Detection of Tobacco Ringspot Virus in Soybean Seeds by the Reverse Transcription Loop-mediated Isothermal Amplification Method(PDF)

《大豆科学》[ISSN:1000-9841/CN:23-1227/S]

Issue:

2012年06期

Page:

980-983,987

Research Field:

Publishing date:

Info

Title:

Detection of Tobacco Ringspot Virus in Soybean Seeds by the Reverse Transcription Loop-mediated Isothermal Amplification Method

Author(s):

GUO Li-xin¹; CHEN Xian-feng²; XU Ya-fei¹; QIU Zhi-jun¹; ZHU Di-qi¹; DUAN Wei-jun²

1.Beilun Entry-exit Inspection and Quarantine Bureau,Ningbo 315800,Zhejiang;?

2.Ningbo Entry-exit Inspection and Quarantine Bureau,Ningbo 315012,Zhejiang,China

Keywords:

Tobacco ringspot virus(TRSV); Reverse transcription loop-mediated isothermal amplification(RT-LAMP); Detection

PACS:

S412

DOI:

10.3969/j.issn.1000-9841.2012.06.026

Abstract:

A sets of primers were designed and synthesized based on the coat protein coding region of tobacco ringspot virus(TRSV)for reverse transcription loop-mediated isothermal amplification(RT-LAMP)assay.The amplification was detected using the real time turbidimeter.An accelerated RT-LAMP procedure with one step was developed for the detection of TRSV.The specificity and sensitivity of the RT-LAMP assay were tested.RT-PCR compared to RT-LAMP by serial dilutions of total RNA extracts,they had the same sensitivities,but RT-LAMP was quicker,and the reaction could be finished within 1 hour.In addition,the presence or absence of the fluorescent display in daylight allowed naked easy detection of the amplification of TRSV genomic RNA using Calcein.The RT-LAMP assay was applied successfully to detect TRSV in soybean seeds,and the result by the addition of Calcein was consistent with the result detected by the real time turbidimeter.The method is rapid,specific,sensitive and suitable for rapid field detection of TRSV.

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