journal
https://read.qxmd.com/read/38309371/diagnostic-performance-of-cross-priming-amplification-based-lateral-flow-assay-cpa-lfa-and-real-time-pcr-for-koi-herpesvirus-khv-detection
#21
JOURNAL ARTICLE
Guk Hyun Kim, Ye Jin Jeong, Yu Gyeong Jeon, Yun Jung Yang, Joon Gyu Min, Do Hyung Kim, Kwang Il Kim
Epizootics of Koi herpesvirus (KHV) cause mass mortality in koi carp (Cyprinus rubrofuscus) and common carp (Cyprinus carpio) worldwide. Rapid and accurate virus detection technology is crucial for preventing pathogen spread and minimizing damage. Although several diagnostic assays have been developed for KHV, the analytical and diagnostic performance of the detection methods has not been evaluated. In this study, we developed and validated the diagnostic performance of two molecular diagnostic assays, cross-priming amplification-based lateral flow assay (CPA-LFA) and TaqMan probe-based real-time polymerase chain reaction (PCR)...
February 1, 2024: Journal of Virological Methods
https://read.qxmd.com/read/38290650/visual-rapid-and-cost-effective-bk-virus-detection-system-for-renal-transplanted-patients-using-gold-nanoparticle-coupled-loop-mediated-isothermal-amplification-nanolamp
#22
JOURNAL ARTICLE
Sunil Kumar, Srishty Raman, Kishore Sesham, Abhishek Gupta, Raj Kanwar Yadav, Asit Ranjan Mridha, Subhash Chandra Yadav
A substantial percentage of kidney transplant recipients show transplant failure due to BK virus-induced nephropathy. This can be clinically controlled by the rapid and timely detection of BK virus infection in immune-compromised patients. We report a rapid (two hours from sample collection, processing, and detection), cost-effective (< 2$), highly sensitive and BKV-specific nanoLAMP (loop-mediated isothermal amplification) diagnostic methodology using novel primers and gold nanoparticles complex-based visual detection...
January 28, 2024: Journal of Virological Methods
https://read.qxmd.com/read/38246565/genome-sequencing-of-the-mpox-virus-2022-outbreak-with-amplicon-based-oxford-nanopore-minion-sequencing
#23
JOURNAL ARTICLE
Annika Brinkmann, Katharina Pape, Steven Uddin, Niklas Woelk, Sophie Förster, Heiko Jessen, Janine Michel, Claudia Kohl, Lars Schaade, Andreas Nitsche
We present an amplicon-based assay for MinION Nanopore sequencing of mpox virus (MPXV) genomes from clinical specimens, obtaining high-quality results with an average genome coverage of 99% for Ct values of up to 25, and a genome coverage of 97.1% for Ct values from 25 to 30 which are challenging to sequence. This assay is easy to implement in PCR-based workflows and provides accurate genomic data within a short time.
January 19, 2024: Journal of Virological Methods
https://read.qxmd.com/read/38246564/specific-antibody-production-using-recombinant-proteins-to-elucidate-seed-transmission-and-nuclear-localization-of-coguvirus-citrulli-and-coguvirus-henanense-in-radicles-of-watermelon-crop
#24
JOURNAL ARTICLE
Caterynne M Kauffmann, Marina Vendramini, Amanda M V Batista, Helena B S Mota, Ikaro A Andrade, Stephanny B S Cárdenas, Paloma S Queiroz, Bruno A Silva, José R Correa, Tatsuya Nagata
Watermelon crinkle leaf-associated virus 1 (WCLaV-1) and WCLaV-2, both belonging to the genus Coguvirus (family Phenuiviridae), have been identified in watermelon plants in Brazil. To study tissue tropism and the potential for seed transmission of these viruses, we initially planned to produce specific antibodies. However, difficulties in isolating and propagating the virus in host plants hindered the purified virus preparations. To overcome this problem, the nucleocapsid (N) proteins of WCLaV-1 and -2 were produced using the pepper ringspot virus vector...
January 19, 2024: Journal of Virological Methods
https://read.qxmd.com/read/38237867/evaluation-of-long-term-preservation-methods-for-viral-rna-in-mosquitoes-at-room-temperature
#25
JOURNAL ARTICLE
Izumi Kai, Daisuke Kobayashi, Kentaro Itokawa, Chizu Sanjoba, Kyo Itoyama, Haruhiko Isawa
Mosquitoes are important vectors of various pathogenic viruses. Almost all viruses transmitted by mosquitoes are RNA viruses. Therefore, to detect viral genes, mosquito samples must be kept at low temperatures to prevent RNA degradation. However, prolonged transport from the field to laboratory can pose challenges for temperature control. The aim of this study was to evaluate methods for preserving viral RNA in mosquito bodies at room temperature. Virus-infected mosquito samples were immersed in ethanol, propylene glycol, and a commercially available nucleic acid preservation reagent at room temperature, and viral RNA stability was compared...
January 16, 2024: Journal of Virological Methods
https://read.qxmd.com/read/38228247/establishment-and-application-of-an-indirect-elisa-for-getah-virus-e2-antibody-detection
#26
JOURNAL ARTICLE
Dong You, Yu-Ling Wang, Liang-Peng Ge, Yuan-Cheng Zhou, Jing Sun, Li-Qiao Lang, Si-Yuan Lai, Yan-Ru Ai, Ling Zhu, Zhi-Wen Xu
Getah virus (GETV) is a mosquito-transmitted disease that affects animals, causing fever, aseptic meningitis, and abortion. Its prevalence in China poses risks to both animal health and public well-being. Currently, there is a scarcity of seroepidemiological data on GETV due to the absence of commercial antibody detection kits for pigs. The aim of this study is to develop a rapid, accurate, and sensitive ELISA, providing a reliable tool for GETV seroepidemiology and laying the foundation for future commercial assay development...
January 14, 2024: Journal of Virological Methods
https://read.qxmd.com/read/38218417/optimised-protocols-to-generate-high-titre-lentiviral-vectors-using-a-novel-transfection-agent-enabling-extended-hek293t-culture-following-transient-transfection-and-suspension-culture
#27
JOURNAL ARTICLE
Saqlain Suleman, Serena Fawaz, Terry Roberts, Stuart Ellison, Brian Bigger, Michael Themis
HIV-1 based lentiviral viruses are considered powerful and versatile gene therapy vectors to deliver therapeutic genes to patients with hereditary or acquired diseases. These vectors can efficiently transduce a variety of cell types when dividing or non-dividing to provide permanent delivery and long-term gene expression. Demand for scalable manufacturing protocols able to generate enough high titre vector for widespread use of this technology is increasing and considerable efforts to improve vector production cost-effectively, is ongoing...
January 11, 2024: Journal of Virological Methods
https://read.qxmd.com/read/38195043/a-method-for-screening-cdv-microneutralization-activity-in-microvolume-samples
#28
JOURNAL ARTICLE
Xiaoyu Deng, Jiazi Su, Bo Hu, Xue Bai
OBJECTIVE: This study aims to establish a screening method for canine distemper virus (CDV) microneutralizing activity suitable for microvolume samples. METHODS: This method is based on the Indirect immunofluorescence assay (IFA) established on Vero-slam cells. First, by comparing the sensitivities of CDV neutralizing monoclonal antibody (1C42H11) and NP protein monoclonal antibody (CDV-NP) in IFA experiments, CDV-NP was selected as the primary antibody. Then, by detecting the infection rates of multi-concentrations of CDV neutralized with water, the minimum CDV concentration with an infection rate greater than 90% was defined as the minimum stable infection concentration, which was used as the neutralizing solution for this method...
January 7, 2024: Journal of Virological Methods
https://read.qxmd.com/read/38184072/quantification-of-low-level-cytomegalovirus-and-epstein-barr-virus-dnaemia-by-digital-pcr
#29
JOURNAL ARTICLE
DeVon N Hunter-Schlichting, Rachel I Vogel, Melissa A Geller, Heather H Nelson
BACKGROUND: Digital PCR (dPCR) can quantify cell-free viral DNA (DNAemia), a biomarker of active viral infection. To accelerate epidemiologic investigation into low-level viral reactivation in chronic disease, we have evaluated the performance of dPCR to detect cytomegalovirus (CMV) and Epstein-Barr virus (EBV) DNAemia across platforms and blood matrices. METHODS: The droplet-based (BioRad) dPCR platform performance was compared to chip-based (BioMark), and assay validation followed dMIQE guidelines...
January 4, 2024: Journal of Virological Methods
https://read.qxmd.com/read/38176614/identification-of-a-potent-and-specific-retinoic-acid-inducible-gene-1-pathway-activator-as-a-hepatitis-b-virus-antiviral-through-a-novel-cell-based-reporter-assay
#30
JOURNAL ARTICLE
Liping Shi, Guangyang Guo, Jinying Zhou, Zhanling Cheng, Ren Zhu, George Kukolj, Chris Li
Chronic Hepatitis B Virus (HBV) infection remains a global burden. To identify small molecule RIG-I agonists as antivirals against HBV, we developed an HBV-pgRNA-based interferon-β (IFN-β) luciferase reporter assay with high level of assay sensitivity, specificity and robustness. Through HTS screening, lead compound (JJ#1) was identified to activate RIG-I signaling pathway by inducing TBK1 phosphorylation. Knockdown experiments demonstrated that JJ#1-induced retinoic acid-inducible gene 1 (RIG-I) signaling pathway activation was MAVS-dependent...
January 2, 2024: Journal of Virological Methods
https://read.qxmd.com/read/38029971/development-and-application-of-a-multiplex-pcr-method-for-the-simultaneous-detection-of-goose-parvovirus-waterfowl-reovirus-and-goose-astrovirus-in-muscovy-ducks
#31
JOURNAL ARTICLE
Shizhong Zhang, Hui Dong, Fengqiang Lin, Xiaoxia Cheng, Xiaoli Zhu, Dandan Jiang, Shifeng Xiao, Shaoying Chen, Shilong Chen, Shao Wang
A multiplex polymerase chain reaction (PCR) method was developed to detect and distinguish goose parvovirus (GPV), waterfowl reovirus (WRV), and goose astrovirus (GAstV). Three pairs of primers were designed based on conserved regions in the genomic sequences of these enteric viruses and were used to specifically amplify targeted fragments of 493 bp from the viral protein 3 (VP3) gene of GPV, 300 bp from the sigma A-encoding gene of WRV, and 156 bp from the capsid protein-encoding gene of GAstV...
February 2024: Journal of Virological Methods
https://read.qxmd.com/read/38154579/development-of-a-double-antibody-sandwich-enzyme-linked-immunosorbent-assay-for-rapid-detection-of-vzv
#32
JOURNAL ARTICLE
Aiping Wang, Na Liu, Jianguo Zhao, Yan Niu, Yumei Chen, Jingming Zhou, Enping Liu, Gaiping Zhang
BACKGROUND: Varicella zoster virus (VZV) is the pathogen of varicella and herpes zoster, it is necessary to develop a rapid, sensitive and specific detection method for the prevention and control of related diseases. METHODS: We inserted the gB protein extracellular region gene (gB-ex, 1-2208bp) of VZV into lentivirus vector, and then obtained the recombinant gB protein through mammalian expression system. BALB/c mice were immunized multiple times with purified gB protein as immunogen...
December 26, 2023: Journal of Virological Methods
https://read.qxmd.com/read/38142820/a-novel-vp1-based-enzyme-linked-immunosorbent-assay-revealed-widespread-enterovirus-g-infections-in-guangxi-china
#33
JOURNAL ARTICLE
Dalin Hong, Jinni Bian, Lingyou Zeng, Shiting Huang, Yifeng Qin, Ying Chen, Zuzhang Wei, Weijian Huang, Kang Ouyang
Enterovirus G (EV-G) has recently been shown to affect weight gain and cause neurological symptoms in piglets. However, the serological investigation of EV-G is limited. In this study, we developed a novel serological detection method based on the structural protein, VP1 of EV-G. The intra-assay and inter-assay coefficient variations were 3.2~8.9% and 2.6~8.0%, respectively. There was no cross-reaction of the VP1-based enzyme-linked immunosorbent assay (ELISA) with antisera against the other known porcine viruses...
December 22, 2023: Journal of Virological Methods
https://read.qxmd.com/read/38128833/m-pima%C3%A2-hiv1-2-vl-a-suitable-tool-for-hiv-1-and-hiv-2-viral-load-quantification-in-west-africa
#34
JOURNAL ARTICLE
Halimatou Diop-Ndiaye, Pauline Yacine Sène, Khadidiatou Coulibaly, Marième Diallo, Sada Diallo, Karim Diop, Aissatou Sow-Ndoye, Mengue Fall, Anna Julienne Selbe Ndiaye, Evans Mathebula, Adjratou Aissatou Ba, Charlotte Lejeune, Ndeye Marie Pascaline Manga, Makhtar Camara, Cheikh Tidiane Ndour, Coumba Toure Kane
Point-of-Care for HIV viral RNA quantification seems to be a complementary strategy to the existing conventional systems. This study evaluated the performance of the m-PIMA™ HIV1/2 Viral Load for the quantification of both HIV-1 and HIV-2 RNA viral load. A total of 555 HIV-1 and 90 HIV-2 samples previously tested by Abbott RealTime HIV-1 (Abbott, Chicago, USA) and Generic HIV-2® Charge virale (Biocentric, France) were tested using the m-PIMA™ HIV1/2 Viral Load at the HIV National Reference lab in Senegal...
December 19, 2023: Journal of Virological Methods
https://read.qxmd.com/read/38103738/comparison-of-rt-lamp-and-rt-qpcr-assays-for-detecting-sars-cov-2-in-the-extracted-rna-and-direct-swab-samples
#35
JOURNAL ARTICLE
Ramin Pourakbari, Mohammad Gholami, Ali Shakerimoghaddam, Farhad Motavalli Khiavi, Mojgan Mohammadimehr, Mehdi Shakouri Khomartash
Rapid detection of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) in infected patients is critical for infection control. Loop-mediated isothermal amplification (LAMP) has been demonstrated to be a rapid, simple, reliable, cost-effective and sensitive method to detect SARS-CoV-2 in a variety of samples in considerably less time than Real-Time PCR. In this study, we developed and optimized a rapid detection method for SARS-CoV-2 based on RT-LAMP method utilizing a specific primer set targeting the ORF1a gene and then examined its sensitivity and efficiency using a serially diluted viral RNA sample with a known concentration...
December 15, 2023: Journal of Virological Methods
https://read.qxmd.com/read/38086433/development-of-an-accurate-and-rapid-method-for-whole-genome-characterization-of-canine-parvovirus
#36
JOURNAL ARTICLE
Emma Condon, Sofía Grecco, Ana Marandino, Jaime Aldaz, Javier Enciso, Luis Alfaro, Danilo Bucafusco, Ruben Pérez, Yanina Panzera
Canine parvovirus is a highly contagious pathogen affecting domestic dogs and other carnivores globally. Monitoring CPV through continuous genomic surveillance is crucial for mapping variability and developing effective control measures. Here, we developed a method using multiplex-PCR-next-generation sequencing to obtain full-length CPV genomes directly from clinical samples. This approach utilizes tiling and tailed amplicons to amplify overlapping fragments of roughly 250 base pairs. This enables the creation of Illumina libraries by conducting two PCR reaction runs...
December 10, 2023: Journal of Virological Methods
https://read.qxmd.com/read/38061674/assessing-the-efficacy-of-different-bead-based-assays-in-capturing-hepatitis-e-virus
#37
JOURNAL ARTICLE
Jeremy Tan, Jennifer Harlow, Jonathon Cecillon, Neda Nasheri
Hepatitis E virus (HEV) generally causes acute liver infection in humans and its transmission could be waterborne, foodborne, bloodborne, or zoonotic. To date, there is no standard method for the detection of HEV from food and environmental samples. Herein, we explored the possibility of using magnetic beads for the capture and detection of HEV. For this purpose, we employed Dynabeads M-270 Epoxy magnetic beads, coated with different monoclonal antibodies (mAbs) against HEV capsid protein, and the Nanotrap Microbiome A Particle magnetic beads, which are coated with chemical affinity baits, to capture HEV-3 particles in suspension...
December 5, 2023: Journal of Virological Methods
https://read.qxmd.com/read/38061673/comparison-of-quantitative-pcr-and-digital-pcr-assays-for-quantitative-detection-of-infectious-bronchitis-virus-ibv-genome
#38
JOURNAL ARTICLE
Ishara M Isham, Shahnas M Najimudeen, Susan C Cork, Ashish Gupta, Mohamed Faizal Abdul-Careem
The quantitative polymerase chain reaction (qPCR) technique is an extensively used molecular tool for the detection and quantification of viral genome load. However, since qPCR is a relative quantification method that relies on an external calibration curve it has a lower assay precision and sensitivity. The digital PCR (dPCR) technique is a good alternative to the qPCR assay as it offers highly precise and direct quantification of viral genome load in samples. In this study, performance characteristics such as the quantification range, sensitivity, precision, and specificity of the dPCR technique was compared to qPCR technique for the detection and quantification of IBV genome loads in serial dilutions of IBV positive plasmid DNA, and IBV infected chicken tissue and swab samples...
December 5, 2023: Journal of Virological Methods
https://read.qxmd.com/read/38029970/detection-of-hiv-1-matrix-protein-p17-in-sera-of-viremic-or-aviremic-patients
#39
JOURNAL ARTICLE
Alberto Zani, Serena Messali, Matteo Uggeri, Carlo Bonfanti, Arnaldo Caruso, Francesca Caccuri
People living with human immunodeficiency virus type 1 (HIV-1), even if successfully treated with a combined antiretroviral therapy, display a persistent inflammation and chronic immune activation, and an increasing risk of developing cardiovascular and thrombotic events, cancers, and neurologic disorders. Accumulating evidence reveals that biologically active HIV-1 proteins may play a role in the development of these HIV-1-associated conditions. The HIV-1 matrix protein p17 (p17) is released and accumulates in different organs and tissue where it may exert multiple biological activities on different target cells...
November 27, 2023: Journal of Virological Methods
https://read.qxmd.com/read/38013021/screening-and-identification-of-b-cell-epitope-within-the-major-capsid-protein-l1-of-hpv-52-using-monoclonal-antibodies
#40
JOURNAL ARTICLE
Yumei Chen, Shan Zhang, Gaiping Zhang, Jingming Zhou, Hongliang Liu, Chao Liang, Enping Liu, Xifang Zhu, Aiping Wang
The L1 protein of Human papillomavirus (HPV), the main capsid protein, induces the formation of neutralizing antibodies. In this study, HPV52 L1 protein was induced to be expressed. Monoclonal antibody (mAb) 6A7 against L1 protein were screened by cell fusion techniques. Western Blot and immunofluorescence assay (IFA) demonstrated the specificity of the mAb. The L1 protein was truncated for prokaryotic expression (N1~N7) and Dot-ELISA showed that 6A7 recognized N3 (aa 200~350). The immunodominant regions were truncated again for expression, with 6A7 recognizing N6 (aa 251~305)...
November 25, 2023: Journal of Virological Methods
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