Development of a Liquichip-Based Assay for Detection of Paracidovorax citrulli in Cucurbits
Other Title
建立以Liquichip為基礎之葫蘆科作物中Paracidovorax citrulli檢測分析方法
Journal
台灣農業研究
Journal Volume
75
Journal Issue
2
Start Page
281
End Page
291
ISSN
2790-086X
Date Issued
2026-06-08
Author(s)
Abstract
Bacterial fruit blotch (BFB), caused by Paracidovorax citrulli, threatens cucurbit production worldwide and often results in severe economic losses. In this study, we developed a detection assay for sensitive detection of P. citrulli. The assays employ polymerase chain reaction (PCR) with specific co-amplification of a plant gene as an internal control from total nucleic acids, enabling parallel detection and minimizing the risk of false-negative PCR results during routine testing and eliminating the need to remove contaminating plant DNA from extracts. The assay exhibited high specificity, as no cross-reactions were observed with non-target bacterial species or with nucleic acid extracted from healthy cucurbit tissues. Sensitivity testing demonstrated reliable detection of low concentrations of pathogen DNA, and the method was validated using artificially inoculated seedlings and field samples. Compared with conventional PCR, the multiplex detection assay provides higher throughput and quantitative readouts based on mean fluorescence intensity. This platform offers a reliable tool for seed health testing, quarantine surveillance, and integrated management of BFB in cucurbit crops.
由 Paracidovorax citrulli 引起的細菌性果斑病 (bacterial fruit blotch; BFB) 對全球瓜類作物生產造成嚴重威脅，並經常導致顯著的經濟損失。本研究建立一套可高靈敏偵測 P. citrulli 的檢測方法。該檢測系統採用聚合酶連鎖反應 (polymerase chain reaction; PCR) 並同時共擴增植物基因作為內部控制，以全核酸進行分析偵測，有效降低例行檢測中產生偽陰性結果的風險，並降低樣品中植物 DNA 污染。結果顯示，本方法具有高度專一性，未與非目標細菌菌種或健康瓜類組織所萃取之 DNA 產生交叉反應。靈敏度測試證實本系統可穩定偵測低濃度之病原菌 DNA，且已成功應用於人工接種幼苗感染田間樣品之驗證。與傳統 PCR 方法相比，本檢測系統具備較高的檢測通量，並可依螢光強度提供定量化的分析結果。本平臺可作為瓜類果斑病於種子健康檢測、檢疫監測及整合性病害管理之可靠診斷工具。
Subjects
Paracidovorax citrulli
Bacterial fruit blotch
Cucurbits
Diagnosis
細菌性果斑病
瓜類
分子診斷
西瓜
Publisher
農業部農業試驗所
Type
journal article
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