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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 #433822

Research Project: An NGS-based System for Unambiguous Detection of HLB Pathogen

Location: Crop Diseases, Pests and Genetics Research

Project Number: 2034-22000-012-14-R
Project Type: Reimbursable Cooperative Agreement

Start Date: Oct 1, 2017
End Date: Sep 30, 2019

Objective:
The aim is to develop a detection system that can unambiguously identify "Candidatus Liberibacter asiaticus", an unculturable bacterium associated with Huanglongbing (HLB), when the bacterial concentration is very low and current polymerase chain reaction (PCR) techniques could not make a clear conclusion. There are three objectives: 1. Effective enrichment of low titer CLas DNA from HLB suspected sample; 2. Identify a cost-effective protocol for sample sequencing; and 3. Evaluate and finalize an analytical pipeline for accurate and sensitive detection of CLas.

Approach:
The system will make direct identification of sequences of PCR template using the up-to-date next generation sequencing/high throughput sequencing technology. Sequence data will be compared to signature sequences of "Candidatus Liberibacter asiaticus" of taxonomic authority. The detection result will be either a yes (matched) or a no (un-matched), without ambiguity.