A sensitive semi-quantitative biotin-streptavidin ELISA for the detection of potyviruses infecting cucurbits

Dietzgen, R. G.; Herrington, M. E.

doi:10.1071/ar9910417

Cited by 9 publications

(5 citation statements)

References 22 publications

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“…Menassa et al (1986) described the detection of ZYMV in intact leaf disks by direct or indirect ELISA tests; attempts to detect ZYMV in viruliferous aphids were not satisfactory. Dietzgen & Herrington (1991) used a semiquantitative biotinstreptavidin ELISA, with a sensitivity increased 4-8 times compared to DAS-ELISA.…”

Section: Serological Techniquesmentioning

confidence: 99%

Zucchini yellow mosaic virus

Desbiez¹,

Lecoq²

1997

Plant Pathology

224

172

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Zucchini yellow mosaic potyvirus (ZYMV), first isolated in Italy in 1973, described in 1981, and then identified in all continents within a decade, is one of the most economically important viruses of cucurbit crops. It is efficiently aphid-transmitted in a nonpersistent manner and it is also seed-borne in zucchini squash, which could have contributed to its rapid spread worldwide. Biological variability has been observed among ZYMV isolates, concerning host range, symptomatology and aphid transmissibility. More recent studies also revealed a serological and molecular variability. The survival of ZYMV in areas where cucurbits are not grown throughout the year remains to be elucidated, because very few natural over-wintering hosts have been identified so far. Partial control of ZYMV can be achieved by limiting transmission of the virus to the crops by aphids, using adapted cultural practices. Cross-protection with a mild strain has been shown to be effective against most ZYMV isolates. Resistance genes found in cucurbit germplasms are currently being introduced into cultivars with good agronomical characteristics. Pathogen-derived resistance strategies using the expression of ZYMV genes in transgenic plants have also been developed and appear promising. Nevertheless, the high biological variability of ZYMV justifies a careful evaluation of the deployment of genetic control strategies in order to increase their durability.

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Section: Serological Techniquesmentioning

confidence: 99%

Zucchini yellow mosaic virus

Desbiez¹,

Lecoq²

1997

Plant Pathology

224

172

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“…Other diagnostic methods reported for detection of ZYMV are immunobinding, ELISA, RT‐PCR/PCR and real‐time PCR (Somowiyarjo et al. ; Dietzgen and Herrington ; Thomson et al. ; Coutts et al.…”

Section: Discussionmentioning

confidence: 99%

“…Dietzgen and Herrington () reported that RT‐PCR is more effective and more specific than ELISA for the detection of ZYMV in diseased cucurbitaceous plants. However, our study indicates that the RT‐LAMP assay is superior to the RT‐PCR assay in detection of ZYMV in infected plant tissues because the test is faster than the conventional RT‐PCR.…”

Section: Discussionmentioning

confidence: 99%

Use of Reverse Transcription Loop‐Mediated Isothermal Amplification for the Detection of Zucchini yellow mosaic virus

Kuan

Deng

Huang

et al. 2013

Journal of Phytopathology

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A Reverse Transcription Loop-Mediated Isothermal Amplification (RT-LAMP) assay was employed to develop a simple and efficient system for the detection of Zucchini yellow mosaic virus (ZYMV) in squash and melon plants. The RT-LAMP assay took 30 min under isothermal condition at 64°C by employing a set of four primers targeting ZYMV. The sensitivity of RT-LAMP was 10-fold greater than that of the RT-PCR assay in the detection of ZYMV in infected tissues of squash and melon. No reaction was detected from the tissues of healthy plants by either RT-LAMP or RT-PCR assay. The RT-LAMP product of the tested samples can be visualized by staining directly in the tube with SYBR Green I dye. The sensitivity of SYBR Green I staining method is similar to that analyzed by gel electrophoresis. Field-grown squash and melon plants were tested using RT-PCR and RT-LAMP. Both RT-LAMP and PCR could detect ZYMV in symptomatic or symptomless tissues of infected plants. However, the RT-LAMP assay is superior to RT-PCR because it is rapid, simple, and highly sensitive; therefore, RT-LAMP is a useful and practical method for detection of ZYMV in cucurbits.

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“…Both serological and molecular diagnostic methods are available for the detection of WMV and ZYMV in routine diagnosis, such as enzyme-linked immunosorbent analysis (ELISA) [ 17 ], conventional reverse-transcription polymerase chain reaction (RT-PCR) [ 6 , 18 , 19 , 20 ], reverse transcription loop-mediated isothermal amplification (RT-LAMP) [ 21 , 22 ], and small RNA sequencing [ 11 ]. Most of these methods were not validated for their sensitivity and specificity according to the international requirements for plant pest diagnosis.…”

Section: Introductionmentioning

confidence: 99%

Development, Validation, and Application of Reverse Transcription Real-Time and Droplet Digital PCR Assays for the Detection of the Potyviruses Watermelon Mosaic Virus and Zucchini Yellow Mosaic Virus in Cucurbits

Luigi

Manglli

Corrado

et al. 2023

Plants

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Among the cucurbit-infecting viruses, watermelon mosaic virus (WMV) and zucchini yellow mosaic virus (ZYMV) (Potyvirus: Potyviridae) are responsible for severe symptoms on cucumber, melon, watermelon, and zucchini cultivations worldwide. In this study, reverse transcription real-time PCR (real-time RT-PCR) and droplet-digital PCR (RT-ddPCR) assays targeting the coat protein (CP) genes of WMV and ZYMV were developed and validated according to the international standards of plant pest diagnosis (EPPO PM 7/98 (5)). First, the diagnostic performance of WMV-CP and ZYMV-CP real-time RT-PCRs was evaluated, and the assays displayed an analytical sensitivity of 10−5 and 10−3, respectively. The tests also showed an optimal repeatability, reproducibility and analytical specificity, and were reliable for the virus detection in naturally infected samples and across a wide range of cucurbit hosts. Based on these results, the real-time RT-PCR reactions were adapted to set up RT-ddPCR assays. These were the first RT-ddPCR assays aiming at the detection and quantification of WMV and ZYMV and showed a high sensitivity, being able to detect until 9 and 8 copies/µL of WMV or ZYMV, respectively. The RT-ddPCRs allowed the direct estimation of the virus concentrations and opened to a broad range of applications in disease management, such as the evaluation of partial resistance in breeding processes, identification of antagonistic/synergistic events, and studies on the implementation of natural compounds in the integrated management strategies.

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A sensitive semi-quantitative biotin-streptavidin ELISA for the detection of potyviruses infecting cucurbits

Cited by 9 publications

References 22 publications

Zucchini yellow mosaic virus

Zucchini yellow mosaic virus

Use of Reverse Transcription Loop‐Mediated Isothermal Amplification for the Detection of Zucchini yellow mosaic virus

Development, Validation, and Application of Reverse Transcription Real-Time and Droplet Digital PCR Assays for the Detection of the Potyviruses Watermelon Mosaic Virus and Zucchini Yellow Mosaic Virus in Cucurbits

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