外来人侵物种北美刺CRISPR鉴定方法的建立
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中图分类号：S412;S451;Q948.13 文献标志码：A 文章编号：1003-935X（2026）01-0046-07
Establishment of CRISPR - Based Identification Method for Invasive Species Solanum carolinense
SHAN Luyue²，WU Jing1 ，LI Jinggan1 ，MA Yuanwei³，SONG Xiaoling²，FU Jianguo 1 （1.Nanjing Customs Laboratory for Animal，Plant and Food Inspection and Quarantine，Nanjing 210o19，China; 2.NanjingAgiculturalUnivrsityNanjing20o5，hina；3.lashGeneLifeTechnologiesCo.，td.Nanjing2ina）
Abstract：Solanum carolinense a globall noxious invasive weed that poses a significant threat to agricultural production and ecological securityworldwide. Themorphological identification of S .carolinense seeds，which are often mixed in importedgrainsin China，isexceptionalldificult.Moreover，arapidon-site molecular biology method forits identificationhasnotyetbeenestablished，making ithardtomeetthedualrequirementsofcustomsforswift clearance and precise quarantine. In this study，specific DNA sequences of S . carolinense were screened through whole - genome sequencing and comparativeanalysis，leading to the development of arapid CRISPR-Cas12a-basedassay for S carolinense detection.This method integrates enzymatic recombinase amplification （ERA）with the trans -cleavage activity of the CRISPR-Casl2asystem，enabling high-sensitivityrecognition of target DNA.The outcomescould be visualized either byfluorescencesignal detectionorimmunochromatographictest strips，withaconsistentlower detection limit of 2. 1× 101 copies/ μL .The entire process is isothermal，and from sample processing to result interpretation，itcould becompletedwithin 3O minutes.Thisassayofers theadvantages ofhighspecificity，high sensitivity，rapidity，simplicity，andvisual resultreadout.Itefectivelyaddresses limitationsoftraditional quarantine identification technologies，providing technical support for rapidon-site screening at portsand enhancing China’s national biosecurity defense.
Key Words：Solanum carolinense L.；rapid identification； CRISPR;CRISPR-Cas12a;system
北美刺茄（SolanumcarolinenseL.）原产于美国东南部，现已扩散至美国全境1，且广泛分布于全球36个国家和地区[2]，在奥地利、日本等的分布集中于港口、交通枢纽及农田等区域[3-4]。（剩余7305字）