Abstract：
:The SPF10 PCR targets a conserved 65bp region of the HPV L1 gene for broad-spectrum amplification. The LiPA assay allows subsequent genotyping of the HPV amplicons. This study aims to develop a SPF10 real-time PCR to achieve simultaneous amplification and detection of the HPV target. That way, LiPA analysis of the HPV-negative samples can be avoided, reducing workload and cost. The real-time PCR shows an analytical sensitivity of 29.7 copies for HPV 6, 16, 18 and 31 and an HPV-specific melting peak. Thirty-one HPV DNA plasmids were genotyped correctly using the SPF10 real-time PCR in combination with the LiPA. Here, the LiPA assay was performed at an increased hybridisation temperature (49.5°C) in combination with a reduced amplicon volume (1μl) to avoid cross-reactivity. In conclusion, the SPF10 real-time PCR proves to be very sensitive and generates amplicons, which are compatible with the LiPA.
journal_name
J Virol Methodsjournal_title
Journal of virological methodsauthors
Micalessi MI,Boulet GA,Vorsters A,De Wit K,Jannes G,Mijs W,Ieven M,Van Damme P,Bogers JJdoi
10.1016/j.jviromet.2012.09.013subject
Has Abstractpub_date
2013-01-01 00:00:00pages
166-71issue
1eissn
0166-0934issn
1879-0984pii
S0166-0934(12)00335-7journal_volume
187pub_type
杂志文章abstract：:The Cucumber mosaic virus (CMV) is an isodiametric plant virus with an extremely wide host range, present worldwide. CMV chimeric particles (R9-CMV), engineered to express a 27-aa synthetic peptide derived from Hepatitis C virus (HCV), were demonstrated to be stable under simulated gastric and intestinal conditions. T...
journal_title：Journal of virological methods
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abstract：:The Pr55gag human immunodeficiency virus type 1 (HIV-1) precursor protein that is capable of auto-assembling was used as a carrier for a consensus sequence of the principal neutralization domain (PND) of the HIV-1 envelope. For this purpose, a modified HIV-1 gag gene with deletion of the sequence encoding a previously...
journal_title：Journal of virological methods
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doi：10.1016/0166-0934(94)00100-u
更新日期：1995-02-01 00:00:00
abstract：:A recombinant retroviral system was used for the analysis of early HIV breakthrough infection in the presence of antiviral drugs. The use of replication-defective HIV allowed a quantitative analysis of a single cycle of infection. This report characterizes this recombinant HIV system and demonstrates it's validity in ...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/s0166-0934(97)00212-7
更新日期：1998-04-01 00:00:00
abstract：:The occurrence of virus belonging to the putative genus Influenzavirus D, has been demonstrated all-around the world arousing interest within the scientific community. Most of the published virological surveys are based on the first described Real-Time PCR method, designed on the PB1 gene of the first isolate. The nec...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/j.jviromet.2017.01.019
更新日期：2017-05-01 00:00:00
abstract：:To develop reverse genetics system of RNA viruses, cloning of full-length viral genome is required which is often challenging due to many steps involved. In this study, we report cloning of full-length cDNA from an Indian field isolate (CSFV/IVRI/VB-131) of classical swine fever virus (CSFV) using in vitro overlap ext...
journal_title：Journal of virological methods
pub_type： 杂志文章
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abstract：:Different viruses belonging to the genus Vesivirus infect a broad range of animals, and cause gastroenteritis, vesicular skin lesions, hemorrhagic disease, respiratory diseases and other conditions. A recent report on Vesivirus viremia, as detected by PCR, in samples from patients with hepatitis of unknown etiology in...
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doi：10.1016/j.jviromet.2008.12.008
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journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/j.jviromet.2009.05.021
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journal_title：Journal of virological methods
pub_type： 临床试验,杂志文章
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更新日期：2006-09-01 00:00:00
abstract：:Rapid characterization of herpesvirus isolates exemplified by equine herpesviruses is described. Total DNAs were isolated from virus infected small scale cell cultures. The DNA fragments obtained after restriction enzyme digestion were separated on agarose gels, transferred and immobilized on filter membranes. A radio...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/0166-0934(86)90030-3
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abstract：:Antigenaemia due to human immunodeficiency virus (HIV) is thought to be significant either before the appearance of a specific antibody response, or after its decline during terminal stages. In order to increase the rate of detection of HIV antigen carriers, regardless of the stage or despite the presence of specific ...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/0166-0934(88)90115-2
更新日期：1988-12-01 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：:The major protective antigens of retroviruses are considered to be their glycosylated envelope proteins. However, the methods commonly employed to enrich and purify virus from culture media such as pelleting and density-gradient centrifugation result in a low recovery of the viral external glycoproteins. This is an ob...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/0166-0934(89)90011-6
更新日期：1989-04-01 00:00:00
abstract：:A TaqMan real-time RT-PCR was developed to detect and quantify RNA-targets from the non-circulative, non-persistently transmitted Plum pox virus (PPV) in individual fresh or aphids captured previously and squashed on paper. Reliable quantitation ranged from 40 up to 4 x 10(8) copies of control transcripts. This techni...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/j.jviromet.2005.05.011
更新日期：2005-09-01 00:00:00
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journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/s0166-0934(03)00079-x
更新日期：2003-05-01 00:00:00
abstract：:Participation in external quality assessment programs is critical to ensure quality clinical laboratory testing. Commercially available proficiency test panels for HIV-1 virus load testing that are used commonly in external quality assessment programs remain a financial obstacle to resource-limited countries. Maintain...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/j.jviromet.2012.11.036
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abstract：:HIV-1 RNA viral load has become the major biological marker for disease prognosis and outcome of antiretroviral therapy in the treatment of HIV-infected individuals. The aim of this study was to compare the performance of the new CE marked NucliSens EasyQ HIV-1 assay with NucliSens HIV-1 QT assay (reference method). N...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/j.jviromet.2005.04.017
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journal_title：Journal of virological methods
pub_type： 杂志文章
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journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/0166-0934(87)90011-5
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abstract：:Human cytomegalovirus (HCMV) is the most frequent cause of congenital infection. Diagnosis of this infection is important because 5-17% of asymptomatic infected babies will develop late sequelae and should be followed closely. Most of these children will remain undetected, since screening of all newborns by viral cult...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/j.jviromet.2005.02.018
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journal_title：Journal of virological methods
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doi：10.1016/j.jviromet.2009.11.033
更新日期：2010-05-01 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
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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 RT-nested PCR that amplifies part of the conserved nucleoprotein gene of avian Paramyxovirus type 1 is described. The technique allowed the detection of pigeon Paramyxovirus type 1 (pPMV-1) virus directly from a wide range of infected chicken and pigeon organs, and should be able to detect typical Newcastle disease ...
journal_title：Journal of virological methods
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abstract：:The whole nucleotide sequence of Oropouche virus medium (M) RNA, Orthobunyavirus genus, Bunyaviridae family, was obtained using a new genomic amplification method. This method is based on the use of a single and specific primer of high melting temperature in a linear amplification (LA), followed by a single primer pol...
journal_title：Journal of virological methods
pub_type： 杂志文章
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更新日期：2004-01-01 00:00:00
abstract：:HIV-1 infection cannot be cured due to the presence of the latent reservoir (LR). Novel cure or treatment strategies, such as "shock and kill" or therapeutic vaccination, aim to reduce or eradicate the LR. Cure strategies utilise robust DNA quantification assays to measure the change in the LR in low copy scenarios. N...
journal_title：Journal of virological methods
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更新日期：2019-01-01 00:00:00
abstract：:Marine caliciviruses form a distinct lineage within the genus Vesivirus (family Caliciviridae). This group includes vesicular exanthema of swine virus (VESV) and San Miguel sea lion virus (SMSV) and other related viruses which have been proposed to be marine in origin isolated from a variety of terrestrial and marine ...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/j.jviromet.2006.11.010
更新日期：2007-03-01 00:00:00
abstract：:The polymerase chain reaction (PCR) is one of the most efficient techniques for measuring the viral load of HIV-infected samples. Determination of the specificity of PCR products is usually based on Southern blotting and hybridization of the amplified DNA to radioactive oligonucleotide probes specific for sequences co...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/0166-0934(94)90028-0
更新日期：1994-05-01 00:00:00
abstract：:The rapid development of new molecular biology methods has improved infectious disease diagnosis, which is increasingly important to clinical management and public health. A wide variety of new methods which are more specific, sensitive and robust, such as combination of PCR and microarray technology, has gradually re...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/j.jviromet.2007.05.018
更新日期：2007-11-01 00:00:00
abstract：:In this inhibition immunoassay undiluted serum reacts in solution with crude cellular CMV antigen in wells of microtestplates coated with hyperimmune CMV-reactive monkey IgG. CMV antibodies in the serum under test block (completely or partial) the fixation of antigen to the capture layer. Unblocked antigenic activity ...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/0166-0934(87)90004-8
更新日期：1987-06-01 00:00:00
