Simultaneous detection and identification of four cherry viruses by two step multiplex RT-PCR with an internal control of plant nad5 mRNA

Noorani, Salik; Awasthi, Poorwa; Sharma, Manju; Ram, Raja; Zaidi, A. A.; Hallan, Vipin

doi:10.1016/j.jviromet.2013.05.006

Cited by 26 publications

(23 citation statements)

References 38 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Particularly, this technique has been used successfully for simultaneous detection and differentiation of the five seedborne legume viruses [35], three ilarviruses affecting stone fruit trees [27], five potato viruses and a viroid [8], six citrus viroids and apple stem grooving virus from citrus plants [36], four viruses and Pseudomonas savastanoi pv. savastanoi in olive trees [10], five tospoviruses in ornamental crops [28], eight stone fruit viruses [37], nine grapevine viruses [12], three beet poleroviruses in sugarbeet and aphids [38, 39], eight wheat viruses [13], nine crinivirus infecting vegetable and small fruit crops [15], four viruses infecting cassava [18], five tobacco viruses [19], three cucurbit-infecting poleroviruses [30], seven main tomato-infecting viruses [20], three rice viruses [21], three lily-infecting viruses [22], four cherry viruses [23], four viruses in peach [24], three viruses in pear plants [40], five viruses and two viriods infecting chrysanthemum [41], three viruses infecting papaya [42], three orchid viruses [25], five fabaviruses [31] and so on.…”

Section: Discussionmentioning

confidence: 99%

Simultaneous detection and differentiation of three genotypes of Brassica yellows virus by multiplex reverse transcription-polymerase chain reaction

Zhang

Peng

Wang

et al. 2016

Virol J

View full text Add to dashboard Cite

BackgroundBrassica yellows virus (BrYV), proposed to be a new polerovirus species, three distinct genotypes (BrYV-A, BrYV-B and BrYV-C) have been described. This study was to develop a simple, rapid, sensitive, cost-effective method for simultaneous detection and differentiation of three genotypes of BrYV.ResultsIn this study, a multiplex reverse transcription-polymerase chain reaction (mRT-PCR) was developed for simultaneous detection and differentiation of the three genotypes of BrYV. The three genotypes of BrYV and Tunip yellows virus (TuYV) could be differentiated simultaneously using six optimized specific oligonucleotide primers, including one universal primer for detecting BrYV, three BrYV genotype-specific primers, and a pair of primers for specific detection of TuYV. Primers were designed from conserved regions of each virus and their specificity was confirmed by sequencing PCR products. The mRT-PCR products were 278 bp for BrYV-A, 674 bp for BrYV-B, 505 bp for BrYV-C, and 205 bp for TuYV. Amplification of three target genotypes was optimized by increasing the PCR annealing temperatures to 62 °C. One to three fragments specific for the virus genotypes were simultaneously amplified from infected samples and identified by their specific molecular sizes in agarose gel electrophoresis. No specific products could be amplified from cDNAs of other viruses which could infect crucifer crops. Detection limits of the plasmids for multiplex PCR were 100 fg for BrYV-A and BrYV-B, 10 pg for BrYV-C, and 1 pg for TuYV, respectively. The mRT-PCR was applied successfully for detection of three BrYV genotypes from field samples collected in China.ConclusionsThe simple, rapid, sensitive, and cost-effective mRT-PCR was developed successfully for detection and differentiation of the three genotypes of BrYV.

show abstract

Section: Discussionmentioning

confidence: 99%

Simultaneous detection and differentiation of three genotypes of Brassica yellows virus by multiplex reverse transcription-polymerase chain reaction

Zhang

Peng

Wang

et al. 2016

Virol J

View full text Add to dashboard Cite

show abstract

“…In this report, we describe a method of two-step multiplex RT-PCR assay for detection of four stone fruit virus species. So far, to our knowledge, several reports about stone fruit virus detection have been published (Sanchez-Navarro et al, 2005;Jarosova and Kundu, 2010;Noorani et al, 2013;Yu et al, 2013). However, owing to the different virus combinations it chose in the test, most of the developed methods cannot be used directly in our surveys.…”

Section: Discussionmentioning

confidence: 99%

“…However, owing to the different virus combinations it chose in the test, most of the developed methods cannot be used directly in our surveys. Taking Noorani's research for example, Cherry virus A (CVA), Little cherry virus-1 (LChV-1), Cherry necrotic rusty mottle virus (CNRMV) and PNRSV were detected simultaneously in their assay (Noorani et al, 2013). However, LChV-1 has never been identified from stone fruit trees in China.…”

Section: Discussionmentioning

confidence: 99%

See 1 more Smart Citation

A multiplex RT-PCR assay for simultaneous detection of four viruses from sweet cherry

Zong

Wang

Wei

et al. 2014

Scientia Horticulturae

View full text Add to dashboard Cite

“…Multiplex reverse transcriptase PCR (mRT‐PCR) has been used to simultaneously amplify more than one locus in the same reaction, proving to be a rapid, cost‐effective screening assay for plant viruses (Bariana et al ; Nemchinov et al ; Henegariu et al ; Matic et al ; Noorani et al ; Ge et al ). Various types of mRT‐PCR assays have been previously applied to detect viruses that affect stone fruit trees.…”

Section: Introductionmentioning

confidence: 99%

Multiplex RT‐PCR to simultaneously detect three viruses that infect peach

Yang

Zhou

et al. 2019

Lett Appl Microbiol

View full text Add to dashboard Cite

Significance and Impact of the Study: In this study, we developed a multiplex reverse transcriptase PCR (mRT-PCR) assay for simultaneously detecting three viruses that infect peach: peach-associated luteovirus (PaLV), peach virus D (PeVD) and nectarine stem-pitting-associated virus (NSPaV). This assay is simple, easy to perform, reliable and cost-effective, and can thus be applied for large-scale surveys of PaLV, PeVD and NSPaV. AbstractPeach is a major crop in China, and like any other stone fruit, virus and viruslike diseases can reduce the yield and quality of the fruit. Herein, we developed a multiplex RT-PCR (mRT-PCR) assay for simultaneously detecting three viruses known to infect peach: peach-associated luteovirus (PaLV), peach virus D (PeVD) and nectarine stem-pitting-associated virus (NSPaV). Plant nad5 mRNA was used as the internal control. Field samples that were co-infected with PaLV, PeVD and NSPaV were used; we identified three primer pairs to be the most specific for detecting these viruses, followed by determining the ideal concentration of each primer pair and optimizing the annealing temperature for mRT-PCR. We also assessed the detection limit using serial dilutions of RNA and cDNA. The newly developed mRT-PCR assay could simultaneously detect PaLV, PeVD and NSPaV. To validate the reliability of mRT-PCR for virus detection, mRT-PCR was used to detect viruses in the leaves of 21 peach plants collected in Liaoning Province, China. The obtained results revealed the presence of single and co-infections. To conclude, the mRT-PCR assay developed herein is sensitive, reliable and economical, and we believe that it can thus be used for large-scale surveys of PaLV, PeVD and NSPaV.Letters in Applied Microbiology ISSN 0266-8254

show abstract

Simultaneous detection and identification of four cherry viruses by two step multiplex RT-PCR with an internal control of plant nad5 mRNA

Cited by 26 publications

References 38 publications

Simultaneous detection and differentiation of three genotypes of Brassica yellows virus by multiplex reverse transcription-polymerase chain reaction

Simultaneous detection and differentiation of three genotypes of Brassica yellows virus by multiplex reverse transcription-polymerase chain reaction

A multiplex RT-PCR assay for simultaneous detection of four viruses from sweet cherry

Multiplex RT‐PCR to simultaneously detect three viruses that infect peach

Contact Info

Product

Resources

About