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Journal of Integrative Agriculture  2018, Vol. 17 Issue (2): 483-487    DOI: 10.1016/S2095-3119(17)61815-X
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Development of a sensitive and reliable droplet digital PCR assay for the detection of ‘Candidatus Liberibacter asiaticus’
ZHONG Xi1, LIU Xue-lu1, LOU Bing-hai2, ZHOU Chang-yong1, WANG Xue-feng
1 National Citrus Engineering Research Center, Citrus Research Institute, Chinese Academy of Agricultural Sciences/Southwest University, Chongqing 400712, P.R.China
2 Guangxi Key Laboratory of Citrus Biology, Guangxi Academy of Specialty Crops, Guilin 541004, P.R.China
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Abstract  Citrus Huanglongbing (HLB, yellow shoot disease) is one of the most serious citrus diseases worldwide.  To better improve the detection sensitivity, a droplet digital PCR (ddPCR) assay was developed for the rapid detection of ‘Candidatus Liberibacter asiaticus’ (Las), the putative causal agent of HLB.  The detection of sensitivity comparison using positive plasmid indicated that ddPCR was superior to quantitative PCR (qPCR) for detecting and quantifying Las at low concentrations.  The Las detection of 40 field samples also showed that six of 13 asymptomatic samples (46.15%) with high Ct value (>35) were positive by ddPCR.  This methodology showed great potential for early HLB infection diagnosis.
Keywords:  citrus Huanglongbing        early diagnosis        droplet digital PCR  
Received: 10 July 2017   Accepted:
Fund: 

This study was funded by the National Natural Sciences Foundation of China (31671992, 31301635), the Chongqing Science and Technology Commission Project, China (cstc2016shms-ztzx80003) and the Guangxi Key Laboratory of Citrus Biology, Guangxi Academy of Specialty Crops, China (SYS2015K004).

Corresponding Authors:  Correspongdence WANG Xue-feng, Tel: +86-23-68349338; E-mail: wangxuefeng@cric.cn   
About author:  ZHONG Xi, E-mail:zhongmxi@yeah.net

Cite this article: 

ZHONG Xi, LIU Xue-lu, LOU Bing-hai, ZHOU Chang-yong, WANG Xue-feng . 2018. Development of a sensitive and reliable droplet digital PCR assay for the detection of ‘Candidatus Liberibacter asiaticus’. Journal of Integrative Agriculture, 17(2): 483-487.

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