Optimizing the nested PCR method for Decapod iridescent virus 1 (DIV1) targeting ATPase gene by reselecting the inner primers
文献类型： 外文期刊
作者： Xing, Jing-Yi 1 ; Li, An-Qi 3 ; Guo, Xiao-Meng 1 ; Wang, Meng 1 ; Guan, Xin 1 ; Qiu, Liang 1 ; Zhang, Qing-Li 1 ; Huang, Jie 1 ;
作者机构： 1.Chinese Acad Fishery Sci, Yellow Sea Fisheries Res Inst, State Key Lab Mariculture Biobreeding & Sustainabl, Lab Marine Fisheries Sci & Food Prod Proc,Laoshan, Qingdao 266071, Peoples R China
2.Qingdao Marine Sci & Technol Ctr, Lab Marine Fisheries Sci & Food Prod Proc, Qingdao 266237, Shandong, Peoples R China
3.Tianjin Agr Univ, Coll Aquaculture, Tianjin 300384, Peoples R China
4.Shanghai Ocean Univ, Coll Fisheries & Life Sci, Shanghai 201306, Peoples R China
5.Ludong Univ, Sch Agr, Yantai 264025, Peoples R China
关键词： Decapod iridescent virus 1; Shrimp hemocyte iridescent virus; Cherax quadricarinatus iridovirus; Nested PCR; ATPase
期刊名称：JOURNAL OF INVERTEBRATE PATHOLOGY （ 影响因子：2.4； 五年影响因子：3.0 ）
ISSN： 0022-2011
年卷期： 2024 年 207 卷
页码：
收录情况： SCI
摘要： DIV1 has the characteristics of fast transmission and a broad host range. Its infection leads to a high mortality rate, posing a serious threat to the global crustacean aquaculture industry. In order to increase the accuracy of DIV1 detection and reduce the difficulty of result interpretation, this study modified the original nested PCR method targeting the DIV1 ATPase gene. The internal primers for the nested PCR were redesigned to produce a 338 bp amplification product in the second step PCR, effectively distinguishing the target band from primer dimers. The newly established nested PCR method exhibits strong specificity and high sensitivity, with a detection limit as low as 1.37 x 101 copies/reaction. The developed nested PCR assay provides new technical support for the accurate detection of DIV1 in global crustacean aquaculture.
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