scholarly journals Partial characterization of Prunus Necrotic Ringspot Virus on apple in EGYPT 

2021 ◽  
Author(s):  
Aly M. Abdel-Salam ◽  
Samah A. Mokbel
Keyword(s):  
Binding Assay ◽  
Ringspot Virus ◽  
Economic Losses ◽  
Rt Pcr ◽  
Chain Reaction ◽  
Prunus Necrotic Ringspot Virus ◽  
Polymerase Chain ◽  
Severe Isolate ◽  
Microscopy Examination

Abstract A severe isolate of Prunus necrotic ringspot virus (PNRSV) was isolated from apple orchards in the vicinity of Nubaria city, Beheira governorate, Egypt. Infected-apple trees showed chlorotic, necrotic ringspots, and shoot holes on leaves. Severely infected- trees withered, became useless, and were removed causing severe economic losses. Reverse transcriptase (RT) polymerase chain reaction (PCR), RT-PCR, using degenerate primer pair for the coat protein (CP) gene of Ilarvirus amplified products similar to those produced from peach and apricot isolates of PNRSV-infecting stone fruits). Dot blotting immuno-binding assay (DBIA showed a positive reaction between PNRSV-infected apple sap and an Egyptian antiserum for PNRSV. Purified preparation from infected leaves, using the electro-elution technique yielded nucleoprotein, which had Amax and Amin at 260 and 240 nm respectively. Electron microscopy examination showed spherical virions with ca. 26 nm in diameter.

scholarly journals Karakterisasi Molekuler Papaya ringspot virus tipe P pada Tanaman Mentimun di Jawa

2018 ◽  
Vol 14 (3) ◽  
pp. 75
Author(s):  
Listihani Listihani ◽  
Tri Asmira Damayanti ◽  
Sri Hendrastuti Hidayat ◽  
Suryo Wiyono
Keyword(s):  
Ringspot Virus ◽  
Mosaic Symptom ◽  
Papaya Ringspot Virus ◽  
Rt Pcr ◽  
Chain Reaction ◽  
Specific Primers ◽  
Central Java ◽  
Dna Fragment ◽  
Polymerase Chain

Moleculer Characterization of Papaya ringspot virus type P on Cucumber in JavaInfection of Papaya ringspot virus (PRSV) on cucumber plants showing mosaic symptom was detected using specific antibody.  Further investigation was conducted to determine molecular characters and status of PRSV infecting cucumber in Java.  Infection of PRSV was detected from leaf samples collected from the field using dot immunobinding assay (DIBA).  Disease frequency caused by PRSV infection reached 81.11%, 95.86%, 91.66%, and 92.3% in East Java, Central Java, Yogyakarta, and West Java, respectively.  Characterization of PRSV isolates was conducted by reverse transcription polymerase chain reaction (RT-PCR) using specific primers for PRSV-P and PRSV-W, followed by cloning, and DNA sequencing.  DNA fragment of 470 bp was successfully amplified using specific primers for PRSV-P from several samples from Nganjuk, Brebes, Kulon Progo, and Subang; but no amplification was achieved using specific primers for PRSV-W.  Nucleotide and amino acid analysis showed high homology among PRSV-P isolates from Nganjuk, Brebes, Kulon Progo, and Subang, i.e. 98.6%-99.7% and 99.3%-100%, respectively.  This is an indication of a low genetic variation among PRSV-P from Java. Further phylogenetic analysis indicated that PRSV-P isolate cucumber is in the same cluster with PRSV-P isolate papaya from Bali, Indonesia.  This is the first report of PRSV-P infecting cucumber in Indonesia.

scholarly journals Sensitive SYBR Green—Real Time PCR for the Detection and Quantitation of Avian Rotavirus A

2018 ◽  
Vol 6 (1) ◽  
pp. 2 ◽  
Author(s):  
David De la Torre ◽  
Claudete Astolfi-Ferreira ◽  
Ruy Chacon ◽  
Antonio Piantino Ferreira
Keyword(s):  
Polymerase Chain Reaction ◽  
Real Time ◽  
Sybr Green ◽  
Molecular Techniques ◽  
Economic Losses ◽  
Rt Pcr ◽  
Chain Reaction ◽  
Rotavirus A ◽  
Avian Nephritis Virus ◽  
Polymerase Chain

Avian rotavirus A (ARtV-A) is a virus that affects young birds, causing acute diarrhea and economic losses in the poultry industry worldwide. The techniques used for the diagnosis of ARtV-A include electron microscopy, isolation in cell culture, and serology, as well as molecular techniques, such as the reverse transcription-polymerase chain reaction (RT-PCR). The objective of this work was to standardize a real-time RT-polymerase chain reaction (RT-qPCR) using SYBR Green chemistry for the rapid detection and quantification of ARtV-A from bird tissues and materials fixed on FTA cards on the basis of the nucleotide sequence of segment 6 (S6), which codes for the structural VP6 protein of ARtV-A. The results show the efficient amplification of the proposed target, with a limit of detection (LoD) of one copy gene (CG) per microliter of cDNA and a limit of quantification (LoQ) of 10 CGs per microliter. The efficiency of the primers was determined to be 95.66% using a standard curve, with an R2 value of 0.999 and a slope of −3.43. The specificity was determined using samples coinfected with ARtV-A, the chicken parvovirus, the chicken astrovirus, and the avian nephritis virus as positive controls and commercially available vaccines of the infectious bronchitis virus, infectious bursa disease virus, avian reovirus and healthy organs as negative controls. This technique, which lacks nonspecific PCR products and dimers, demonstrated greater sensitivity and specificity than conventional RT-PCR, and it reduced the analysis time by more than 50%.

scholarly journals First Detection and Characterization of Chrysanthemum virus B infecting Chrysanthemum in Thailand

2021 ◽  
Author(s):  
Salit Supakitthanakorn ◽  
Garnjana Wichitrakoonthavorn ◽  
Kaewalin Kunasakdakul ◽  
On-Uma Ruangwong
Keyword(s):  
Cultural Values ◽  
Ornamental Plants ◽  
Systemic Symptom ◽  
Rt Pcr ◽  
Chain Reaction ◽  
Transmission Electron ◽  
Staining Methods ◽  
Polymerase Chain ◽  
Systemic Symptoms

Abstract Chrysanthemum is one of the important ornamental plants in worldwide due to its high economic and cultural values. Chrysanthemum leaves showed mosaic, ringspot, yellowing and mild mottle symptoms were observed and collected from cultivation areas in northern Thailand and used for detection of important viruses infecting chrysanthemum. Chrysanthemum virus B (CVB) was detected by reverse transcription polymerase chain reaction (RT-PCR) from samples showing yellowing and mild mottle symptoms. Sequences of the coat protein (CP) gene of two CVB isolates found in this study were sequenced and shared 93.15% homology with other CVB isolates from different countries deposited in GenBank. Biological indexing of these CVB found that they induced both local and systemic symptoms in tobacco plants while petunia displayed a systemic symptom. The particles of CVB were observed under transmission electron microscope (TEM), prepared by dip preparation and negative staining methods, showing slightly flexuous rod-shaped virions approximately 600–650 nm in length. To our knowledge, this is the first detection and study on molecular and biological characteristics of CVB infecting chrysanthemum in Thailand.

scholarly journals Detection and molecular characterization of the Iris severe mosaic virus-Ir isolate from Iran

2015 ◽  
Vol 55 (3) ◽  
pp. 235-240 ◽  
Author(s):  
Masoud Nateqi ◽  
Mina Koohi Habibi ◽  
Akbar Dizadji ◽  
Shirin Parizad
Keyword(s):  
Amino Acid ◽  
Amino Acid Sequence ◽  
Mosaic Virus ◽  
Ornamental Plants ◽  
Rt Pcr ◽  
Chain Reaction ◽  
C Terminus ◽  
Coated Plate ◽  
Polymerase Chain

AbstractIris belongs to the Iridaceae family. It is one of the most important pharmaceutical and ornamental plants in the world. To assess the potyvirus incidence in natural resources of iris plants in Iran, Antigen Coated-Plate ELISA (ACP-ELISA) was performed on 490 symptomatic rhizomatous iris leaf samples, which detected the potyvirus in 36.7% of the samples. Genomic 3′ end of one mechanically non-transmitted potyvirus isolate, comprising a 3′ untranslated region (390 bp) and C-terminus of the coat protein (CP) gene (459 bp), was amplified by reverse transcription polymerase chain reaction (RT-PCR), which was ligated into pTG19-T vector. The nucleotide sequence of amplicons was compared with related sequences, using Blastn software available at NCBI GenBank, and showed the highest similarity withIris severe mosaic virus(ISMV) isolates. The nucleotide and deduced amino acid sequence of the CP C-terminus region was more than 83% identical with other ISMV isolates, therefore this isolate was designated as ISMV-Ir. This new ISMV isolate is closely related to the Chinese ISMV-PHz in phylogenetic analysis, based on the partial nucleotide and deduced amino acid sequence of the CP region. This is the first report of ISMV occurrence onIrisspp. in Iran.

scholarly journals Evidence for airborne transmission of Norwalk-like virus (NLV) in a hotel restaurant

2000 ◽  
Vol 124 (3) ◽  
pp. 481-487 ◽  
Author(s):  
P. J. MARKS ◽  
I. B. VIPOND ◽  
D. CARLISLE ◽  
D. DEAKIN ◽  
R. E. FEY ◽  
...  
Keyword(s):  
Electron Microscopy ◽  
Polymerase Chain Reaction ◽  
Inverse Relationship ◽  
Nucleotide Sequence Analysis ◽  
Rt Pcr ◽  
Chain Reaction ◽  
Virus Particles ◽  
Stool Samples ◽  
Polymerase Chain

An outbreak of gastroenteritis followed a meal in a large hotel during which one of the diners vomited. The clinical features of the illness suggested Norwalk-like virus (NLV, small round structured virus) infection, and this was confirmed by electron microscopy and reverse transcriptase polymerase chain reaction (RT–PCR) of stool samples. Further characterization of the virus by nucleotide sequence analysis of the PCR amplicons revealed identical strains in all the affected individuals. The foods served at the meal could not be demonstrated to be the cause of the outbreak. Analysis of attack rates by dining table showed an inverse relationship with the distance from the person who vomited. No one eating in a separate restaurant reported illness. Transmission from person-to-person or direct contamination of food seems unlikely in this outbreak. However, the findings are consistent with airborne spread of NLV with infection by inhalation with subsequent ingestion of virus particles.

Improved detection of prunus necrotic ringspot virus by the polymerase chain reaction

1996 ◽  
Vol 102 (7) ◽  
pp. 681-685 ◽  
Author(s):  
S. Spiegel ◽  
S. W. Scott ◽  
V. Bowman-Vance ◽  
Y. Tam ◽  
N. N. Galiakparov ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Ringspot Virus ◽  
Chain Reaction ◽  
Prunus Necrotic Ringspot Virus ◽  
Polymerase Chain
Sign in / Sign up

Export Citation Format

Share Document