中国兽医科学2024，Vol.54Issue(5)：595-601,7.DOI:10.16656/j.issn.1673-4696.2024.0099
传染性皮下和造血器官坏死病毒RPA与CRISPR-Cas12a可视化快速检测方法的建立
Establishment of rapid visual detection methods for infectious hypodermal and hematopoietic necrosis virus based on RPA and CRISPR-Cas12a
摘要
Abstract
After screening and optimization,two visual rapid detection methods,the RPA-Cas12a fluorescent detection and lateral flow dipstick,for infectious hypodermal and hematopoietic necrosis virus(IHHNV)were established by designing and synthesizing multiple sets of recombinase polymerase amplification primers and specific CRISPR RNA based on the conserved regions of the IHHNV gene se-quences.The results demonstrated that both visual detection methods could detect IHHNV after a con-stant temperature reaction at 37℃for 40 min.There was no cross-reactivity with white spot syndrome virus,Enterocytozoon hepatopenaei and decapod iridescent virus 1.The method exhibited high sensiti-vity,with a detection limit of 10 copies/μL.When applied to test 10 positive clinical samples of IHH-NV,the conformity rate was 100%.Overall,the established fluorescence and paper strip visual detec-tion methods are simple to operate,highly sensitive,and suitable for regular monitoring and prelimi-nary screening of IHHNV in shrimp farming facilities or basic laboratories.关键词
传染性皮下和造血器官坏死病毒/重组酶聚合酶扩增/CRISPR-Cas12a/检测方法Key words
infectious hypodermal and hematopoietic necrosis virus/recombinase polymerase am-plification/CRISPR-Cas12a/detection method分类
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周傲白雪,陈轩,徐博文,沙才华,邵建宏,赵福振,廖秀云,黄海超,蒋晓霞,罗宝正..传染性皮下和造血器官坏死病毒RPA与CRISPR-Cas12a可视化快速检测方法的建立[J].中国兽医科学,2024,54(5):595-601,7.基金项目
广东省重点领域研发计划项目(2022B1111030001) （2022B1111030001）
拱北海关科研项目(2023GK009) （2023GK009）
