科研动态

重组酶聚合酶扩增技术（recombinase polymerase amplification，RPA ）是近几年新发展并普及应用的一种核酸扩增技术，能够实现检测设备小型化，从而满足现场诊断（point-of-care testing，POCT）的需求。本研究以鲤春病毒血症病毒（spring viraemia of carp virus，SVCV） 为检测靶标，建立了SVCV的RPA检测技术，并分析了该方法的灵敏性、特异性和稳定性，评判了该技术应用于POCT的可能性。结果表明：基于RPA的检测方法灵敏度较高，检测下限可达4×103 PFU/mL，但比以Taq酶为基础的RT-PCR技术略低；特异性好，未发现对其他水生动物疫病病原，如传染性造血器官坏死病病毒（IHNV）、病毒性出血性败血症病毒（VHSV）、传染性胰腺坏死病病毒（IPNV）、传染性鲑鱼贫血病病毒（ISAV）等发生非特异性反应；稳定性良好，3次重复性试验的结果近乎一致。试验证明，该方法能够替代RT-PCR技术，应用于POCT的试剂和仪器设备研发。

Establishment and Evaluation of a Nucleic Acid Detection Technique for Spring Viraemia of Carp Virus Based on RPA

Recombinase polymerase amplification（RPA）is a kind of nucleic acid amplification method developed and widely used in recent years，which could make relevant detection equipments miniaturized to satisfy the requirements of point-of-care testing（POCT）. In this study，based on spring viraemia of carp virus（SVCV），the RPA method for detection of SVCV was established，and its sensitivity，specificity and stability were analyzed，then the possibility of applying this method to POCT was evaluated. The results showed that the method was with high sensitivity，and its lower limit detection was 4×103 PFU/mL which was slightly lower than that of Taq enzyme based RT-PCR；its specificity was good，no any non-specific reaction with the pathogens of other aquatic animal was found，such as IHNV，VHSV，IPNV and ISAV；and the results of three repeated tests were almost the same due to its good stability. In conclusion，the RPA method could be used in research of relevant instruments and reagents for POCT.

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