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United States Department of Agriculture

Agricultural Research Service

Research Project: GENETIC AND BIOLOGICALLY-BASED MANAGEMENT OF VEGETABLE CROP DISEASES

Location: Vegetable Research

Title: Sensitive and Cost-Effective Immunocapture RT-PCR for Routinely Viral Detection in Large Number of Plant Samples

Authors
item Xia, Jun -
item Ling, Kai-Shu

Submitted to: Phytopathology
Publication Type: Abstract Only
Publication Acceptance Date: May 1, 2010
Publication Date: June 1, 2010
Citation: Xia, J.Q., Ling, K. 2010. Sensitive and Cost-Effective Immunocapture RT-PCR for Routinely Viral Detection in Large Number of Plant Samples. Phytopathology. 100-S139.

Technical Abstract: It has long been a challenging task to develop timely and accurate diagnostic tests for diseases caused by viruses or virus-like agents in plants in agriculture production. Current ELISA detection methods may not provide the sensitivity needed for samples with low viral concentrations. Conventional RT-PCR, which requires relatively pure nucleic acid samples, is costly and time-consuming. By combining two widely used virus detection methods, ELISA and RT-PCR, Immunocapture RT-PCR was developed for practical detection of plant viruses. The immunocapture sample preparation allows RT-PCR to be performed in a 96-well format equivalent to that of a regular ELISA test. The entire RT-PCR assay, viral immunocapturing from samples, captured viral RNA amplification and amplicon detection, can be conducted in a single PCR reaction tube within a relatively short time. This novel technology makes it possible to routinely detect plant viruses in large number of samples by RT-PCR. This sensitive and cost-effective immunocapture RT-PCR assay has huge potential applications in plant viral disease diagnosis.

Last Modified: 8/29/2014
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