A broad spectrum, one-step RT-PCR to detect Satsuma dwarf virus variants using universal primers targeting both segmented RNAs 1 and 2

Shimizu, Saori; Ito, Takao; Miyoshi, Takanori; Tachibana, Yasunobu; Ito, Takumi

doi:10.1007/s10327-011-0342-x

Cited by 13 publications

(8 citation statements)

References 6 publications

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“…To check the graft transmissibility of the viral molecule, scions from Kaki13-14 were grafted to four 'Fuyu' seedlings. Reverse transcription (RT)-PCR with specific primers, the homologous 5RhF1 (59-GGATGGCTTCTGATGTGTGG-39, nt 7575-7594) and the complementary 5RhC1 (59-TTGA-CCCGAATATGTGCGAT-39, nt 7852-7871), were carried out using a OneStep RT-PCR kit (Qiagen) as described by Shimizu et al (2011) with some modifications. At 18 months post-inoculation, RT-PCR detected the molecule in all four grafted seedlings that had previously tested negative.…”

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confidence: 99%

Genetic characterization of novel putative rhabdovirus and dsRNA virus from Japanese persimmon

Ito

Suzaki

Nakano

2013

Journal of General Virology

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Deep-sequencing analysis of nucleic acids from leaf tissue of Japanese persimmon trees exhibiting fruit apex disorder in some fruits detected two molecules that were graft transmitted to healthy seedlings. One of the complete genomes consisted of 13 467 nt and encoded six genes similar to those of plant rhabdoviruses. The virus formed a distinct cluster in the genus Cytorhabdovirus with lettuce necrotic yellows virus, lettuce yellow mottle virus and strawberry crinkle virus in a phylogenetic tree based on the L protein (RNA-dependent RNA polymerase, RdRp). The other consisted of 7475 nt and shared a genome organization similar to those of some insect and fungal viruses having dsRNA genomes. In a phylogenetic tree using the RdRp sequence of several unassigned dsRNA viruses, the virus formed a possible new genus cluster with two insect viruses, Circulifer tenellus virus 1 and Spissistilus festinus virus 1, and one plant virus, cucurbit yellows-associated virus.

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Genetic characterization of novel putative rhabdovirus and dsRNA virus from Japanese persimmon

Ito

Suzaki

Nakano

2013

Journal of General Virology

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“…A total of 121 citrus leaf samples exhibiting some foliar symptoms (such as yellowing and mottling) and/or tree decline were randomly collected during 2016 to 2019 from orchards of six major citrus-growing provinces in China, including Chongqing, Guangxi, Jiangxi, Sichuan, Yunnan, and Zhejiang. SDV was detected by reverse-transcriptase (RT)-PCR using a universal primer pair of SDV (Table S1; Shimizu et al 2011), and a CiMV-infected Goutoucheng sour orange named GTC was used as the positive control (Zhou et al 1993a). Total RNA of leaf tissues was extracted using TRIzol® reagent (Thermo-Fisher Scientific, Waltham, USA) as per the manufacturer's directions.…”

Section: Electronic Supplementary Materialsmentioning

confidence: 99%

Complete genome sequences and recombination analysis of three divergent Satsuma dwarf virus isolates

Yan

Zhang

et al. 2020

Trop. plant pathol.

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Citrus mosaic virus (CiMV) and navel orange infectious mottling virus (NIMV) are considered strains of satsuma dwarf virus (SDV). To understand the distribution of different isolates of SDV in China, a field survey was conducted, which allowed for the detection of CiMV and NIMV in citrus plants in Zhejiang and Sichuan provinces, respectively. In the present work, two CiMV isolates, including one named CiMV-GTC1 from Goutoucheng sour orange and one named CiMV-SE1 from Setoka tangor, as well as one NIMV isolate named NIMV-EH1 from Ehime-ken No. 38 tangor were, for the first time, fully sequenced. Among the isolates, CiMV-SE1 and CiMV-GTC1 were closely associated with the partial genomes of known CiMV isolates, while NIMV-EH1 was markedly different from known NIMV isolate. Sequence analysis demonstrated that CiMV, NIMV, and SDV shared similar genome organization and high nucleotide identities at the genome level. In addition, phylogenetic reconstruction of the three viruses also suggested a close relationship. Furthermore, recombination analysis provided evidence for their common cooccurrence in nature. These findings contribute significantly to our understanding of the natural evolution of SDV populations and can provide a basis for preventing the spread of different SDV isolates in the main citrus production regions.

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“…Virus samples (CiMV Az-1(B291), SDV S-58, and NIMV NI-1) were prepared from leaves of cape lemon trees that virus infection is maintained in Fruit tree research center, Ehime research institute of agriculture, forestry, and fisheries. The existence of viruses in samples was confirmed by RT-PCR analysis according to previously described method [7]. As a negative control, we also prepared a virus-free sample from non-infected leaves.…”

Section: Enzyme-linked Immunosorbent Assaymentioning

confidence: 99%

“…CiMV belongs to a group of SDV-like viruses [21]. The group consists of Satsuma dwarf virus (SDV), CiMV, Navel orange infectious virus (NIMV), and Hyuganatsu virus (HV) [7]. To investigate the specificity of anti-CiMV mAbs isolated in this study, we analyzed cross reactivity of these mAbs with SDV-like viruses.…”

Section: Specificity Of Anti-cimv Mabsmentioning

confidence: 99%

“…All virions consist of two kinds of coat proteins (large component; 42 kDa and small component; 22 kDa) encoded by RNA2 [5,6]. SDV and SDV-related viruses have been diagnosed by polymerase chain reaction (PCR) and/or enzyme linked immunosorbent assay (ELISA) [1,7]. Since PCR and ELISA need specialized techniques and expensive equipment and reagents, an immunochromatographic assay (ICA) has been developed for detection of these viruses during field samples.…”

Section: Introductionmentioning

confidence: 99%

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Production of a rabbit monoclonal antibody for highly sensitive detection of citrus mosaic virus and related viruses

et al. 2020

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Citrus mosaic virus (CiMV) is one of the causal viruses of citrus mosaic disease in satsuma mandarins (Citrus unshiu). Prompt detection of trees infected with citrus mosaic disease is important for preventing the spread of this disease. Although rabbit monoclonal antibodies (mAbs) exhibit high specificity and affinity, their applicability is limited by technical difficulties associated with the hybridoma-based technology used for raising these mAbs. Here, we demonstrate a feasible CiMV detection system using a specific rabbit mAb against CiMV coat protein. A conserved peptide fragment of the small subunit of CiMV coat protein was designed and used to immunize rabbits. Antigen-specific antibody-producing cells were identified by the immunospot array assay on a chip method. After cloning of variable regions in heavy or light chain by RT-PCR from these cells, a gene set of 33 mAbs was constructed and these mAbs were produced using Expi293F cells. Screening with the AlphaScreen system revealed eight mAbs exhibiting strong interaction with the antigen peptide. From subsequent sequence analysis, they were grouped into three mAbs denoted as No. 4, 9, and 20. Surface plasmon resonance analysis demonstrated that the affinity of these mAbs for the antigen peptide ranged from 8.7 × 10 −10 to 5.5 × 10 −11 M. In addition to CiMV, mAb No. 9 and 20 could detect CiMV-related viruses in leaf extracts by ELISA. Further, mAb No. 20 showed a high sensitivity to CiMV and CiMV-related viruses, simply by dot blot analysis. The anti-CiMV rabbit mAbs obtained in this study are envisioned to be extremely useful for practical applications of CiMV detection, such as in a virus detection kit.

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A broad spectrum, one-step RT-PCR to detect Satsuma dwarf virus variants using universal primers targeting both segmented RNAs 1 and 2

Cited by 13 publications

References 6 publications

Genetic characterization of novel putative rhabdovirus and dsRNA virus from Japanese persimmon

Genetic characterization of novel putative rhabdovirus and dsRNA virus from Japanese persimmon

Complete genome sequences and recombination analysis of three divergent Satsuma dwarf virus isolates

Production of a rabbit monoclonal antibody for highly sensitive detection of citrus mosaic virus and related viruses

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