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Rapid detection of giant salamander iridovirus by cross-priming amplification

文献类型： 外文期刊

作者： Zhang, Hao 1 ; Wang, FangYu 2 ; Jia, Rui 3 ; Fan, Jianming 5 ; Hao, Junfang 2 ; Du, Jiaru 2 ; Niu, Yan 2 ; Han, Sifang 1 ; D 1 ;

作者机构： 1.Henan Agr Univ, Sch Life Sci, Zhengzhou 450000, Henan, Peoples R China

2.Henan Acad Agr Sci, Key Lab Anim Immunol, Zhengzhou 450000, Henan, Peoples R China

3.Zhengzhou Univ, Sch Life Sci, Zhengzhou 450000, Henan, Peoples R China

4.Sichuan Agr Univ, Coll Vet Med, Avian Dis Res Ctr, Chengdu 611830, Sichuan, Peoples R China

5.Zhengzhou Univ, Sch Publ Hlth, Zhengzhou 450000, Henan, Peoples R China

6.Henan Agr Univ, Zhengzhou 450000, Henan, Peoples R China

关键词： Giant salamander iridovirus; Cross-Priming amplification; Nucleic acid strip; Rapid detection

期刊名称：JOURNAL OF VIROLOGICAL METHODS （ 影响因子：2.014； 五年影响因子：2.001 ）

ISSN： 0166-0934

年卷期： 2019 年 274 卷

页码：

收录情况： SCI

摘要： Giant salamander iridovirus (GSIV) belongs to the epizootic genus Ranavirus, and is the cause of epidemic diseases associated with high mortality and great losses to artificial breeding and farming. Here, we established a simple, accurate, and reliable cross-priming amplification (CPA) method to detect GSIV. The CPA assay targets the major caspid protein gene of the GSIV genome to design crossing primer pairs, and the reaction conditions were optimized, including optimal concentrations of the primers, betaine, dNTPs, Mg2+, and Bst DNA polymerase, and reaction conditions. The sensitivity was shown to be 10 times higher than that of conventional polymerase chain reaction (PCR), and the specificity was 100%. The results were identified on nucleic acid strips within 3-5 min. Application of the CPA and PCR to 54 samples of giant salamander showed a positive rate of 72.22% and 74.07%, respectively, demonstrating high coincidence (94.44%, kappa = 8.7, P < 0.0001). The sensitivity of the CPA assay was 97.50% and the specificity was 92.86%. Thus, the CPA assay is as effective as conventional PCR, but with added practical advantages of simplicity and an almost instrument-free platform, which will be useful for both laboratories and giant salamander farms.

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