Rapid and sensitive detection of mink circovirus by recombinase polymerase amplification

文献类型： 外文期刊

第一作者： Ge, Junwei

作者： Ge, Junwei;Shi, Yunjia;Cui, Xingyang;Gu, Shanshan;Ge, Junwei;Zhao, Lili;Chen, Hongyan

作者机构：

关键词： Mink circovirus; Molecular diagnosis; Nested PCR; Recombinase polymerase amplification

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： 2018 年 256 卷

页码：

收录情况： SCI

摘要： To date, the pathogenic role of mink circovirus (MiCV) remains unclear, and its prevalence and economic importance are unknown. Therefore, a rapid and sensitive molecular diagnosis is necessary for disease management and epidemiological surveillance. However, only PCR methods can identify MiCV infection at present. In this study, we developed a nested PCR and established a novel recombinase polymerase amplification (RPA) assay for MiCV detection. Sensitivity analysis showed that the detection limit of nested PCR and RPA assay was 10(1) copies/reaction, and these methods were more sensitive than conventional PCR, which has a detection limit of 10(5) copies/reaction. The RPA assay had no cross-reactivity with other related viral pathogens, and amplification was completed in less than 20 min with a simple device. Further assessment of clinical samples showed that the two assays were accurate in identifying positive and negative conventional PCR samples. The detection rate of MiCV by the RPA assay in clinical samples was 38.09%, which was 97% consistent with that by the nested PCR. The developed nested PCR is a highly sensitive tool for practical use, and the RPA assay is a simple, sensitive, and potential alternative method for rapid and accurate MiCV diagnosis.

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