A CRISPR/Cas12a-based portable platform for rapid detection of Leptosphaeria maculans in Brassica crops

作者全名："Lei, Rong; Li, Yuan; Li, Limei; Wang, Jingyi; Cui, Zhenhai; Ju, Rui; Jiang, Li; Liao, Xiaoling; Wu, Pinshan; Wang, Xinyi"

作者地址："[Lei, Rong; Jiang, Li; Wu, Pinshan] Chinese Acad Inspect & Quarantine, Beijing, Peoples R China; [Li, Yuan] Chongqing Med Univ, Yongchuan Hosp, Cent Lab, Chongqing, Peoples R China; [Li, Limei; Wang, Xinyi] Dalian Univ, Sch Life & Hlth, Dalian, Peoples R China; [Li, Limei; Wang, Jingyi; Ju, Rui] Shenyang Agr Univ, Coll Sci, Shenyang, Peoples R China; [Cui, Zhenhai] Chinese Acad Sci, Northeast Inst Geog & Agroecol, Key Lab Soybean Mol Design & Breeding, Changchun, Peoples R China; [Liao, Xiaoling] Chongqing Univ Sci & Technol, Coll Met & Mat Engn, Chongqing, Peoples R China"

通信作者："Wu, PS (通讯作者)，Chinese Acad Inspect & Quarantine, Beijing, Peoples R China.; Wang, XY (通讯作者)，Dalian Univ, Sch Life & Hlth, Dalian, Peoples R China.; Liao, XL (通讯作者)，Chongqing Univ Sci & Technol, Coll Met & Mat Engn, Chongqing, Peoples R China."

来源：FRONTIERS IN PLANT SCIENCE

ESI学科分类：PLANT & ANIMAL SCIENCE

WOS号：WOS:000859842300001

JCR分区：Q1

影响因子：5.6

年份：2022

卷号：13

期号：　

开始页：　

结束页：　

文献类型：Article

关键词：L; maculans; recombinase polymerase amplification (RPA); Cas12a; all-in-one chip; lateral flow assay; portable platform

摘要："Establishing a portable diagnostic method for identifying plant pathogens is essential to prevent the spread of plant disease, especially in field and customs inspections. Leptosphaeria maculans (L. maculans) is an aggressive fungus, which causes severe phoma stem canker of Brassica napus, responsible for major yield losses of oilseed rape worldwide. In this study, CRISPR/Cas12a-based detection system and recombinase polymerase amplification (RPA) technique were employed to develop a rapid and sensitive detection method for identifying L. maculans. The involved RPA pre-amplification and CRISPR/Cas12a cleavage confer considerable sensitivity and selectivity, which can be finished within 45 min with a LOD of 4.7 genomic DNA copies. This detection system was further developed to two portable platforms, i.e., one-pot lateral flow detection and all-in-one chip lateral flow assay (AOCLFA), which integrates the lyophilized recombinase polymerase amplification (RPA) reagents and lyophilized Cas12a cleavage reagents in one tube or chip. The developed portable platforms have flexible portability and simple operation for the detection of L. maculans from plant tissues in the field. The proposed portable suitcase containing the minimum equipment, regents, and AOCLFA meets the practical needs of rapid on-site disease screening of plant fungi, port quarantine, or pathogen spreading control."

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