Abstract：
:The consumption of bivalve shellfish is a common cause of foodborne outbreaks of viral origin. The evaluation of the sanitary quality of these products, however, is still based on bacterial indicators of fecal contamination (Reg. (EC) No. 2073/2005 and No.1441/2007) even if it is known that they are not reliable indicators of viral contamination. In this study a duplex Real Time PCR method for quantitative detection of hepatitis A (HAV) in shellfish was developed. Feline Calicivirus (FCV) was used as a control for assessing the effectiveness of the concentration and extraction process and the ability to eliminate PCR inhibitors present in the food matrix. The specific primers and probes for detection of HAV and FCV, chosen respectively from the 5'-UTR region and in the ORF1 region, were labeled with two different dyes and detected simultaneously. The method was applied on spiked and non-spiked shellfish from a local market. The amplification of HAV in the presence of FCV showed good linearity (R(2)=0.994) and the sensitivity limit of the reaction was at least 5 x 10(2)TCID(50)g(-1) of an hepatopancreas extract.
journal_name
J Virol Methodsjournal_title
Journal of virological methodsauthors
Di Pasquale S,Paniconi M,De Medici D,Suffredini E,Croci Ldoi
10.1016/j.jviromet.2009.09.003subject
Has Abstractpub_date
2010-01-01 00:00:00pages
96-100issue
1eissn
0166-0934issn
1879-0984pii
S0166-0934(09)00403-0journal_volume
163pub_type
杂志文章abstract：:Shrimp farming is one of the most important aquaculture activities. Expansion and intensification of shrimp farming has been accompanied with the outbreak of diseases, which threaten the development and sustainability of the industry. Viral diseases are the major challenges faced by shrimp farming industries. The prev...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/j.jviromet.2014.05.006
更新日期：2014-09-01 00:00:00
abstract：:A novel reassortant influenza A (H7N9) virus emerged recently in China. In this study, a duplex real-time reverse transcription polymerase chain reaction (rRT-PCR) assay was developed for the simultaneous detection of hemagglutinin (HA) and neuraminidase (NA) genes of H7N9 influenza viruses. The sensitivity of the ass...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/j.jviromet.2013.08.021
更新日期：2013-12-01 00:00:00
abstract：:Two enzyme-linked immunosorbent assays for detection of adenovirus antigen in faecal extracts have been established. A conjugate of rabbit anti-(human) adenovirus immunoglobulin and horseradish peroxidase (HRP) prepared by means of a hetero-bifunctional reagent, N-succinimidyl-3-(2-pyridyldithio) propionate (SPDP), wa...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/0166-0934(86)90007-8
更新日期：1986-08-01 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：:This report describes the conditions for the use of aluminum chloride (AlCl3) in growth and maintenance media for the suppression or inhibition of simian foamyviruses (SFV) in primary baboon kidney (BAK) and rabbit kidney (RK) cell cultures. When RK cells were planted in medium containing AlCl3, infected with SFV, and...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/0166-0934(82)90066-0
更新日期：1982-05-01 00:00:00
abstract：:The surface details of resin embedded viruses can be greatly enhanced by the use of uranyl acetate block staining. The virions are embedded in agar immediately after fixation and block stained with 3% uranyl acetate in methanol during dehydration. This technique allows those particles outside their host cells to show ...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/0166-0934(84)90040-5
更新日期：1984-02-01 00:00:00
abstract：:A recombinant Newcastle disease virus (NDV) expressing the green fluorescent protein (GFP) was generated by applying reverse genetics techniques. The GFP open reading frame flanked by NDV transcription start and stop sequences was inserted between the fusion (F)- and hemagglutinin-neuraminidase genes in a full-length ...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/s0166-0934(02)00247-1
更新日期：2003-03-01 00:00:00
abstract：:A near-full genome genotypic assay for HCV1b was developed, which may prove useful to investigate antiviral drug resistance, given new combination therapies for HCV1 infection. The assay consists of three partially overlapping PCRs followed by Sanger population or Illumina next-generation sequencing. Seventy-seven the...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/j.jviromet.2014.09.009
更新日期：2014-12-01 00:00:00
abstract：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 ...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/j.jviromet.2017.12.001
更新日期：2018-03-01 00:00:00
abstract：:Dengue remains a global public health threat and development of a safe and effective vaccine is a principal public health goal. The primary correlate of immunity is thought to be neutralizing antibodies. Currently, the plaque reduction neutralization test (PRNT) is the gold standard measure of dengue neutralizing anti...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/j.jviromet.2005.12.002
更新日期：2006-06-01 00:00:00
abstract：:To establish a renewable source of parvovirus B19 antigens for diagnostic tests, gene sequences for the viral capsid proteins, VP1 and VP2, were cloned into baculovirus expression vectors and the recombinant viruses used to infect Sf9 insect cells. Cell lysates examined by immunoblotting demonstrated reactive proteins...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/0166-0934(95)00046-w
更新日期：1995-09-01 00:00:00
abstract：:Beak and feather disease virus (BFDV) threatens a wide range of endangered psittacine birds worldwide. In this study, we assessed a novel PCR assay and genetic screening method using high-resolution melt (HRM) curve analysis for BFDV targeting the capsid (Cap) gene (HRM-Cap) alongside conventional PCR detection as wel...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/j.jviromet.2016.08.015
更新日期：2016-11-01 00:00:00
abstract：:Avian influenza virus (AIV) is an enveloped virus with segmented RNA that belongs to the Orthomyxoviridae. Recently, avian influenza virus isolates have not only posed a significant threat to the poultry industry but also serious public health concerns. The full-length viral hemagglutinin (HA), neuraminidase (NA) or b...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/j.jviromet.2011.05.006
更新日期：2011-08-01 00:00:00
abstract：:Hepatitis B virus (HBV) is hyperendemic to southern Africa, with genotype A of HBV being the predominant genotype, and subgenotype A1 prevailing. Infection with this subgenotype is associated with rapid disease progression, and high frequency of hepatocellular carcinoma development. The objectives of our study was to ...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/j.jviromet.2014.03.015
更新日期：2014-07-01 00:00:00
abstract：:E(rns) is an envelope glycoprotein of classical swine fever virus (CSFV) with RNase activity. The purpose of this study was to produce an active E(rns) for further applications using the yeast secreted expression system. The E(rns) gene was cloned into the expression vector pGAPZalphaC which was introduced into Pichia...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/j.jviromet.2005.08.020
更新日期：2006-03-01 00:00:00
abstract：:Dried blood spots (DBS) expedite the collection, storage and shipping of blood samples, thereby facilitating large-scale serologic studies. We evaluated the sensitivity of anti-HCV IgG testing and HCV-RNA quantitation using freshly prepared and stored DBS derived from HCV-infected patients. Protocols for elution were ...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/j.jviromet.2014.10.018
更新日期：2015-02-01 00:00:00
abstract：:A real-time reverse transcription-polymerase chain reaction (qRT-PCR) assay was developed for the fast and highly sensitive detection of the sacbrood virus (SBV) genome and applied to honeybee samples. Using plasmid DNA containing a partial SBV genome and diluted serially, as few as 1×10(2)copies/μl (correlation co-ef...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/j.jviromet.2011.10.014
更新日期：2012-01-01 00:00:00
abstract：:Bovine viral diarrhea virus (BVDV) should be a ubiquitous viral pathogen to the cattle and sheep industry. This pathogen is responsible for severe economic losses. We previously showed that plasmid-mediated dual short hairpin RNA (shRNA) efficiently inhibit BVDV replication in bovine kidney epithelial (MDBK) cells. In...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/j.jviromet.2015.03.014
更新日期：2015-06-15 00:00:00
abstract：:Hepatitis A virus (HAV) harvested from infected MRC-5 cells can hemagglutinate various species of erythrocytes at acid pH (Eckels et al., 1989). Further studies revealed that the majority of the hemagglutinin (HA) in MRC-5 and BS-C-1 cells was cell-associated. A simplified procedure for preparing HAV-HA consisted of c...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/0166-0934(90)90123-w
更新日期：1990-06-01 00:00:00
abstract：:We have developed a sensitive method to detect and localize HPV-6 viral DNA, mRNA and protein in biopsy specimens of genital and respiratory tract lesions by using in situ hybridization and immunoperoxidase assays on sections of plastic-embedded tissue. This modified in situ hybridization technique, using ultrathin se...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/0166-0934(89)90098-0
更新日期：1989-07-01 00:00:00
abstract：:A method is described that allows the rapid screening of field isolates of rotavirus for the detection of specific viral RNA segments. Cloned cDNA copies of viral genomic RNAs are employed for detection in the assay which makes use of the dot-hybridization technique of Thomas (1980). The assay developed was shown to b...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/0166-0934(84)90009-0
更新日期：1984-10-01 00:00:00
abstract：:An H5N1 avian influenza virus (AIV) hemagglutinin (HA) protein pseudotyped lentivirus, HIV/H5-HA, was generated, characterized in vitro and evaluated for its ability to induce protective immunity against virulent wild type AIV in mice. The HIV/H5-HA virus was able to infect 293T, BHK, Vero, PK-15, MDCK cells but not I...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/j.jviromet.2008.08.016
更新日期：2008-12-01 00:00:00
abstract：:The aim of this study was the development of gag and pol dual labelled probe real-time PCR and RT PCR assays to quantify the proviral load and the transcripts of the British Visna/maedi virus EV1 strain. Primers and probes were chosen based on the consensus sequences of gag and pol clones representative of EV1 genetic...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/j.jviromet.2010.01.013
更新日期：2010-05-01 00:00:00
abstract：:An immunocapture ELISA test for the diagnosis of human and animal influenza A and/or B is described. A monoclonal anti-nucleoprotein (NP) antibody was used to capture the NP antigen and the captured antigen was detected by an anti-NP polyclonal rabbit antiserum. Compared with the usual diagnostic method by cultivation...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/0166-0934(89)90102-x
更新日期：1989-07-01 00:00:00
abstract：:Foot-and-mouth disease is one of the most economically important virus diseases of livestock. Two important requirements for the control of this disease are rapid laboratory diagnosis and epidemiological investigation. The use of the polymerase chain reaction method (PCR) to amplify specific nucleic acid regions offer...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/0166-0934(93)90176-r
更新日期：1993-04-01 00:00:00
abstract：:Hyperimmune rat antisera prepared against 5 recent antigenic variants of influenza A (H3N2) viruses were studied for haemagglutination inhibiting (HI) antibodies to the homologous and the heterologous viruses. The ratios of homologous to heterologous reactions varied from one animal to another in immunizations with ea...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/0166-0934(87)90147-9
更新日期：1987-03-01 00:00:00
abstract：:Viruses belonging to the Flaviviridae family are found and distributed in most of the tropical and sub-tropical regions of the world. The genus has more than 56 members, most of which cause clinical symptoms in humans. The clinical diagnosis of dengue requires laboratory confirmation because of the similarity of sympt...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/j.jviromet.2012.08.006
更新日期：2012-12-01 00:00:00
abstract：:Cellular nucleic acids can interfere with the molecular cloning of retroviruses, a problem that is particularly serious with viruses propagated in lymphoblastoid cells that release large amounts of microvesicles and other cellular components. The approach taken to circumvent such problems involved first suspending vir...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/j.jviromet.2006.07.003
更新日期：2006-11-01 00:00:00
abstract：:A method for coupling cytomegalovirus (CMV) antigens to aldehyde-fixed erythrocytes for use in a passive hemagglutination assay for antibody to CMV is described. This method uses acid buffer instead of tannic acid for coupling CMV-complement fixation (CF) antigen to the stabilized cells. The coated cells are stable fo...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/0166-0934(83)90002-2
更新日期：1983-09-01 00:00:00
abstract：:Rift Valley fever virus (RVFV) is an arthropod-borne pathogen that often results in severe morbidity and mortality in both humans and livestock. As its geographic range continues to expand, it presents a real threat to naïve populations around the world by accidental introduction (e.g., the result of increased travel)...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/j.jviromet.2010.07.015
更新日期：2010-11-01 00:00:00
