当前位置： SCI文献检索 > JOURNAL OF VIROLOGICAL METHODS期刊下所有文献 > A real-time RT-PCR assay for detection and absolute quantitation of Citrus tristeza virus in different plant tissues.

A real-time RT-PCR assay for detection and absolute quantitation of Citrus tristeza virus in different plant tissues.

Abstract：

:A real-time RT-PCR assay using SYBR Green was developed for specific and reliable quantitative detection of Citrus tristeza virus (CTV) in infected plants. A general primer set designed from conserved sequences in ORFs 1b and 2 enabled amplification of the genomic RNA (gRNA) while excluding most subgenomic and defective RNAs. Single RT-PCR products of 204 bp (isolate T36) or 186 bp (other isolates) were obtained with no primer-dimer or non-specific amplifications detected. Melting curve analysis revealed distinct melting temperature peaks (T(m)) for severe and mild isolates. External standard curves using RNA transcripts of the selected target allowed a reproducible quantitative assay, with a wide dynamic range of detection starting with 10(2) gRNA copies and with very low variation coefficient values. This protocol enabled reliable assessments of CTV accumulation in different tissues and from different citrus species, grown in the greenhouse or under field conditions, and infected with CTV isolates differing in their pathogenicity. CTV accumulation was higher in bark and fruits than in roots or leaves and showed minimal differences among several susceptible citrus species, but it was significantly lower in sour orange. This quantitative detection assay will be a valuable tool for diagnosis and molecular studies on CTV biology.

journal_name

J Virol Methods

journal_title

Journal of virological methods

authors

Ruiz-Ruiz S,Moreno P,Guerri J,Ambrós S

doi

10.1016/j.jviromet.2007.05.011

subject

Has Abstract

pub_date

2007-11-01 00:00:00

pages

96-105

issue

2

eissn

0166-0934

issn

1879-0984

pii

S0166-0934(07)00178-4

journal_volume

145

pub_type

杂志文章

相关文献

JOURNAL OF VIROLOGICAL METHODS文献大全
  • Generation of a replication-deficient recombinant human adenovirus type 35 vector using bacteria-mediated homologous recombination.

    abstract：:The use of adenovirus type 35 (Ad35) as a vector in vaccine and gene therapy studies is promising due to its broad cell tropism and low seroprevalence in humans. However, to date, a simple and effective system for producing recombinant Ad35 (rAd35) has not been well developed. This report describes a two-plasmid Ad35-...

    journal_title：Journal of virological methods

    pub_type： 杂志文章

    doi：10.1016/j.jviromet.2011.06.016

    authors： Wu C,Lei X,Wang J,Hung T

    更新日期：2011-10-01 00:00:00

  • Methods for effective real-time RT-PCR analysis of virus-induced gene silencing.

    abstract：:We applied real-time RT-PCR to the analysis of Tobacco rattle virus (TRV)-mediated virus-induced gene silencing (VIGS) of the phytoene desaturase (PDS) gene in Nicotiana benthamiana and tomato. Using a combination of direct measurement and mathematical assessment, we evaluated three plant genes, ubiquitin (ubi3), elon...

    journal_title：Journal of virological methods

    pub_type： 杂志文章

    doi：10.1016/j.jviromet.2006.07.017

    authors： Rotenberg D,Thompson TS,German TL,Willis DK

    更新日期：2006-12-01 00:00:00

  • A cornucopia of screening and diagnostic techniques for human papillomavirus associated cervical carcinomas.

    abstract：:Cervical carcinoma is one of the major consequences of human papillomavirus (HPV) infections. Although HPV infections of cervix do not always progress to cancer, 90% cases of cervical cancer have been found associated with high risk HPV (hrHPV) infection. Usually, HPV infection is asymptomatic; however, this asymptoma...

    journal_title：Journal of virological methods

    pub_type： 杂志文章,评审

    doi：10.1016/j.jviromet.2015.06.015

    authors： Ramzan M,Noor ul Ain,Ilyas S,Umer M,Bano S,Sarwar S,Shahzad N,Shakoori AR

    更新日期：2015-09-15 00:00:00

  • Evaluation of enzyme-linked immunosorbent assays: a method of data analysis.

    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

    authors： Crofts N,Maskill W,Gust ID

    更新日期：1988-10-01 00:00:00

  • Establishment of a sensitive and specific hyper-branched rolling circle amplification assay and test strip for TSV.

    abstract：:A specific padlock probe (PLP) and detection probe were designed to target the capsid protein gene of Taura syndrome virus (TSV), and a hyper-branched rolling circle amplification (HRCA) assay and a corresponding strip-based test were produced. The reaction time and temperature were optimized for both. The PLPs were l...

    journal_title：Journal of virological methods

    pub_type： 杂志文章

    doi：10.1016/j.jviromet.2014.08.019

    authors： Zhao Y,Yin W,Wang Y,Wang G,Li B

    更新日期：2014-12-01 00:00:00

  • Identification of simian varicella virus gene 21 promoter region using green fluorescent protein.

    abstract：:Clinical, pathological, immunological and virological features of simian varicella virus (SVV) infection in primates closely resemble those of varicella zoster virus (VZV) infection in humans. In ganglia infected latently of humans and monkeys, gene 21 of VZV and SVV is transcribed, respectively. We determined the nuc...

    journal_title：Journal of virological methods

    pub_type： 杂志文章

    doi：10.1016/s0166-0934(99)00180-9

    authors： Mahalingam R,Wellish M,White T,Gilden DH

    更新日期：2000-04-01 00:00:00

  • Development of an epitope-blocking-enzyme-linked immunosorbent assay to differentiate between animals infected with and vaccinated against foot-and-mouth disease virus.

    abstract：:An epitope-blocking ELISA (EB-ELISA) was developed to distinguish animals infected with foot-and-mouth-disease (FMDV) from those immunized with commercial vaccines. The assay used monoclonal antibodies to target the 3B core repeat motif (QKPLK) and purified recombinant 3AB proteins from the major B cell line epitopes ...

    journal_title：Journal of virological methods

    pub_type： 杂志文章

    doi：10.1016/j.jviromet.2007.01.025

    authors： Oem JK,Chang BS,Joo HD,Yang MY,Kim GJ,Park JY,Ko YJ,Kim YJ,Park JH,Joo YS

    更新日期：2007-06-01 00:00:00

  • PG-4 cell plaque assay for xenotropic murine leukemia virus.

    abstract：:Xenotropic murine leukemia virus (X-MuLV) is often used in retrovirus elimination studies involving rodent cells. Currently, X-MuLV is measured using a focus-forming assay on mink (MiCl1 S+L-) or cat (PG-4 S+L-) cell lines. An easier and quicker PG-4 cell plaque assay, which retains the statistical reproducibility of ...

    journal_title：Journal of virological methods

    pub_type： 杂志文章

    doi：10.1016/s0166-0934(99)00064-6

    authors： Li Z,Blair M,Thorner L

    更新日期：1999-08-01 00:00:00

  • Chemical inhibition of foamyvirus in primary baboon (Papio cynocephalus) kidney cells.

    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

    authors： Weiss SA,Heberling RL,Kalter SS

    更新日期：1982-05-01 00:00:00

  • Infectious entry of reovirus cores into mammalian cells enhanced by transfection.

    abstract：:The reovirus virion is a moderately complex structure that contains eight structural proteins organized in multiple concentric capsid layers. Mammalian orthoreovirus virions undergo partial uncoating to produce infectious subvirion particles (ISVPs) and cores. Virions and ISVPs are infectious, whereas cores are transc...

    journal_title：Journal of virological methods

    pub_type： 杂志文章

    doi：10.1016/j.jviromet.2005.04.005

    authors： Jiang J,Coombs KM

    更新日期：2005-09-01 00:00:00

  • In vitro replication of Macrobrachium rosenbergii nodavirus and extra small virus in C6/36 mosquito cell line.

    abstract：:White tail disease (WTD) is a serious problem in hatcheries and nursery ponds of Macrobrachium rosenbergii in India and many parts of the world. The pathogenic agents have been identified as M. rosenbergii nodavirus (MrNV) associated with extra small virus (XSV), which is 27nm and 15nm in diameter, respectively. Repli...

    journal_title：Journal of virological methods

    pub_type： 杂志文章

    doi：10.1016/j.jviromet.2007.06.011

    authors： Sudhakaran R,Parameswaran V,Sahul Hameed AS

    更新日期：2007-12-01 00:00:00

  • Characterization of isolates of Citrus tristeza virus by sequential analyses of enzyme immunoassays and capillary electrophoresis-single-strand conformation polymorphisms.

    abstract：:Citrus tristeza virus (CTV) is the causal agent of tristeza disease, which is one of the most devastating diseases of citrus crops worldwide. This paper describes a method for the rapid detection and genotyping of naturally spreading CTV isolates. This method uses ELISA or dot-blot immunological tests to detect trees ...

    journal_title：Journal of virological methods

    pub_type： 杂志文章

    doi：10.1016/j.jviromet.2012.01.015

    authors： Licciardello G,Raspagliesi D,Bar-Joseph M,Catara A

    更新日期：2012-05-01 00:00:00

  • Freeze-drying foot-and-mouth disease virus antigens. II. For use in the ELISA.

    abstract：:Live and inactivated preparations of foot-and-mouth disease virus strains 01 BFS 1860 and A22 IRQ 24/64 were freeze-dried in the presence or absence of additive solutions and assessed for their reactivity by ELISA at intervals over a six month storage period at various temperatures and also after reconstitution and su...

    journal_title：Journal of virological methods

    pub_type： 杂志文章

    doi：10.1016/0166-0934(90)90019-c

    authors： Ferris NP,Philpot RM,Oxtoby JM,Armstrong RM

    更新日期：1990-11-01 00:00:00

  • Development of reverse transcription loop-mediated isothermal amplification assay for rapid detection of an emerging potyvirus: tomato necrotic stunt virus.

    abstract：:Tomato necrotic stunt virus (ToNStV) is an emerging potyvirus that causes severe stunting to infected tomato plants. A reverse transcription loop-mediated isothermal amplification (RT-LAMP) assay was developed for sensitive detection of ToNStV. The sensitivity of RT-LAMP was comparable to that of conventional RT-PCR, ...

    journal_title：Journal of virological methods

    pub_type： 杂志文章

    doi：10.1016/j.jviromet.2014.01.017

    authors： Li R,Ling KS

    更新日期：2014-05-01 00:00:00

  • A flow-through cell counting assay for point-of-care enumeration of CD4 T-cells.

    abstract：:CD4 T-cell count is a priority for staging HIV disease and guiding clinical management as part of HIV care. Conventional CD4 T-cell enumeration methods based on flow cytometry are expensive, require well-trained personnel, and are challenging to use in rural, resource-scarce areas. A simple CD4 T-cell count test that ...

    journal_title：Journal of virological methods

    pub_type： 杂志文章

    doi：10.1016/j.jviromet.2019.05.012

    authors： Bystryak S,Bandwar RP,Santockyte R

    更新日期：2019-09-01 00:00:00

  • Quantitative real-time PCR in paediatric patients with herpes simplex infections of the central nervous system.

    abstract：:The aim of this study was to evaluate viral DNA load in paediatric patients with herpes simplex infections of the central nervous system. A real-time PCR assay for herpes simplex types 1 and 2 was developed for this 8-year retrospective study that included children with herpes simplex infection of the central nervous ...

    journal_title：Journal of virological methods

    pub_type： 杂志文章

    doi：10.1016/j.jviromet.2007.09.009

    authors： Muñoz-Almagro C,Jordan I,Cambra FJ,Esteban E,Urrea M,Garcia-Garcia JJ,Palomeque A

    更新日期：2008-02-01 00:00:00

  • Physical characterization of PBCV virus by sedimentation field flow fractionation.

    abstract：:A virus (PBCV) from the symbiotic Chlorella-like algae of Paramecium bursaria has been characterized by sedimentation field flow fractionation (SdFFF). The method yields effective molecular weight M' (molecular weight X buoyancy factor) with a precision of better than 5% under a variety of experimental conditions. The...

    journal_title：Journal of virological methods

    pub_type： 杂志文章

    doi：10.1016/0166-0934(85)90038-2

    authors： Yonker CR,Caldwell KD,Giddings JC,Van Etten JL

    更新日期：1985-06-01 00:00:00

  • Development of a recombinant ELISA using yeast (Pichia pastoris)-expressed polypeptides for detection of antibodies against avian influenza A subtype H5.

    abstract：:Two truncated sequences (designated P1 and rHA1) of influenza A virus subtype H5 haemagglutinin (HA) were cloned and expressed in yeast Pichia pastoris (P. pastoris). These polypeptides were used in an indirect recombinant ELISA (rELISA) for detection of H5 antibodies in poultry. Serum samples obtained from broiler ch...

    journal_title：Journal of virological methods

    pub_type： 杂志文章

    doi：10.1016/j.jviromet.2011.12.004

    authors： Shehata AA,Fiebig P,Sultan H,Hafez M,Liebert UG

    更新日期：2012-03-01 00:00:00

  • A synthetic parvovirus B19 capsid protein can replace viral antigen in antibody-capture enzyme immunoassays.

    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

    authors： Kock WC

    更新日期：1995-09-01 00:00:00

  • Quantivirus® HPV E6/E7 RNA 3.0 assay (bDNA) is as sensitive, but less specific than Hybrid Capture 2 test.

    abstract：:Human papillomavirus (HPV) infection is the primary cause of cervical cancer. The Quantivirus(®) HPV E6/E7 RNA 3.0 assay (DiaCarta, CA, USA) detects E6/E7 mRNA of 13 high risk subtypes and 6 low risk subtypes. Cervical specimens collected in PreservCyt were processed for HPV detection. Cervical biopsies were taken onl...

    journal_title：Journal of virological methods

    pub_type： 杂志文章

    doi：10.1016/j.jviromet.2012.11.024

    authors： Shen Y,Gong J,He Y,Cheng G,Okunieff P,Li X

    更新日期：2013-02-01 00:00:00

  • Detection of human virulence signatures in H5N1.

    abstract：:A method for detecting the emergence of potential pandemic-causing influenza strains has been developed. The system first uses real-time RT-PCR to detect H5, the highly pathogenic avian influenza subtype most likely to cause a pandemic. Pyrosequencing is then employed to scan for codon changes encoding amino acids kno...

    journal_title：Journal of virological methods

    pub_type： 杂志文章

    doi：10.1016/j.jviromet.2008.09.013

    authors： Waybright N,Petrangelo E,Lowary P,Bogan J,Mulholland N

    更新日期：2008-12-01 00:00:00

  • An in-well direct lysis method for rapid detection of SARS-CoV-2 by real time RT-PCR in eSwab specimens.

    abstract：BACKGROUND:Diagnostic real time reverse transcription PCR (rRT-PCR) is usually done using nucleic acid (NA) purified from the sample. In the SARS-CoV-2 pandemic reagents and utensils for NA purification has been in short supply. This has generated interest in methods that eliminate the need for NA purification. OBJECT...

    journal_title：Journal of virological methods

    pub_type： 杂志文章

    doi：10.1016/j.jviromet.2021.114062

    authors： Jørgensen RL,Pedersen MS,Chauhan AS,Andreasson LM,Kristiansen GQ,Lisby JG,Rosenstierne MW,Schønning K

    更新日期：2021-01-08 00:00:00

  • Diagnosis of canine coronavirus infection using nested-PCR.

    abstract：:The results of polymerase chain reaction (PCR) and nested polymerase chain reaction (n-PCR) assays for the diagnosis of canine coronavirus (CCV) infection, and the comparison with other diagnostic techniques, such as electron microscopy (EM) and virus isolation using A-72 cell line are reported. The study was carried ...

    journal_title：Journal of virological methods

    pub_type： 杂志文章

    doi：10.1016/s0166-0934(99)00134-2

    authors： Pratelli A,Buonavoglia D,Martella V,Tempesta M,Lavazza A,Buonavoglia C

    更新日期：2000-01-01 00:00:00

  • A loop-mediated isothermal amplification coupling with a lateral flow dipstick for rapid and specific detection of fowl adenovirus serotype-4.

    abstract：:Fowl adenovirus serotype-4 (FAdV-4) has been recognized as a predominant threat to the broilers aged from three to five weeks. Hydropericardium syndrome (HPS) is one of its major clinical diseases by FAdV-4 resulting in heavy economic losses. In this study, a loop-mediated isothermal amplification coupling with a late...

    journal_title：Journal of virological methods

    pub_type： 杂志文章

    doi：10.1016/j.jviromet.2019.04.026

    authors： Zhai X,Mei X,Wu X,Zuo L,Zhou L,Tian Y,Han X,Yang X,Wang H

    更新日期：2019-08-01 00:00:00

  • Development and application of antibody microarray for lymphocystis disease virus detection in fish.

    abstract：:Lymphocystis disease virus (LCDV) is the causative agent of lymphocystis disease affecting marine and freshwater fish worldwide. Here an antibody microarray was developed and employed to detect LCDV in fish. Rabbit anti-LCDV serum was arrayed on agarose gel-modified slides as capture antibody, and Cy3-conjugated anti-...

    journal_title：Journal of virological methods

    pub_type： 杂志文章

    doi：10.1016/j.jviromet.2013.02.015

    authors： Sheng X,Xu X,Zhan W

    更新日期：2013-05-01 00:00:00

  • Low abundance drug resistance variants in transmitted HIV drug resistance surveillance specimens identified using tagged pooled pyrosequencing.

    abstract：:HIV drug resistance (DR) testing using Sanger sequencing (SS) is limited by the inability of the method to identify low abundance drug resistance variants. The application of tagged pooled pyrosequencing (TPP) for HIV DR surveillance is described and the results compared with SS. HIV(+) serum specimens were genotyped ...

    journal_title：Journal of virological methods

    pub_type： 杂志文章

    doi：10.1016/j.jviromet.2012.10.018

    authors： Ji H,Liang B,Li Y,Van Domselaar G,Graham M,Tyler S,Merks H,Sandstrom P,Brooks J

    更新日期：2013-02-01 00:00:00

  • Half-genome human immunodeficiency virus type 1 constructs for rapid production of reporter viruses.

    abstract：:The use of reporter constructs of HIV-1 molecular clones has been useful in studying the effects of infection on individual cells. A strategy of producing viral half-genome constructs containing reporter genes that can be used to produce recombinant complete infectious virions is described. This approach allows rapid ...

    journal_title：Journal of virological methods

    pub_type： 杂志文章

    doi：10.1016/s0166-0934(03)00110-1

    authors： Ali A,Jamieson BD,Yang OO

    更新日期：2003-06-30 00:00:00

  • Quantitative and sensitive detection of immune-complexed and free HIV antigen after boiling of serum.

    abstract：:5-min boiling of 3-fold diluted serum leads to the quantitative release of immune-complexed HIV antigen (Ag) that can be readily measured. The method is capable of detecting as little as 4 to 5 pg/ml Ag p24 depending on the Ag test used. Of a panel of 50 anti-HIV-positive sera, 30 (60%) were Ag-positive after heat den...

    journal_title：Journal of virological methods

    pub_type： 杂志文章

    doi：10.1016/0166-0934(93)90080-b

    authors： Schüpbach J,Böni J

    更新日期：1993-07-01 00:00:00

  • A non-phenol-chloroform extraction of double-stranded RNA from plant and fungal tissues.

    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

    authors： Balijja A,Kvarnheden A,Turchetti T

    更新日期：2008-09-01 00:00:00

  • The development of a loop-mediated isothermal amplification assay for rapid and sensitive detection of abalone herpesvirus DNA.

    abstract：:A loop-mediated isothermal amplification (LAMP) assay was developed for the detection of abalone herpesvirus DNA. Two pairs of primers were designed, based on the sequence of the DNA polymerase gene of abalone herpesvirus. The reaction temperature and time were optimized to 63°C and 60min, respectively. LAMP amplicons...

    journal_title：Journal of virological methods

    pub_type： 杂志文章

    doi：10.1016/j.jviromet.2013.11.011

    authors： Chen MH,Kuo ST,Renault T,Chang PH

    更新日期：2014-02-01 00:00:00