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Simultaneous detection and differentiation of four closely related sweet potato potyviruses by a multiplex one-step RT-PCR. 2012

Fan Li, and Ruijuan Zuo, and Jorge Abad, and Donglin Xu, and Gaili Bao, and Ruhui Li
USDA-ARS, National Germplasm Resources Laboratory, Beltsville, MD 20705, USA.

Four closely related potyviruses, Sweet potato feathery mottle virus (SPFMV), Sweet potato virus C (SPVC), Sweet potato virus G (SPVG) and/or Sweet potato virus 2 (SPV2), are involved in sweet potato virus disease complexes worldwide. Identification and detection of these viruses are complicated by high similarity among their genomic sequences, frequent occurrence as mixed infections and low titer in many sweet potato cultivars. A one-tube multiplex reverse transcription-PCR (mRT-PCR) assay was developed for simultaneous detection and differentiation of SPFMV, SPVC, SPVG and SPV2. Four specific forward primers unique to each virus and one reverse primer based on the region conserved in all four viruses were selected and used in the assay. The mRT-PCR assay was optimized for primer concentration and cycling conditions. It was tested using sweet potato plants infected naturally with one to four target viruses and then evaluated using field samples collected from southwestern China. The mRT-PCR assay is reliable and sensitive as a simple, rapid and cost-effective method to detect these pathogens in sweet potato. This assay will be useful to quarantine and certification programs and virus surveys when large numbers of samples are tested.

UI MeSH Term Description Entries
D010935 Plant Diseases Diseases of plants. Disease, Plant,Diseases, Plant,Plant Disease
D002681 China A country spanning from central Asia to the Pacific Ocean. Inner Mongolia,Manchuria,People's Republic of China,Sinkiang,Mainland China
D012680 Sensitivity and Specificity Binary classification measures to assess test results. Sensitivity or recall rate is the proportion of true positives. Specificity is the probability of correctly determining the absence of a condition. (From Last, Dictionary of Epidemiology, 2d ed) Specificity,Sensitivity,Specificity and Sensitivity
D014773 Virology The study of the structure, growth, function, genetics, and reproduction of viruses, and VIRUS DISEASES.
D060885 Multiplex Polymerase Chain Reaction Methods for using more than one primer set in a polymerase chain reaction to amplify more than one segment of the target DNA sequence in a single reaction. Multiplex Ligation-Dependent Probe Amplification,Multiplex PCR,Triplex PCR,Triplex Polymerase Chain Reaction,Multiplex Ligation Dependent Probe Amplification,PCR, Multiplex,PCR, Triplex
D017800 Potyvirus A large genus of plant viruses of the family POTYVIRIDAE which infect mainly plants of the Solanaceae. Transmission is primarily by aphids in a non-persistent manner. The type species is potato virus Y. Potato Virus Y,Potato Virus Ys,Potyviruses,Virus Ys, Potato,Ys, Potato Virus
D017931 DNA Primers Short sequences (generally about 10 base pairs) of DNA that are complementary to sequences of messenger RNA and allow reverse transcriptases to start copying the adjacent sequences of mRNA. Primers are used extensively in genetic and molecular biology techniques. DNA Primer,Oligodeoxyribonucleotide Primer,Oligodeoxyribonucleotide Primers,Oligonucleotide Primer,Oligonucleotide Primers,Primer, DNA,Primer, Oligodeoxyribonucleotide,Primer, Oligonucleotide,Primers, DNA,Primers, Oligodeoxyribonucleotide,Primers, Oligonucleotide
D020133 Reverse Transcriptase Polymerase Chain Reaction A variation of the PCR technique in which cDNA is made from RNA via reverse transcription. The resultant cDNA is then amplified using standard PCR protocols. Polymerase Chain Reaction, Reverse Transcriptase,Reverse Transcriptase PCR,PCR, Reverse Transcriptase,Transcriptase PCR, Reverse
D027723 Ipomoea batatas A plant species of the genus IPOMOEA, family CONVOLVULACEAE. Some cultivars are sweet and edible whereas bitter varieties are a source of SAPONINS. This sweet potato is sometimes referred to as a yam (DIOSCOREA). Sweet Potato,Yam, Pseudo,Pseudo-Yam,Ipomoea batata,Potato, Sweet,Potatos, Sweet,Pseudo Yam,Pseudo Yams,Pseudo-Yams,Sweet Potatos,Yams, Pseudo,batatas, Ipomoea

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