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A rapid simultaneous detection of duck hepatitis A virus 3 and novel duck reovirus based on RPA CRISPR Cas12a/Cas13a

文献类型： 外文期刊

作者： Zhang, Qiaoli 1 ; Yu, Guanliu 1 ; Ding, Xinli 2 ; Zhang, Kaini 1 ; Sun, Wenbo 3 ; Li, Qingmei 4 ; Yi, Yunpeng 5 ; Wang, Jianhua 6 ; Pang, Xuehui 7 ; Chen, Lei 1 ;

作者机构： 1.Shandong Normal Univ, Coll Life Sci, Shandong Prov Key Lab Anim Resistance Biol, Jinan, Shandong, Peoples R China

2.Shandong Inst Commerce & Technol, Dept Food Ind, 4516 Lvyou Rd, Jinan, Peoples R China

3.Shandong Acad Agr Sci, Inst Anim Sci & Vet Med, Shandong Key Lab Anim Dis Control & Breeding, Jinan, Shandong, Peoples R China

4.Henan Acad Agr Sci, Inst Anim Hlth Prevent & Control, Zhengzhou, Henan, Peoples R China

5.Shandong Acad Agr Sci, Inst Poultry Sci, Shandong Prov Anim & Poultry Green Hlth Prod Creat, 202 Gongyebeilu, Jinan, Shandong, Peoples R China

6.Shandong Hekangyuan Biol Breeding Co LTD, Jinan, Shandong, Peoples R China

7.Tiangong Univ, Sch Life Sci, Tianjin, Peoples R China

关键词： RPA-CRISPR Cas12a/Cas13a; Duck hepatitis a virus 3; Novel duck reovirus; Lateral flow assay

期刊名称：INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES （ 影响因子：7.7； 五年影响因子：7.7 ）

ISSN： 0141-8130

年卷期： 2024 年 274 卷

页码：

收录情况： SCI

摘要： The mixed infection of duck hepatitis A virus 3 (DHAV-3) and novel duck reovirus (NDRV) has caused significant losses to the global duck farming industry. On -site point-of-care testing of viruses plays a crucial role in the early diagnosis, prevention, and disease control. Here, we proposed an RPA-CRISPR Cas12a/Cas13a one -pot strategy (DRCFS) for rapid and simultaneous detection of DHAV-3 and NDRV. This method integrated the reaction of RPA and CRISPR Cas12a/Cas13a in a single tube, eliminating the need to open the lid during the intermediate processes and thereby avoiding aerosol contamination. On this basis, we proposed a dual RPA-CRISPR strategy coupled with a lateral flow analysis platform (DRC-LFA). This circumvented the necessity for complex instruments, enabling direct visual interpretation of results, making the test more accessible and user-friendly. Our findings demonstrated that the DRCFS method could detect DHAV-3 and NDRV at concentrations as low as 10 0 copy/ mu L, while DRC-LFA achieved limit of 10 1 copies/ mu L within 35 min. Furthermore, when DRCFS, DRC-LFA, and qPCR were employed collectively for clinical samples analysis, all three methods yielded consistent results. The specificity, sensitivity, and user-friendly of these methods rendered them invaluable for on -site virus detection.

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