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

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. [ABSTRACT FROM AUTHOR]