Using loop-mediated isothermal amplification for visual detection of porcine parvovirus 7

利用环介导等温扩增技术进行猪细小病毒7的视觉检测

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Abstract

We developed and optimized a loop-mediated isothermal amplification (LAMP)-based method to detect porcine parvovirus 7 (PPV7). After using three pairs of specific primers to amplify PPV7 isothermally at 62 °C for 40 min, the amplified product was mixed with SYBR Green I, after which the sample turned green. The method detected PPV7 at concentrations as low as 40 copies/μL, and the sensitivity was consistent with that of nested polymerase chain reaction (PCR) analysis, which was tenfold higher than that of conventional PCR. No cross-reactivity occurred with porcine parvovirus 1, porcine circovirus type 3, porcine circovirus type 2, porcine pseudorabies virus, porcine epidemic diarrhea virus, or porcine reproductive and respiratory syndrome virus. Simultaneous analysis of 76 clinical samples was performed using LAMP, conventional PCR, and nested PCR. The results showed that our method is simple, rapid, sensitive, and specific for the rapid diagnosis of PPV7 in pig farms. SUPPLEMENTARY INFORMATION: The online version of this article (10.1007/s13205-020-02623-5) contains supplementary material, which is available to authorized users.

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