Journal of Shandong University (Health Sciences) ›› 2023, Vol. 61 ›› Issue (10): 74-82.doi: 10.6040/j.issn.1671-7554.0.2023.0640

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Establishment and evaluation of a recombinase-aided amplification for the detection of human adenovirus type 4

LIU Qingqing1,2, WANG Ningning1,2, CHENG Jun2, ZHOU Huajun2, CHE Feihu2, SUN Qingyang2, WANG Yue2, ZHANG Yingjie1, DAI Yuzhu2   

  1. 1. School of Laboratory Medicine, Bengbu Medical College, Bengbu 233000, Anhui, China;
    2. Department of Clinical Research, The 903rd Hospital of the Peoples Liberation Army, Hangzhou 310013, Zhejiang, China
  • Published:2023-11-08

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Abstract: Objective To establish a simple and rapid method for human adenovirus type 4(HAdV-4)detection with recombinase-aided amplification(RAA). Methods The specific primers and probes were designed based on the Hexon gene of HAdV-4. The constructed positive plasmid standard was used for RAA under different temperatures(39 ℃, 40 ℃, 42 ℃)to select the best reaction temperature, and the sensitivity, repeatability, and specificity of the method were tested. A total of 100 clinical samples were detected with real-time RAA, and the results were compared with those of real-time PCR to evaluate the detection performance of real-time RAA. Results The optimal temperature for real-time RAA was 42 ℃, and the detection limit was 101 copies/μL. When the concentration of the plasmid standard was 101-104 copies/μL, the intra-assay coefficient of variation was less than 5%, showing good repeatability. The results of specificity showed that the method accurately detected HAdV-4 without cross-reactivity with other subtypes or other pathogens. Real-time PCR and real-time RAA of the 100 samples showed the coincidence rate was 100%. Conclusion A specific, simple and rapid assay for HAdV-4 detection is developed.

Key words: Human adenovirus type 4, Recombinase-aided amplification, Hexon gene, Rapid detection, Molecular diagnostic techniques

CLC Number: 

  • R446.5
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