, an immunochromatographic strip (ICS) for the rapid detection of anti-DTMUV antibodies was developed. The ICS showed a high specificity and no cross-reactivity with other sera...
August 31, 2017: Journal of Virological Methods
https://www.readbyqxmd.com/read/28837842/defining-the-relative-performance-of-isothermal-assays-that-can-be-used-for-rapid-and-sensitive-detection-of-foot-and-mouth-disease-virus
#20
Emma L A Howson, Yohei Kurosaki, Jiro Yasuda, Masayoshi Takahashi, Hiroaki Goto, Ashley R Gray, Valerie Mioulet, Donald P King, Veronica L Fowler
This study describes the first multiway comparison of portable isothermal assays for the detection of foot-and-mouth disease virus (FMDV), benchmarked against real-time reverse transcription RT-PCR (rRT-PCR). The selected isothermal chemistries included reverse transcription loop-mediated isothermal amplification (RT-LAMP) and reverse transcription recombinase polymerase amplification (RT-RPA). The analytical sensitivity of RT-LAMP was comparable to rRT-PCR (10(1) RNA copies), while RT-RPA was one log10 less sensitive (10(2) RNA copies)...
August 31, 2017: Journal of Virological Methods
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