A more sensitive and rapid multiplex RT-PCR assay combining with magnetic nanobeads for simultaneous detection of viruses in sweet potato

Deng, Xing-Guang; Zhu, Feng; Chen, Yingjuan; Liu, Jian; Zhu, Tong; Li, Jingyi; Xi, Dehui; Lin, Honghui

doi:10.1007/s10658-014-0447-0

Cited by 6 publications

(2 citation statements)

References 32 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Some of these limitations have been overcome by improving the quality of the nucleic acid extraction procedures, the products obtained and/or different modalities of the PCR technology. Magnetic nanobeads ( Deng et al, 2014 ), dual priming oligonucleotide (DPO) primers ( Kwon et al, 2014 ), or a nested reaction ( Foissac et al, 2005 ; Maliogka et al, 2007 ; Papayiannis et al, 2011 ) have all contributed to increased levels of specificity and sensitivity.…”

Section: Multiplex Pcrmentioning

confidence: 99%

Recent Advances on the Multiplex Molecular Detection of Plant Viruses and Viroids

2018

View full text Add to dashboard Cite

Plant viruses are still one of the main contributors to economic losses in agriculture. It has been estimated that plant viruses can cause as much as 50 billion euros loss worldwide, per year. This situation may be worsened by recent climate change events and the associated changes in disease epidemiology. Reliable and early detection methods are still one of the main and most effective actions to develop control strategies for plant viral diseases. During the last years, considerable progress has been made to develop tools with high specificity and low detection limits for use in the detection of these plant pathogens. Time and cost reductions have been some of the main objectives pursued during the last few years as these increase their feasibility for routine use. Among other strategies, these objectives can be achieved by the simultaneous detection and (or) identification of several viruses in a single assay. Nucleic acid-based detection techniques are especially suitable for this purpose. Polyvalent detection has allowed the detection of multiple plant viruses at the genus level. Multiplexing RT polymerase chain reaction (PCR) has been optimized for the simultaneous detection of more than 10 plant viruses/viroids. In this short review, we provide an update on the progress made during the last decade on techniques such as multiplex PCR, polyvalent PCR, non-isotopic molecular hybridization techniques, real-time PCR, and array technologies to allow simultaneous detection of multiple plant viruses. Also, the potential and benefits of the powerful new technique of deep sequencing/next-generation sequencing are described.

show abstract

Section: Multiplex Pcrmentioning

confidence: 99%

Recent Advances on the Multiplex Molecular Detection of Plant Viruses and Viroids

2018

View full text Add to dashboard Cite

show abstract

“…In multiplex RT-PCR system, chemical components such as Taq polymerase and dNTPs, are competitively consumed to simultaneously amplify multiple sequences in one reaction. It may be the main reason for the difference in detection sensitivity between multiplex RT-PCR and monoplex RT-PCR.Previous studies showed that the decreased sensitivity, commonly 1-2 orders of magnitude in comparison to monoplex RT-PCR, was not a major limiting factor in using multiplex RT-PCR for virus detectionDeng et al, 2014).…”

mentioning

confidence: 99%

A developed multiplex RT-PCR assay for simultaneous detection of four main viruses on pepper (Capsicum spp.)

Gong,

Zhao,

et al. 2023

Preprint

View full text Add to dashboard Cite

Viral diseases cause severe losses commonly and become one of main global limiting factors in pepper production. Chilli veinal mottle virus (ChiVMV), pepper veinal mottle virus (PVMV), cucumber mosaic virus (CMV), and pepper mild mottle virus (PMMoV) are the dominant viruses damaging pepper in South China. To detect these viruses efficiently, a developed multiplex reverse transcription polymerase chain reaction (RT-PCR) method was established. Four pairs of specific primers were used to amplify a 990 bp product for PMMoV, a 923 bp product for ChiVMV, a 823 bp product for PVMV, and a 682 bp product for CMV. The optimal primer concentration, the optimal reaction annealing temperature and cycle number were determined. The developed multiplex RT-PCR could detect PMMoV, ChiVMV, CMV and PVMV from cDNA diluted up to 10–4, 10–4, 10–3 and 10–3, respectively. Then it was sucessfully used to detect virus infection from eight mixed pepper leaf samples. The result were basically consistent with that of monoplex RT-PCR. Additionally, seventy-five diseased samples collected from nine major pepper growing regions in Guangxi were analyzed by the developed multiplex RT-PCR. The results showed that most of samples were coinfected by two or more viruses, and ChiVMV and PVMV had relatively higher total detection rates of 65.33% and 60.00% respectivlely. The results also indicated that the four viruses infecting pepper weren't evenly distributed in Guangxi. In a conclusion, the developed multiplex RT-PCR here will be a efficient tool for routine molecular diagnosis of PMMoV, ChiVMV, PVMV and CMV.

show abstract

A multiple reverse transcription PCR assay for simultaneous detection of four main viruses in kiwifruit

et al. 2020

View full text Add to dashboard Cite

A more sensitive and rapid multiplex RT-PCR assay combining with magnetic nanobeads for simultaneous detection of viruses in sweet potato

Cited by 6 publications

References 32 publications

Recent Advances on the Multiplex Molecular Detection of Plant Viruses and Viroids

Recent Advances on the Multiplex Molecular Detection of Plant Viruses and Viroids

A developed multiplex RT-PCR assay for simultaneous detection of four main viruses on pepper (Capsicum spp.)

A multiple reverse transcription PCR assay for simultaneous detection of four main viruses in kiwifruit

Contact Info

Product

Resources

About