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| Rapid Detection of Decapod iridescent virus 1 (DIV1) by Recombinase Polymerase Amplification |
| YIN Weili1, WU Wei2, LIU Xiaojing3, MA Xiaoling4, HAN Wei5, WU Ningning5, CHEN Yanping1 |
1. Yantai Customs Technical Center, Yantai 264000, China;
2. Wuhan Agricultural Testing Center, Wuhan 430016, China;
3. Rongcheng Customs, Rongcheng 264300, China;
4. Qingdao Guofeng Pharmaceutical Co., Ltd., Shanghai Pharmaceutical Group, Qingdao 266000, China;
5. Qingdao Customs Technical Center, Qingdao 266000, China |
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Abstract A rapid detection of Decapod iridescent virus 1 (DIV1) was established based on the recombinase polymerase amplification technology(RPA)using primers designed according to ATPase gene (ORF114R), with the optimal amplification temperature of the RPA method established in this study at 30 ℃, and completing within 20 min. The specificity test results indicated that there was no cross-reaction between the RPA method established in this study and the other shrimp virus infection. The sensitivity test revealed that the detection limit of this method was 7 copies/μL, more sensitive than the nested-PCR method. Moreover, a total of 509 clinical samples were detected by the RPA method and the PCR method, respectively. The results showed that the RPA method effectively detected the weakly positive samples from the PCR method, which was used in clinical detection. The findings indicated that the RPA method in this study was featured by fast, simple, specific and sensitive for local laboratory and on-site inspection application.
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Received: 28 October 2019
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