Development of a one-step immunocapture real-time TaqMan RT-PCR assay for the broad spectrum detection of Pepino mosaic virus.

Development of a one-step immunocapture real-time TaqMan RT-PCR assay for the broad spectrum detection of Pepino mosaic virus. Research Abstract Details 

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Development of a one-step immunocapture real-time TaqMan RT-PCR assay for the broad spectrum detection of Pepino mosaic virus. Abstract Text:

Kai-Shu Ling,W Patrick Wechter,Ramon Jordan,

A real-time reverse-transcription polymerase chain reaction (RT-PCR) was developed for efficient detection of genetically diverse Pepino mosaic virus (PepMV) isolates. The novel detection system was designed to use a duo-primer system targeting the conserved region in the triple gene block 2 (TGB2) gene with a single conserved TaqMan probe to broaden its reaction to cover all available PepMV strains. This duo-primer real-time RT-PCR assay was evaluated against US1, US2, Ch1, Ch2 and 25 field isolates collected from six major commercial tomato greenhouse facilities in U.S. and Canada in 2006. Under optimum reaction conditions, sensitivity of the detection was as low as 100 fg of purified viral RNA. This assay was also evaluated for its efficiency in detecting PepMV in various levels of contaminated seed samples. Using immuno-capture sample preparation, real-time RT-PCR was able to detect PepMV in one infested seed in 1000. This level of sensitivity indicated that the one-step immuno-capture duo-primer TaqMan real-time RT-PCR developed in the present study could be used for routine seed health assays.

Development of a one-step immunocapture real-time TaqMan RT-PCR assay for the broad spectrum detection of Pepino mosaic virus. Publishing Authors By Initials

KS Ling,WP Wechter,R Jordan,

For similar enzymes and coenzymes: enzymes: transferases: phosphotransferases: nucleotidyltransferases: dna nucleotidyltransferases: dna-directed dna polymerase: taq polymerase research abstracts see: enzymes and coenzymes: enzymes: transferases: phosphotransferases: nucleotidyltransferases: dna nucleotidyltransferases: dna-directed dna polymerase: taq polymerase research

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Development of a one-step immunocapture real-time TaqMan RT-PCR assay for the broad spectrum detection of Pepino mosaic virus. Journal Published:

PUBLICATION TYPE: Journal Article

Journal: Journal of virological methods

VOLUME: 144

Page Numbers: 65-72

Journal Abbreviation: J. Virol. Methods

ISSN: 0166-0934

DAY: 11

MONTH: 05

YEAR: 2007

Development of a one-step immunocapture real-time TaqMan RT-PCR assay for the broad spectrum detection of Pepino mosaic virus. Information

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LANGUAGE: eng

NlmUniqueID: 8005839

Development of a one-step immunocapture real-time TaqMan RT-PCR assay for the broad spectrum detection of Pepino mosaic virus. Keywords Mesh Terms:

KEYWORDS: Taq Polymerase

MESH TERMS: genetics

Chemical & Substance for Abstract: Development of a one-step immunocapture real-time TaqMan RT-PCR assay for the broad spectrum detection of Pepino mosaic virus. Information

Substance Name: Taq Polymerase

Registry Number: EC 2.7.7.-

Grant and Affiliation Information for Development of a one-step immunocapture real-time TaqMan RT-PCR assay for the broad spectrum detection of Pepino mosaic virus.

AFFILIATION: USDA-ARS, U.S. Vegetable Laboratory, 2700 Savannah Highway, Charleston, SC, USA. kai.ling@ars.usda.gov

Country: Netherlands

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MEDLINETA: J Virol Methods

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