Abstract:
BACKGROUND:Few studies have used quantitative polymerase chain reaction (qPCR) as an approach to measure virus neutralization assay endpoints. Its lack of use may not be surprising considering that sample nucleic acid extraction and purification can be expensive, labor-intensive, and rate-limiting. METHODS:Virus/antibody mixtures were incubated for one hour at 37°C and then transferred to Vero cell monolayers in a 96-well plate format. At 24 (or 48) hours post-infection, we used a commercially available reagent to prepare cell lysates amenable to direct analysis by one-step SYBR Green quantitative reverse transcription PCR using primers specific for the RSV-N gene, thereby obviating the need for cumbersome RNA extraction and purification. The neutralization titer was defined as the reciprocal of the highest dilution needed to inhibit the PCR signal by 90% when compared with the mean value observed in virus control wells in the absence of neutralizing antibodies. RESULTS:We have developed a qPCR-based neutralization assay for human respiratory syncytial virus. Due to the sensitivity of qPCR in detecting virus replication, endpoints may be assessed as early as 24 hours post-infection. In addition, the dynamic range of qPCR provides a basis for the assay to be relatively robust to perturbations in input virus dose (i.e., the assay is in compliance with the Percentage Law). CONCLUSIONS:This qPCR-based neutralization assay is suitable for automated high-throughput applications. In addition, our experimental approach may be generalizable for the rapid development of neutralization assays for other virus families.
journal_name
Virol Jjournal_title
Virology journalauthors
Varada JC,Teferedegne B,Crim RL,Mdluli T,Audet S,Peden K,Beeler J,Murata Hdoi
10.1186/1743-422X-10-195subject
Has Abstractpub_date
2013-06-15 00:00:00pages
195issn
1743-422Xpii
1743-422X-10-195journal_volume
10pub_type
杂志文章相关文献
Virology Journal文献大全abstract:BACKGROUND:The rapid and accurate identification of the H5 and H7 subtypes of avian influenza (AI) virus is an important step for the control and eradication of highly pathogenic AI outbreaks and for the surveillance of AI viruses that have the potential to undergo changes in pathogenicity in poultry and wild birds. Cu...
journal_title:Virology journal
pub_type: 杂志文章
doi:10.1186/1743-422X-10-85
更新日期:2013-03-14 00:00:00
abstract:BACKGROUND:Xenotransplantation using pig cells, tissues and organs may help to overcome the shortage of human tissues and organs for the treatment of tissue and organ failure. Progress in the prevention of immunological rejection using genetically modified pigs and new, more effective, immunosuppression regimens will a...
journal_title:Virology journal
pub_type: 杂志文章,评审
doi:10.1186/s12985-018-1088-2
更新日期:2018-11-08 00:00:00
abstract:AIM:To investigate the enhancement of humoral immunity when CpG ODN (cytidine phosphate guanosine oligodeoxynucleotides) and aluminium adjuvants are complexed with the HCV (Hepatitis C virus) recombinant immunogen in mice. METHODS:After immunizing Balb/c mice with the recombination HCV antigen adjuvanted with pUCpGs10...
journal_title:Virology journal
pub_type: 杂志文章
doi:10.1186/1743-422X-8-507
更新日期:2011-11-04 00:00:00
abstract:BACKGROUND:Viruses included in the family Iridoviridae are large, icosahedral, dsDNA viruses that are subdivided into 5 genera. Frog virus 3 (FV3) is the type species of the genus Ranavirus and the best studied iridovirus at the molecular level. Typically, antibodies directed against a virus act to neutralize the virus...
journal_title:Virology journal
pub_type: 杂志文章
doi:10.1186/1743-422X-7-41
更新日期:2010-02-18 00:00:00
abstract:BACKGROUND:Aleutian mink disease virus (AMDV) is the cause of a chronic immune complex disease, Aleutian disease (AD), which is common in mink-producing countries. In 2005, implementation of an AMDV eradication programme in Finland created a need for an automated high-throughput assay. The aim of this study was to vali...
journal_title:Virology journal
pub_type: 杂志文章
doi:10.1186/1743-422X-11-141
更新日期:2014-08-08 00:00:00
abstract:BACKGROUND:Pear ring rot, caused by Botryosphaeria species, is responsible for substantial economic losses by causing severe recession of pear tree growth in China. Mycovirus-mediated hypovirulence in plant pathogenic fungi is a crucial biological method to control fungal diseases. METHODS:We conducted a large-scale a...
journal_title:Virology journal
pub_type: 杂志文章
doi:10.1186/s12985-018-1033-4
更新日期:2018-08-13 00:00:00
abstract:BACKGROUND:Human herpesviruses have been hypothesized as environmental triggers in the development of autoimmune thyroid diseases (AITD), and in particular active human herpesvirus 6A (HHV-6A) infection was detected in thyrocytes of Hashimoto's thyroiditis (HT) patients, who also show specific anti-viral immune respons...
journal_title:Virology journal
pub_type: 杂志文章
doi:10.1186/s12985-016-0672-6
更新日期:2017-01-11 00:00:00
abstract:BACKGROUND:Acute gastroenteritis caused by bacteria, virus and parasite is an important cause of childhood morbidity and mortality in developing countries. Rotavirus and norovirus have been recognized as the most common pathogens causing acute gastroenteritis among children. However, there is still no valuable data abo...
journal_title:Virology journal
pub_type: 杂志文章
doi:10.1186/1743-422X-10-302
更新日期:2013-10-08 00:00:00
abstract::The NanoSight LM10 with Nanoparticle tracking analysis (NTA) software was evaluated for the quantification of latex particles, adenovirus 5, and influenza virus. The inter-day variability was determined by measuring the same sample over several consecutive days and the method's accuracy was demonstrated by using known...
journal_title:Virology journal
pub_type: 杂志文章
doi:10.1186/1743-422X-9-265
更新日期:2012-11-12 00:00:00
abstract:BACKGROUND:The membrane topology and molecular mechanisms for endoplasmic reticulum (ER) localization of classical swine fever virus (CSFV) non-structural 2 (NS2) protien is unclear. We attempted to elucidate the subcellular localization, and the molecular mechanisms responsible for the localization of this protein in ...
journal_title:Virology journal
pub_type: 杂志文章
doi:10.1186/1743-422X-8-236
更新日期:2011-05-18 00:00:00
abstract:BACKGROUND:Human cytomegalovirus (HCMV) is the most common cause of intrauterine infections worldwide. The toll-like receptors (TLRs) have been reported as important factors in immune response against HCMV. Particularly, TLR2, TLR4 and TLR9 have been shown to be involved in antiviral immunity. Evaluation of the role of...
journal_title:Virology journal
pub_type: 杂志文章
doi:10.1186/s12985-017-0730-8
更新日期:2017-03-24 00:00:00
abstract:BACKGROUND:Duck circovirus (DuCV) infection in farmed ducks is associated with growth problems or retardation syndromes. Rapid identification of DuCV infected ducks is essential to control DuCV effectively. Therefore, this study aims to develop of an assay for DuCV to be highly specific, sensitive, and simple without a...
journal_title:Virology journal
pub_type: 杂志文章
doi:10.1186/1743-422X-11-76
更新日期:2014-04-29 00:00:00
abstract::Virus taxonomy at present is best characterized as a categorization of convenience, without a firm basis in the principles of evolutionary biology. Specifically, virus species definitions appear to depend more on tradition and popular opinion among virologists than on firm, quantitative biological evidence. I suggest ...
journal_title:Virology journal
pub_type: 信件
doi:10.1186/1743-422X-11-131
更新日期:2014-07-23 00:00:00
abstract:BACKGROUND:Inteins are "protein introns" that remove themselves from their host proteins through an autocatalytic protein-splicing. After their discovery, inteins have been quickly identified in all domains of life, but only once to date in the genome of a eukaryote-infecting virus. RESULTS:Here we report the identifi...
journal_title:Virology journal
pub_type: 杂志文章
doi:10.1186/1743-422X-2-8
更新日期:2005-02-11 00:00:00
abstract:BACKGROUND:Local epidemiology of Dengue is defined by the genetic diversity of the circulating Dengue virus (DENV) strains. This important information is not available for the virus strains from most parts of the Indian subcontinent. The present study focused on the genetic diversity of the serotype 3 DENV strains (DEN...
journal_title:Virology journal
pub_type: 杂志文章
doi:10.1186/1743-422X-10-37
更新日期:2013-01-29 00:00:00
abstract:BACKGROUND:Sheeppox (SPP) and goatpox (GTP) caused by sheeppox virus (SPPV) and goatpox virus (GTPV), respectively of the genus Capripoxvirus in the family Poxviridae, are severely afflicting small ruminants' production systems in Africa and Asia. In endemic areas, SPP and GTP are controlled using vaccination with live...
journal_title:Virology journal
pub_type: 杂志文章
doi:10.1186/s12985-018-0969-8
更新日期:2018-04-02 00:00:00
abstract:BACKGROUND:Resonant microwave absorption has been proposed in the literature to excite the vibrational states of microorganisms in an attempt to destroy them. But it is extremely difficult to transfer microwave excitation energy to the vibrational energy of microorganisms due to severe absorption of water in this spect...
journal_title:Virology journal
pub_type: 杂志文章
doi:10.1186/1743-422X-4-50
更新日期:2007-06-05 00:00:00
abstract:BACKGROUND:Torque Teno Virus (TTV) was the first single stranded circular DNA virus to be discovered that infects humans. Although there have been numerous reports regarding the prevalence of TTV from other countries of South Asia, there is severe lack of information regarding its prevalence in Pakistan. Thus the prese...
journal_title:Virology journal
pub_type: 杂志文章
doi:10.1186/1743-422X-9-320
更新日期:2012-12-28 00:00:00
abstract:BACKGROUND:Hepatitis C virus (HCV) encodes two transmembrane glycoproteins E1 and E2 which form a heterodimer. E1 is believed to mediate fusion while E2 has been shown to bind cellular receptors including CD81. In this study, alanine substitutions in E2 were generated within putative CD81 binding regions to define resi...
journal_title:Virology journal
pub_type: 杂志文章
doi:10.1186/1743-422X-5-46
更新日期:2008-03-20 00:00:00
abstract:BACKGROUND:Infection with H5N1 highly pathogenic avian influenza viruses (HPAIVs) of domestic poultry and wild birds has spread to more than 60 countries in Eurasia and Africa. It is concerned that HPAIVs may be perpetuated in the lakes in Siberia where migratory water birds nest in summer. To monitor whether HPAIVs ci...
journal_title:Virology journal
pub_type: 杂志文章
doi:10.1186/1743-422X-8-65
更新日期:2011-02-11 00:00:00
abstract:BACKGROUND:Porcine sapovirus was first identified in the United States in 1980, hitherto, several Asian countries have detected this virus. In 2008, the first outbreak of gastroenteritis in piglets caused by porcine sapovirus in China was reported. The complete genome of the identified SaV strain Ch-sw-sav1 was sequenc...
journal_title:Virology journal
pub_type: 杂志文章
doi:10.1186/1743-422X-6-216
更新日期:2009-12-06 00:00:00
abstract:BACKGROUND:Cell culture adaptation of very virulent infectious bursal disease virus (vvIBDV) was shown to be mainly associated with the VP2 capsid protein residues 253, 279, and 284. The single mutation A284T proved critical for cell culture tropism, but did not confer efficient virus replication, which at least requir...
journal_title:Virology journal
pub_type: 杂志文章
doi:10.1186/s12985-014-0199-7
更新日期:2014-11-25 00:00:00
abstract:BACKGROUND:In mammals, interleukin 2 (IL-2) has been shown to decrease replication or attenuate pathogenicity of numerous viral pathogens (herpes simplex virus, vaccinia virus, human respiratory syncytial virus, human immunodeficiency virus) by activating natural killer cells (NK), cytotoxic T lymphocytes and expanding...
journal_title:Virology journal
pub_type: 杂志文章
doi:10.1186/s12985-015-0353-x
更新日期:2015-08-08 00:00:00
abstract:BACKGROUND:A novel reassortant H7N9 influenza A virus has crossed the species barrier from poultry to cause human infections in China in 2013 and 2014. Rapid detection of the novel H7N9 virus is important to detect this virus in poultry and reduce the risk of an epidemic in birds or humans. FINDINGS:In this study, a m...
journal_title:Virology journal
pub_type: 杂志文章
doi:10.1186/s12985-015-0300-x
更新日期:2015-04-30 00:00:00
abstract:BACKGROUND:Human noroviruses (NoVs) are the main cause of gastroenteritis worldwide. The most commonly detected NoV strains belong to the genetically diverse GII.4 genotype, with new pandemic variants emerging periodically. Despite extensive efforts, NoV investigation has been hampered by the lack of an effective in vi...
journal_title:Virology journal
pub_type: 杂志文章
doi:10.1186/s12985-016-0538-y
更新日期:2016-05-21 00:00:00
abstract:BACKGROUND:The global spread of the highly pathogenic avian influenza H5N1 virus has stimulated interest in a better understanding of the mechanisms of H5N1 dispersal, including the potential role of migratory birds as carriers. Although wild birds have been found dead during H5N1 outbreaks, evidence suggests that othe...
journal_title:Virology journal
pub_type: 杂志文章
doi:10.1186/1743-422X-5-71
更新日期:2008-06-04 00:00:00
abstract:BACKGROUND:The human immunodeficiency virus type 1(HIV-1) is a complex retrovirus and the causative agent of acquired immunodeficiency syndrome (AIDS). The HIV-1 Vpu protein is an oligomeric integral membrane protein essential for particle release, viral load and CD4 degradation. In silico models show Vpu to form penta...
journal_title:Virology journal
pub_type: 杂志文章
doi:10.1186/1743-422X-4-81
更新日期:2007-08-29 00:00:00
abstract:UNLABELLED:In the study, a method called "hardening in liquid phase" for preparing chitosan granules with glutaraldehyde as crosslinker and Tween 80 as surfactant and paraffin liquid as dispersant was established. The chitosan granules were light yellow and insoluble in water or oil, but they swelled in acid solution a...
journal_title:Virology journal
pub_type: 杂志文章
doi:10.1186/1743-422X-7-125
更新日期:2010-06-12 00:00:00
abstract::We undertook one of the most detailed studies on the distribution of alpha2,3 sialic acid (SA)-galactose (gal) (avian type) and alpha2,6SA-gal (human type) receptors on different tissues of chickens, ducks and turkeys of varying age groups. On the tracheal epithelium, all 3 bird species expressed strong positive stain...
journal_title:Virology journal
pub_type: 杂志文章
doi:10.1186/1743-422X-7-5
更新日期:2010-01-12 00:00:00
abstract:BACKGROUND:R849 is a neurovirulent γ₁34.5 gene-deficient form of herpes simplex virus type 1 (HSV-1) and has LacZ genes at the deleted sites of the γ₁34.5 gene. HF is a spontaneously occurring, fusogenic HSV-1 strain. The purpose of this work was to generate a virus that has the syncytial character of HF, while preserv...
journal_title:Virology journal
pub_type: 杂志文章
doi:10.1186/1743-422X-8-294
更新日期:2011-06-10 00:00:00