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ARS Home » Pacific West Area » Parlier, California » San Joaquin Valley Agricultural Sciences Center » Crop Diseases, Pests and Genetics Research » Research » Research Project #434468

Research Project: Improved Sensitive Detection of the HLB Pathogen by New Gene Primers for Real Time and Droplet Digital PCR Analyses

Location: Crop Diseases, Pests and Genetics Research

Project Number: 2034-22000-013-12-T
Project Type: Trust Fund Cooperative Agreement

Start Date: Jan 1, 2018
End Date: Dec 31, 2018

Objective:
The objective is to improve reliable diagnosis of "Candidatus Liberibacter asiaticus", presumptive bacterial causal agent of huanglongbing (HLB) (aka greening) and to more readily distinguish between HLB and Spiroplasma citri, the bacterial causal agent of citrus stubborn disease.

Approach:
Deep sequencing using MiSeq PacBio platform to obtain additional full-length seqeunces of "Candidatus Liberibacter asiaticus" and Spiroplasma citri; conduct bioinformatic analysis, design new primers and probe (Taqman) with appropriate software and develop a new isothermal amplification assay using loop-mediated (LAMP) or Recombinase Polymerase Amplification (RPA) along with real time quantitative PCR, droplet digital PCR, high throughput robotic DNA extraction and purification of nucleic acids.