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Journal of Virological Methods

Jianchang Wang, Ruoxi Zhang, Jinfeng Wang, Qingan Han, Libing Liu, Yanan Li, Wanzhe Yuan
Porcine epidemic diarrhea (PED), which is caused by porcine epidemic diarrhea virus (PEDV), is an acute, highly contagious enteric disease characterized by severe watery diarrhea, vomiting, dehydration, and high mortality in suckling piglets. A real-time reverse-transcription recombinase polymerase amplification assay (RT-RPA) was developed based on the nucleocapsid gene of PEDV. RT-RPA assay was performed at 40 °C for 20 min. The assay could detect both the classical and variant PEDV strains, and there was no cross-reaction with other pathogens tested...
January 9, 2018: Journal of Virological Methods
Yogita Maheshwari, Vijayanandraj Selvaraj, Rakesh Kumar Jain, Bikash Mandal
A simple and rapid lateral flow immunoassay (LFIA) was developed by utilizing gold nanoparticles conjugated to a polyclonal antibody against coat protein of large cardamom chirke virus (LCCV). The LFIA was based on the principle of sandwich immunoassay, detected LCCV within ∼10 minutes and the results could be evaluated visually. The colloidal gold (CG) was made using 1% gold chloride solution. The LCCV IgG (1 μg/μl) and Mouse IgG (0.5 μg/μl) were conjugated with CG individually and coated onto a conjugate pad at 1:1 ratio...
December 27, 2017: Journal of Virological Methods
Wenzhi Liu, Yong Zhou, Yuding Fan, Nan Jiang, Kenneth Cain, Lingbing Zeng
Infectious spleen and kidney necrosis virus (ISKNV) has been recognized as the causative agent of the most serious disease in cultured mandarin fish, Siniperca chuatsi, in China. Disease outbreaks have resulted in substantial losses to the aquaculture industry. Currently, reliable laboratory detection and identification methods are available for this virus. However, rapid detection methods applicable for on-site diagnosis of this infectious agent are unavailable. To address this need, a nearly instrument-free, cost-effective and simple detection method was developed and optimized and incorporates cross priming amplification coupled with vertical flow visualization for rapid identification of ISKNV (ISKNV-CPA-VF)...
December 26, 2017: Journal of Virological Methods
Phuc H Pham, Bibi S H Sokeechand, Kyle A Garver, Ginny Jones, John S Lumsden, Niels C Bols
RNAlater is a commonly used transport and storage solution for samples collected for fish health investigations, particularly those potentially involving viruses. However, the infectivity of fish viruses after storage in RNAlater have not been determined. Nevertheless, knowledge of pathogen infectivity of preserved samples is crucial for ensuring safe transport and storage protocols. Therefore, the infectivity of three fish RNA viruses in RNAlater was examined at four temperatures: -80 °C, 4 °C, room temperature (RT, approximately 22 °C) and 37 °C...
December 26, 2017: Journal of Virological Methods
Yu-Ri Park, Hye-Ryung Kim, Seong-Hee Kim, Kyoung-Ki Lee, Young S Lyoo, Sang-Geon Yeo, Choi-Kyu Park
A loop-mediated isothermal amplification (LAMP) assay using hydroxynaphthol blue was developed for the rapid and visual detection of the capsid gene of porcine circovirus 3 (PCV3). The amplification could be completed in 40 min at 62 °C, and the results could be visually detected by the naked eye. The assay specifically amplified PCV3 DNA and not other porcine viral nucleic acids. The limit of detection of the assay was 50 PCV3 DNA copies, which was comparable to that of the real-time polymerase chain reaction (qPCR) and lower than that of conventional PCR...
December 22, 2017: Journal of Virological Methods
Vikrant Sharma, Dhruva Chaudhry, Samander Kaushik
BACKGROUND: Influenza A viruses (IAVs) have always remain a serious concern for the global economic and public health. A rapid, specific and sensitive detection method is always needed to control the influenza in its early stages by timely intervention of therapy and early clinical management. OBJECTIVES: To develop RT-LAMP assays for detection of influenza A viruses, their further subtyping into seasonal (H1N1, H3N2) and novel pandemic H1N1 viruses and to evaluate clinical applicability of optimized RT-LAMP assays on patients' samples...
December 15, 2017: Journal of Virological Methods
Aayushi Uberoi, Satoshi Yoshida, Paul F Lambert
Preclinical model systems to study multiple features of the papillomavirus life cycle are extremely valuable tools to aid our understanding of Human Papillomavirus (HPV) biology, disease progression and treatments. Mouse papillomavirus (MmuPV1) is the first ever rodent papillomavirus that can infect the laboratory strain of mice and was discovered recently in 2011. This model is an attractive model to study papillomavirus pathogenesis due to the ubiquitous availability of lab mice and the fact that this mouse species is easily genetically modifiable...
December 15, 2017: Journal of Virological Methods
Amanda Poe, Ngocvien Thi Duong, Kanwar Bedi, Maja Kodani
Diagnosis of hepatitis C virus (HCV) infection is based on testing for antibodies to HCV (anti-HCV), hepatitis C core antigen (HCV cAg) and HCV RNA. To ensure quality control (QC) and quality assurance (QA), proficiency panels are provided by reference laboratories and various international organizations, requiring costly dry ice shipments to maintain specimen integrity. Alternative methods of specimen preservation and transport can save on shipping and handling and help in improving diagnostics by facilitating QA/QC of various laboratories especially in resource limited countries...
December 14, 2017: Journal of Virological Methods
Zahra Rikhtegaran Tehrani, Kayhan Azadmanesh, Ehsan Mostafavi, Safoora Gharibzadeh, Shahrzad Soori, Mohammad Azizi, Alireza Khabiri
Estimation of HIV incidence provides real-time information of HIV transmission trends for decision makers. Anti-integrase antibodies are the last ones produced during seroconversion and presence of high-avidity anti-integrase antibodies indicates the chronicity of HIV infection. This study aimed to evaluate the performance of these antibodies in discriminating of recent from non-recent HIV infection. For this purpose, different ELISA formats were developed to detect high-avidity anti-integrase antibodies in a commercially available performance panel, and the best assay was selected for further evaluation...
December 14, 2017: Journal of Virological Methods
Leman Robin, Ralph-Sydney Mboumba Bouassa, Zita Aleyo Nodjikouambaye, Laura Charmant, Mathieu Matta, Sylvie Simon, Mounir Filali, Souleymane Mboup, Laurent Bélec
BACKGROUND: The HIV/HCV/HBsAg Triplex consists in manually performed, visually interpreted, lateral flow, immunochromatographic rapid diagnostic test simultaneously detecting in 15min human immunodeficiency virus (HIV)-1 and HIV-2 and hepatitis C virus (HCV)- specific antibodies (Ab) (IgG and IgM) and hepatitis B virus (HBV) surface antigen (HBsAg) in serum, plasma and whole blood. METHODS: A hospital-based cross-sectional study was conducted on a prospective panel of serum samples from adult inpatients included from routine analysis irrespectively of age and sex, including 250 sera positive for HIV-1-specific Ab, 250 for HCV-specific Ab, 250 for HBsAg and 250 sera negative for HIV- and HCV- Ab and HBsAg, and from 110 HIV-2-infected patients living in Ivory Coast, according to the results obtained by the reference chemiluminiscent microparticle immunoassay (CMIA) Abbott Architect i2000SR analyzer (Abbott Diagnostic, Chicago, IL, USA)...
December 5, 2017: Journal of Virological Methods
Xiujuan Peng, Alex Nguyen, Debadyuti Ghosh
TaqMan and SYBR Green quantitative PCR (qPCR) methods were developed as DNA-based approaches to reproducibly enumerate M13 and T7 phages from phage display selection experiments individually and simultaneously. The genome copies of M13 and T7 phages were quantified by TaqMan or SYBR Green qPCR referenced against M13 and T7 DNA standard curves of known concentrations. TaqMan qPCR was capable of quantifying M13 and T7 phage DNA simultaneously with a detection range of 2.75*101 - 2.75*108 genome copies(gc)/μL and 2...
November 28, 2017: Journal of Virological Methods
Daiki Kanbayashi, Takako Kurata, Kazuo Takahashi, Tetsuo Kase, Jun Komano
A large rubella outbreak occurred in Japan 2013, and 14,344 rubella and 45 congenital rubella syndrome (CRS) cases were reported. At that time, the populational immunity was above the protective threshold assessed by hemmaglutination inhibition (HI) titer. The genotype 2B rubella virus (RV) strains were responsible for the outbreak, which are non-indigenous in Japan. In this work, a cell-based high throughput assay was established to measure the neutralizing antibody (NA) titer against circulating RV isolates...
November 27, 2017: Journal of Virological Methods
Yongcan Lu, Bingyu Yao, Guoping Wang, Ni Hong
A reverse transcription loop-mediated isothermal amplification (RT-LAMP) assay was developed for the detection of Apple chlorotic leaf spot virus (ACLSV) and Apple stem pitting virus (ASPV), two important viruses frequently occurring in pear trees. A set of four RT-LAMP primers designed based on the highly conserved region of each CP gene of the two viruses showed high specificity and feasibility for ACLSV and ASPV detections. The RT-LAMP assays for ACLSV and ASPV in pear samples were 104 and 103 times more sensitive than that of conventional RT-PCR assays...
November 24, 2017: Journal of Virological Methods
Toshikatsu Shibata, Kuniaki Nerome, Mitsuhiko Moriyama, Satoshi Hayakawa, Kazumichi Kuroda
A previous report demonstrated that influenza virus infection induces accumulation of EGFP-tagged M1 protein (EGFP-M1) in the sub-nuclear domain ND10. Here, we show that the transfection of four viral protein (NP, PB2, PB1, PA) expression vectors and eight RNA segment expression vectors induced the formation of nuclear dots of EGFP-M1 as seen in virus infections. Omission of the segment 7 RNA expression vector, however, abolished the nuclear dots of EGFP-M1. This result suggests an essential role for authentic M1 protein and/or M2 protein, both of which are encoded in segment 7, in the formation of nuclear dots of EGFP-M1...
November 23, 2017: Journal of Virological Methods
M Biava, F Colavita, A Marzorati, D Russo, D Pirola, A Cocci, A Petrocelli, M Guanti Delli, G Cataldi, T A Kamara, A S Kamara, K Konneh, A Cannas, S Coen, S Quartu, S Meschi, M B Valli, A Mazzarelli, C Venditti, G Grassi, G Rozera, C Castilletti, A Mirazimi, M R Capobianchi, G Ippolito, R Miccio, A Di Caro
BACKGROUND: The 2013-2016 Ebola virus disease (EVD) outbreak showed a lack of diagnostic point-of-care methods. Currently, EBOV diagnosis relies on quantitative reverse-transcription-PCR (RT- qPCR), highly specific and sensitive, but requiring skilled personnel and well-equipped laboratories. In field settings, these factors and others, such as samples' time of collection and transportation, determine a prolonged turnaround-time to final results. In outbreak scenarios, a rapid and transportable method could eliminate issues of cohorting suspected and actual EVD patients for lack of diagnostic certainty...
November 20, 2017: Journal of Virological Methods
M J Ankcorn, S Ijaz, B Haywood, J Neuberger, A M Elsharkawy, J Maggs, R S Tedder
Genotype 3 hepatitis E virus (HEV) can lead to persistent infections in immunocompromised hosts. A recently available commercial assay for the detection of HEV antigen (HEV-Ag ELISA, Wantai diagnostics) may enable the study of HEV-Ag dynamics in such persistent infections, however currently there is no confirmatory test available. We generated a putative neutralising reagent from a pool of four convalescent blood donor samples and explored neutralising activity against HEV antigens from clinical samples, HEV tissue-culture and virus-like particles...
November 20, 2017: Journal of Virological Methods
Viska I Iskandar, Yutaka Sasaki, Naoto Yoshino, Raden Z R Abubakar, Shigehiro Sato, Yasushi Muraki
A cell-based vaccine production method for influenza virus may be an effective and more rapid alternative to egg-based systems. For high-yield virus production, the effect of bovine, porcine, fungal, and recombinant trypsins on influenza A/H1N1 virus replication in MDCK SI-6 cells (SI-6 cells), a novel MDCK cell line developed by our research group, was examined. SI-6 cells infected with influenza A/H1N1 virus were incubated in the presence of four trypsin types at various concentrations, and virus yields in the culture medium were evaluated by a hemagglutination (HA) assay...
November 16, 2017: Journal of Virological Methods
Cédric Charretier, Aure Saulnier, Loïc Benair, Corinne Armanet, Isabelle Bassard, Sandra Daulon, Bertrand Bernigaud, Emanuel Rodrigues de Sousa, Clémence Gonthier, Edouard Zorn, Emmanuelle Vetter, Claire Saintpierre, Patrice Riou, David Gaillac
The classical cell-culture methods, such as cell culture infectious dose 50% (CCID50) assays, are time-consuming, end-point assays currently used during the development of a viral vaccine production process to measure viral infectious titers. However, they are not suitable for handling the large number of tests required for high-throughput and large-scale screening analyses. Impedance-based bio-sensing techniques used in real-time cell analysis (RTCA) to assess cell layer biological status in vitro, provide real-time data...
November 14, 2017: Journal of Virological Methods
Michelle Wille, Hong Yin, Åke Lundkvist, Juan Xu, Shaman Muradrasoli, Josef D Järhult
Surveillance of wild birds is critical in monitoring for highly pathogenic avian influenza A viruses (AIVs). However, a successful surveillance regime requires proper treatment of samples in the field - rapid placement of samples in -80°C and subsequent maintenance of cold-chain. Given the logistical difficulties of this, many avian taxa and/or geographic locations are not sampled, or, when sampled may result in false negatives due to poor sample treatment in the field. Here, we assessed the utility of RNAlater(®) as a stabilization agent for AIV sampling...
November 10, 2017: Journal of Virological Methods
Junli Liu, Lu Yao, Feifei Zhai, Yuqing Chen, Jing Lei, Zhenwei Bi, Jianhua Hu, Qian Xiao, Suquan Song, Liping Yan, Jiyong Zhou
Avian influenza virus (AIV), especially subtypes H5, H7 and H9, has contributed to enormous economic losses and poses a potential pandemic threat to global human public health. Early screening of suspected cases is key to controlling the spread of AIVs. In this study, an accurate, rapid, and triplex real-time polymerase chain reaction (PCR) assay was developed for the simultaneous detection of AIV subtypes H5, H7 and H9. The sensitivity of the real-time PCR was at least 100 times higher than that of the conventional PCR, with a detection limit of 50 copies and an EID50 of 1 (50% egg infections dose) for the H5, H7, and H9 subtypes...
November 10, 2017: Journal of Virological Methods
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