scholarly journals Detection of Cherry leaf roll virus and Strawberry latent ring spot virus by one-step RT-PCR

2009 ◽  
Vol 45 (No. 4) ◽  
pp. 140-143 ◽  
Author(s):  
S. Kumari
Keyword(s):  
Leaf Roll ◽  
Rt Pcr ◽  
Spot Virus ◽  
Cherry Leaf Roll Virus ◽  
One Step ◽  
Polymerase Chain ◽  
The One ◽  
Ring Spot ◽  
Screen House ◽  
Xiphinema Diversicaudatum

A one-step reverse transcription-polymerase chain reaction (RT-PCR) protocol was developed and used for the detection of <i>Cherry leaf roll virus</i> (CLRV) and <i>Strawberry latent ring spot virus</i> (SLRSV). The protocol was used to test infected screen house plants and also plants from orchards and vineyards where the vector (<i>Xiphinema diversicaudatum</i>) of SLRSV was detected from the soil. The one-step RT-PCR protocol is rapid and sensitive and has the potential to be used for the diagnosis of CLRV and SLRSV in routine diagnostic laboratories.

scholarly journals Development of a Real-Time RT-PCR SYBR Green Assay for Tomato ring spot virus in Grape

Plant Disease ◽  
2007 ◽  
Vol 91 (9) ◽  
pp. 1083-1088 ◽  
Author(s):  
Elwin L. Stewart ◽  
Xinshun Qu ◽  
Barrie E. Overton ◽  
Fred E. Gildow ◽  
Nancy G. Wenner ◽  
...  
Keyword(s):  
Real Time ◽  
Gene Sequence ◽  
Sybr Green ◽  
Enzyme Linked Immunosorbent Assay ◽  
Rt Pcr ◽  
Spot Virus ◽  
Sybr Green Assay ◽  
One Step ◽  
Polymerase Chain ◽  
Ring Spot

Grapevines infected with Tomato ring spot virus (ToRSV) pose an economic risk for growers in the northeastern United States. This study describes a one-step real-time reverse-transcription polymerase chain reaction (RT-PCR) SYBR Green assay for detecting ToRSV in grapevines. Two newly designed primer pairs based on the ToRSV coat protein gene sequence were evaluated for specificity and optimized for a SYBR Green assay. The primer pair ToRSV1f/1r yielded a 130-bp product with strong primer-dimer products, whereas the primer pair ToRSV2f/2r yielded a 330-bp product with weak primer dimer products. Real-time RT-PCR detected ToRSV in more naturally infected grapevines maintained in the greenhouse than did enzyme-linked immunosorbent assay. The nucleotide sequences of the fragments amplified from grapevine growing in Pennsylvania using real-time PCR were divergent from previously published sequences.

Detection of Strawberry latent ring spot virus in Leaves of Olive Trees in Italy using a One-Step RT-PCR

2002 ◽  
Vol 150 (11-12) ◽  
pp. 636-639 ◽  
Author(s):  
F. Faggioli ◽  
L. Ferretti ◽  
G. Pasquini ◽  
M. Barba
Keyword(s):  
Rt Pcr ◽  
Spot Virus ◽  
One Step ◽  
Olive Trees ◽  
Ring Spot

scholarly journals A One-Step Reverse Transcription-Polymerase Chain Reaction System for the Simultaneous Detection and Identification of Multiple Tospovirus Infections

2005 ◽  
Vol 95 (2) ◽  
pp. 166-171 ◽  
Author(s):  
Hiroyuki Uga ◽  
Shinya Tsuda
Keyword(s):  
Polymerase Chain Reaction ◽  
Reverse Transcription ◽  
Simultaneous Detection ◽  
Yellow Spot ◽  
Rt Pcr ◽  
Chain Reaction ◽  
Spot Virus ◽  
Detection And Identification ◽  
One Step ◽  
Polymerase Chain

A one-step reverse transcription-polymerase chain reaction (RT-PCR) method has been developed for the simultaneous detection and identification of multiple tospoviruses that infect plants. The RT-PCR system is composed of six primers in a single tube: a universal degenerate primer and five virus species-specific primers. Amplifications resulted in an 848-bp PCR product for Watermelon silver mottle virus, 709-bp for Tomato spotted wilt virus, 589-bp for Impatiens necrotic spot virus, 511-bp for Melon yellow spot virus, and a 459-bp amplicon for Iris yellow spot virus. This system enables the simultaneous detection of at least three types of tospovirus infections, in addition to their species identities, from five possible tospoviruses studied, on the basis of their S RNA combinations. This multiplex RT-PCR system was applied to the detection of tospovirus in ornamental crops cultivated in fields and shows potential for epidemiological studies.

Evaluation of Minimal Residual Disease Using Reverse-Transcription Polymerase Chain Reaction in t(8;21) Acute Myeloid Leukemia: A Multicenter Study of 51 Patients

2000 ◽  
Vol 18 (4) ◽  
pp. 788-788 ◽  
Author(s):  
F. Morschhauser ◽  
J.M. Cayuela ◽  
S. Martini ◽  
A. Baruchel ◽  
P. Rousselot ◽  
...  
Keyword(s):  
Myeloid Leukemia ◽  
Residual Disease ◽  
Rank Test ◽  
Rt Pcr ◽  
Log Rank Test ◽  
One Step ◽  
Polymerase Chain ◽  
The One ◽  
Pcr Techniques

PURPOSE: Most studies using various reverse-transcription polymerase chain reaction (RT-PCR) techniques reported that the detection of the AML1-ETO fusion transcript was a common finding in long-term complete remission (CR) in acute myeloid leukemia (AML) with t(8;21) translocation. However, larger prospective studies with interlaboratory quality control may be important to investigate more precisely the clinical usefulness of studying minimal residual disease with RT-PCR in t(8;21) AML. PATIENTS AND METHODS: We collected 223 marrow samples from 51 patients with t(8;21) AML diagnosed in five centers and tested all samples by two different RT-PCR techniques (a nested technique and a one-step technique, with a sensitivity of 10−6 and 10−5, respectively) in two different laboratories. RESULTS: Samples from 14 patients in long persistent CR (median follow-up duration, 112 months) were taken at least twice, and all were PCR-negative by both techniques. Samples were prospectively taken from 37 patients after achievement of first CR and/or second CR, before intensive consolidation treatment, and every 3 to 6 months after completion of therapy. Patients who converted to PCR negativity with the one-step technique (60%) or both techniques (48%) after CR achievement had a longer CR duration than those with persistently positive PCR results (two-sided log-rank test, P = .0001). Patients who became PCR-negative with the one-step technique before intensive consolidation (23%) had a lower relapse rate (11% v 72%) and a longer CR duration than those who remained persistently PCR-positive at that point (two-sided log-rank test, P = .0015). CONCLUSION: Patients with AML with t(8;21) in long-term remission were all PCR-negative. In prospectively studied patients, a good correlation was found between negative PCR results and absence of relapse. Early negative results with the one-step RT-PCR technique, before consolidation treatment, seemed to carry an especially good prognosis, suggesting that RT-PCR analysis could help in choosing the type of consolidation therapy in patients with t(8;21) AML.

Comparison of ELISA and RT-PCR for the detection of Prunus necrotic ring spot virus and prune dwarf virus in almond (Prunus dulcis)

2003 ◽  
Vol 114 (1) ◽  
pp. 65-69 ◽  
Author(s):  
Genet Mekuria ◽  
Sunita A. Ramesh ◽  
Evita Alberts ◽  
Terry Bertozzi ◽  
Michelle Wirthensohn ◽  
...  
Keyword(s):  
Prunus Dulcis ◽  
Dwarf Virus ◽  
Rt Pcr ◽  
Spot Virus ◽  
Prune Dwarf Virus ◽  
Ring Spot

scholarly journals Identification and Host Relations of Turnip ringspot virus, A Novel Comovirus from Ohio

Plant Disease ◽  
2007 ◽  
Vol 91 (10) ◽  
pp. 1212-1220 ◽  
Author(s):  
P. Rajakaruna ◽  
S. Khandekar ◽  
T. Meulia ◽  
S. M. Leisner
Keyword(s):  
Genomic Sequence ◽  
Microscopic Analysis ◽  
Unknown Origin ◽  
Ringspot Virus ◽  
Rt Pcr ◽  
Electron Microscopic ◽  
Polymerase Chain ◽  
Line Patterns ◽  
Ring Spot ◽  
Host Relations

Viruslike chlorotic ring spot symptoms and line patterns of unknown origin were observed on a greenhouse-grown turnip plant. The suspected virus was mechanically transmissible to plants in the Brassicaceae. Electron microscopic analysis revealed icosahedral particles approximately 28 nm in diameter. Reverse transcriptase–polymerase chain reaction (RT-PCR) analyses suggested that the pathogen is a comovirus, an observation that was confirmed by analysis of portions of the genomic sequence. This virus was provisionally named Turnip ringspot virus (TuRSV). Based on the RNA 1 sequence, TuRSV is most similar to Radish mosaic virus, another pathogen that infects members of the Brassicaceae. Arabidopsis thaliana is susceptible to TuRSV, and 12 out of the 23 ecotypes studied showed symptoms when inoculated with the virus. TuRSV induced a variety of responses on ecotypes from death to no infection. Some ecotypes showed one or two rounds of symptom display followed by recovery when inoculated with TuRSV. About half of the ecotypes (11/23) analyzed showed no symptoms when inoculated with TuRSV. Col-0 plants showed no symptoms, and infectious virus was not recovered from systemic leaves, although it could be detected by RT-PCR. Col-0 plants harboring mutations impairing the ethylene, jasmonic acid, or salicylic acid signaling pathways did not show symptoms when inoculated with TuRSV.

scholarly journals Detection of Apple Stem Grooving Virus in Dormant Apple Trees with Crude Extracts as Templates for One-Step RT-PCR

Plant Disease ◽  
1998 ◽  
Vol 82 (7) ◽  
pp. 785-790 ◽  
Author(s):  
Vera L. A. Marinho ◽  
J. Kummert ◽  
G. Rufflard ◽  
D. Colinet ◽  
P. Lepoivre
Keyword(s):  
Rt Pcr ◽  
Degenerate Primers ◽  
Apple Trees ◽  
Active Growth ◽  
Japanese Strain ◽  
Apple Stem Grooving Virus ◽  
Apple Stem ◽  
Crude Extracts ◽  
One Step ◽  
The One

Partial nucleotide sequences of amplification products obtained from four European apple stem grooving virus (ASGV) isolates using degenerate primers showed 80 to 85% similarity with the published ASGV sequence of a Japanese strain but 98 to 100% identities among themselves. Based on these sequences, two ASGV-specific primers (ASGV4F-ASGV4R) were designed to amplify a 574-bp fragment located in the putative viral RNA polymerase. With these primers, six European and five American ASGV isolates, maintained in herbaceous hosts (Chenopodium quinoa, Nicotiana glutinosa, and N. occidentalis) or in apple trees, were readily detected by reverse transcription-polymerase chain reaction (RT-PCR). Using these specific ASGV primers, dsRNA preparations have been shown to constitute good templates for reliable amplification of ASGV sequences from leaves and bark tissues of apple trees, both in a two-step RT-PCR protocol and in the one-step Titan One-Tube RT-PCR. System. Furthermore, the one-step RT-PCR system allowed a specific amplification of ASGV sequences directly from clarified crude extracts of leaves and bark tissues of apple trees during both active growth and the dormant season.

scholarly journals Identification et caractérisation des isolats d'orbivirus des îles de la Réunion et de la Martinique

2009 ◽  
Vol 62 (2-4) ◽  
pp. 149
Author(s):  
D. Vitour ◽  
Corinne Sailleau ◽  
Emmanuel Breard ◽  
Stéphan Zientara
Keyword(s):  
Sequence Analysis ◽  
Clinical Symptoms ◽  
Enzyme Linked Immunosorbent Assay ◽  
La Réunion ◽  
Rt Pcr ◽  
Polymerase Chain ◽  
The One ◽  
Almost All ◽  
Haemorrhagic Disease

At the beginning of 2009, bluetongue (BT)-like clinical symptoms were reported in cattle on the French island of La Réunion (Indian Ocean). One hundred and twenty-three cows were blood tested for the presence of BT and/or epizootic haemorrhagic disease virus (EHDV) ribonucleic acid (RNA) by group specific reverse transcriptase polymerase chain reaction (RT-PCR). EHDV RNA was detected in 111 samples (90.2%), among which five were also positive for BTV RNA. Sequence analysis of EHDV segment 7 revealed that this circulating strain seemed to be similar to the one isolated in 2003 (99.8% nucleotide identity). The determination of the nucleotide sequence of segment 2 is under investigation. The vironeutralization test (VNT), serotype-specific RT-PCR, as well as sequence analysis identified the isolated BTV strain as serotype 2. These data showed that an EHDV outbreak occurred over the last winter in La Réunion, and it was concomitant to circulation of BTV. Epidemic or enzootic features of both these viruses are not yet known. Since this outbreak, molecular and serological tools specific to EHDV have been or are being developed. Three years ago, 30 healthy head of cattle moved from Metropolitan France to the French Martinique Island (Caribbean Basin) and were distributed in four different farms. Animals were sampled (blood and serum) every 10 days until day 30 and tested for BTV infection by the enzyme-linked immunosorbent assay (ELISA) or RT-PCR assays. Unexpectedly, almost all animals became BTV positive within 20 days. Whenever possible, virus isolation on eggs and baby hamster kidney (BHK) cell cultures were performed. Interestingly, seven BT strains belonging to seven distinct serotypes (BTV-2, 9, 10, 17, 18, 22, 24) were isolated. The coding sequence of segments 7, 8, 9 and 10 of these seven serotypes was obtained, as well as a portion of segment 2. The phylogenetic analysis revealed an unprecedented divergence of these strains with other known BTV sequences.

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