Abstract：
:Epidemics of acute hemorrhagic conjunctivitis are always explosive and extensive, and have been recognized as a serious international public health problem. Enterovirus 70 and coxsackievirus A24 variant have been identified as the major etiological agents in acute hemorrhagic conjunctivitis outbreaks worldwide. A novel multiplex real-time RT-PCR assay was developed for simultaneous detection, identification and quantitation of enterovirus 70 and coxsackievirus A24 variant. The specificity, sensitivity and reproducibility of the method were analyzed and 125 clinical samples were tested using this method. No cross-reactivity with other enteroviruses strains was detected. The detection limits achieved were 10 copies/tube of enterovirus 70 and 100 copies/tube of coxsackievirus A24 variant respectively, and the addition of the internal control does not compromise the sensitivity or specificity. One hundred and twenty five clinical samples were tested and the results were consistent with the results obtained by using virus isolation followed by neutralization and sequencing of VP1 region. Due to its high specificity, sensitivity and elimination of false negative results by the internal control, this assay is suitable for both research applications and rapid clinical diagnosis of enterovirus 70 and coxsackievirus A24 variant.
journal_name
J Virol Methodsjournal_title
Journal of virological methodsauthors
Xiao XL,Wu H,Li YJ,Li HF,He YQ,Chen G,Zhang JW,Yang H,Li XF,Yang XQ,Yu YGdoi
10.1016/j.jviromet.2009.02.022subject
Has Abstractpub_date
2009-07-01 00:00:00pages
23-8issue
1eissn
0166-0934issn
1879-0984pii
S0166-0934(09)00095-0journal_volume
159pub_type
杂志文章abstract：:Performances of serological parallel and serial testing algorithms were analyzed using a combination of three ELISA and three rapid tests for the confirmation of HIV infection. Each was assessed individually for their sensitivity and specificity on a blinded panel of 769 retrospective sera of known HIV status. Western...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/j.jviromet.2011.05.021
更新日期：2011-09-01 00:00:00
abstract：:Hexon is the major structural protein of adenovirus, and has significance in studies of virus structure and function, vaccine development, and immunodiagnosis. We describe a simple, single-step, anion-exchange high performance liquid chromatography (HPLC) method for the high yield purification of hexon. Purity of the ...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/0166-0934(87)90131-5
更新日期：1987-09-01 00:00:00
abstract：:Ten plasmids containing viral cDNA inserts of portions of the dengue virus type 2 (DEN-2) or Kunjin virus (KUN) genomes were biotinylated using photobiotin acetate and used as probes for the detection of flavivirus RNA in infected Vero cells. The viral cDNA inserts ranged in length from 0.19 to 2.7 kilobase pairs, and...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/0166-0934(87)90055-3
更新日期：1987-02-01 00:00:00
abstract：:Integration of reverse transcribed viral DNA of HIV into host chromosomes is mediated by the viral enzyme, integrase. This enzymatic activity can be monitored in vitro by integration of a small labeled DNA (donor) into a second unlabeled DNA (target). The methodology usually involves isotope labeling and gel electroph...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/0166-0934(96)02033-2
更新日期：1996-05-01 00:00:00
abstract：:Simultaneous detection of two antigens by immunostaining usually requires primary antibodies from two different species or a hapten modification of one of the antibodies if they are from the same species. A novel double staining method is described for immunodetection of two independent antigens using two mouse monocl...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/s0166-0934(98)00048-2
更新日期：1998-07-01 00:00:00
abstract：:Marek's disease virus (MDV) is a highly oncogenic alphaherpesvirus that induces rapid-onset T-cell lymphoma in poultry. The complete genome of the avirulent vaccine strain MDV-814 was cloned as an infectious bacterial artificial chromosome (BAC) using an 8.8-kb fragment containing the self-designed selective marker gu...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/j.jviromet.2008.10.021
更新日期：2009-03-01 00:00:00
abstract：:Bovine viral diarrhea virus (BVDV) can contaminate biological products produced in bovine or porcine cells or manufactured using bovine sera. A rapid, specific, sensitive, and practical method of detecting BVDV in bio-products is needed. The purpose of this study was to compare three assays with respect to their abili...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/j.jviromet.2014.05.020
更新日期：2014-10-01 00:00:00
abstract：:Double-stranded RNA (dsRNA) molecules of viruses are found in nature at a very high frequency. Their detection in plants and fungi has been carried out with difficulty due to the complicated dsRNA extraction techniques used commonly which includes phenol-chloroform extractions. In this study, an extraction method for ...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/j.jviromet.2008.06.001
更新日期：2008-09-01 00:00:00
abstract：:A reverse transcription loop-mediated isothermal amplification (RT-LAMP) assay was developed for the rapid detection of equine coronavirus (ECoV). This assay was conducted at 60 °C for 40 min. Specificity of the RT-LAMP assay was confirmed using several equine intestinal and respiratory pathogens in addition to ECoV. ...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/j.jviromet.2015.02.001
更新日期：2015-04-01 00:00:00
abstract：:Paramyxoviruses, including measles virus (MV), human metapneumovirus (HMPV), human respiratory syncytial virus (HRSV) and canine distemper virus (CDV), are transmitted via the respiratory route. Despite their close phylogenetic relationship, the pathogenesis of these viruses is very different. To study viral tropism a...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/j.jviromet.2013.06.016
更新日期：2013-10-01 00:00:00
abstract：:Rotavirus (RoV) and Norovirus (NoV) are the main causes of viral gastroenteritis. Currently, there is no validated multiplex real-time PCR that can detect and quantify RoV and NoV simultaneously. The aim of the study was to develop, validate, and internally control a multiplex one-step RT real-time PCR to detect and q...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/j.jviromet.2012.09.021
更新日期：2013-01-01 00:00:00
abstract：:Rice stripe virus (RSV), Rice black-streaked dwarf virus (RBSDV), and Rice dwarf virus (RDV) are major rice-infecting viruses in Korea that can cause serious crop losses. A one-step multiplex reverse transcription-polymerase chain reaction (mRT-PCR) was developed for the simultaneous detection of these rice viruses. T...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/j.jviromet.2013.07.008
更新日期：2013-11-01 00:00:00
abstract：:A method for rapid and reliable detection of Tomato spotted wilt virus (TSWV) (Tospovirus, Bunyaviridae) in its vector Frankliniella occidentalis (Thysanoptera Thripidae) would be a useful tool for studying the epidemiology of this virus. A RT-PCR method developed for this purpose is reported. The method was tested on...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/s0166-0934(03)00048-x
更新日期：2003-04-01 00:00:00
abstract：:Droplet Digital PCR (ddPCR) represents a new and alternative platform to conventional quantitative-PCR (qPCR) for the quantitation of DNA templates. However, the proposed improvement in sensitivity and reproducibility offered by ddPCR is not yet fully proven, partly because the delineation between positive and negativ...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/j.jviromet.2014.02.020
更新日期：2014-06-01 00:00:00
abstract：:Diagnostic tests are usually evaluated in terms of simple qualitative measures of sensitivity and specificity. When comparing different quantitative assays such as ELISAs, it is often more useful to deal with actual values (sample optical density/cut-off optical density ratio (OD ratio] rather than the qualitative rel...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/0166-0934(88)90087-0
更新日期：1988-10-01 00:00:00
abstract：:In view of the genetic diversity of the human immunodeficiency virus Type 1, we assessed the sensitivity and quantification efficiency of the HIV-1 RNA NASBA amplification system with respect to different HIV-1 subtypes and recombinants. Twenty cell culture supernatants representing 17 HIV-1 group M and 3 group O stra...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/s0166-0934(97)00072-4
更新日期：1997-07-01 00:00:00
abstract：:PRRSV is a positive-sense RNA virus with a high degree of genetic variability among isolates. For diagnostic sensitivity and vaccine design it is essential to monitor PRRSV genetic diversity. However, to date only a few full genome sequences of PRRSV isolates have been made publicly available. In the present study, fa...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/j.jviromet.2013.07.019
更新日期：2013-11-01 00:00:00
abstract：:A solid phase reverse passive hemadsorption test (SP-RPHAd) for hepatitis B surface antigen detection is described. It was compared with a commercial reverse passive hemagglutination assay (Hepatest, Wellcome, U.K.). SP-RPHAd is four-fold less expensive than Hepatest and undiluted sera can be used instead of eight-fol...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/0166-0934(87)90115-7
更新日期：1987-11-01 00:00:00
abstract：:A rapid and accurate detection and identification system was developed for flaviviruses that makes use of reverse transcription-polymerase chain reaction (RT-PCR). A primer pair (YF-1 and YF-3), which corresponds to the highly conserved sequence at 3' noncoding region among flaviviruses, was useful for identification ...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/0166-0934(93)90020-r
更新日期：1993-03-01 00:00:00
abstract：:Four groups of cattle were tested for antibodies against foot-and-mouth disease (FMD) virus type O(1) over three 70 day vaccination cycles using the liquid-phase-blocking-ELISA (LPBE). First lactation cows showed the lowest titres and group protection levels (GPLs) against FMD virus strains with 'r' values < or =0.5 w...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/s0166-0934(01)00342-1
更新日期：2001-09-01 00:00:00
abstract：:A polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) assay has been developed to detect and differentiate between canine parvovirus (CPV) and mink enteritis virus (MEV). Eight CPV and three MEV epidemic strains isolated from 28 pathological samples from dogs and minks suspected of being infe...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/j.jviromet.2014.09.014
更新日期：2014-12-15 00:00:00
abstract：:Quantitative real-time PCR is being widely used in the identification of plasma donations that contain high levels of parvovirus B19, to ensure their exclusion from start pools used in the manufacture of plasma derived medicinal products. In this study, the primers and probe of one such published assay, are examined f...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/j.jviromet.2006.09.002
更新日期：2007-01-01 00:00:00
abstract：:A recombinant murine cytomegalovirus (mCMV) that expresses enhanced green fluorescent protein (EGFP) under control of the native immediate-early 1/3 promoter was constructed to detect directly sites of viral activity in latent and reactivated infections. The recombinant virus had acute and latent infection characteris...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/s0166-0934(00)00202-0
更新日期：2000-09-01 00:00:00
abstract：:Goatpox disease and cystic hydatidosis may be simultaneously endemic in a given area. Their pathogens are goatpox virus (GPV) and Echinococcus granulosus (E. granulosus), respectively. Both E. granulosus EG95 subunit vaccine and live-attenuated GPV AV41 vaccine have been widely used for prevention of both diseases in ...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/j.jviromet.2018.08.002
更新日期：2018-11-01 00:00:00
abstract：:A novel isothermal multiplex AmpliDet RNA system is described for the simultaneous amplification and detection of Potato leafroll virus (PLRV) and Potato virus Y (PVY) in seed potatoes. The risk of contamination by carry-over during diagnostic screening is eliminated by performing the reaction in a single closed tube....
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/s0166-0934(01)00258-0
更新日期：2001-04-01 00:00:00
abstract：:This study describes the development and evaluation of a cost effective test rationale for the detection of anti-HCV in dried blood spots. Samples were screened using an 'in house' IgG ELISA that incorporated the recombinant proteins c22-3, c200 and NS5. Confirmation of specific antibody to HCV was by a modification o...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/s0166-0934(97)00127-4
更新日期：1997-11-01 00:00:00
abstract：:Studies were performed to identify a pair of primers, specific for the tick-borne encephalitis (TBE) virus complex of the Flaviviridae, with which to develop a rapid and specific identification system based on reverse transcription and the polymerase chain reaction (RT-PCR). The specificity of a putative primer pair w...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/0166-0934(93)90144-g
更新日期：1993-11-01 00:00:00
abstract：:Plaque assays under Sephadex or agarose overlays are described for rat coronaviruses (RCVs) grown in L2 mouse fibroblasts. A plaque assay using Sephadex was simple; however, viable plaques could not be collected for propagation, and fixation was necessary before evaluation. Plaque formation under agarose was optimized...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/0166-0934(93)90089-a
更新日期：1993-06-01 00:00:00
abstract：:Canine distemper virus (CDV) is an important pathogen of many carnivores. We are developing a field-based model of morbillivirus virulence and pathogenesis through a study of distemper in naturally infected free-ranging raccoons. The isolation of CDV from raccoon tissues is essential for this work. CDV has often been ...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/j.jviromet.2004.02.004
更新日期：2004-06-15 00:00:00
abstract：:Early detection of resistant mutations of hepatitis B virus (HBV) is important for patients on nucleos(t)ide analog therapy. An assay based on the PCR-Invader technology was developed to detect resistant mutations with high sensitivity. The assay specifically detects mutations at codons 180, 181, 184, 202, 204, and 25...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/j.jviromet.2010.10.001
更新日期：2011-01-01 00:00:00
