Recombinase-Aided Amplification Coupled with Lateral Flow Dipstick for Efficient and Accurate Detection of Porcine Parvovirus.

Yihong He,Jinding Chen,Hongxing Ding,Jindai Fan,Lin Yi,Shuangqi Fan,Wenxian Chen

doi:10.3390/life11080762

Abstract

Porcine parvovirus (PPV) infection is the primary cause of SMEDI (stillbirth; mummification; embryonic death; infertility) syndrome, which is a global burden for the swine industry. Thus, it is crucial to establish a rapid and efficient detection method against PPV infection. In the present work, we developed a recombinase-aided amplification (RAA) assay, coupled with a lateral flow dipstick (LFD), to achieve an amplification of PPV DNA at 37 °C within 15 min. The detection limits of PPV RAA-LFD assay were 102 copies/μL recombinant plasmid pMD19-T-VP1, 6.38 × 10−7 ng/μL PPV DNA, and 10−1 TCID50/mL virus, respectively. This method was highly specific for PPV detection with no cross-reactivity for other swine pathogens. In contrast to polymerase chain reaction (PCR), the PPV RAA-LFD assay is more sensitive and cost-saving. Hence, the established PPV RAA-LFD assay provided an alternative for PPV detection, especially in resource-limited regions.

Highlights

Porcine parvovirus (PPV) is a small non-enveloped, single-stranded DNA virus belonging to the Parvoviridae family, the Parvovirinae subfamily and the Ungulate protoparvovirus I species [1]
It leads to a financially damaging disease known as PPV infection, characterized by SMEDI syndrome [1,2]
ETvhaliurtayti-oenigohftCsluinsipcealctSeadmspalmesples stored in our laboratory were simultaneously tested withTthheirtPyP-eVigRhAt Asu-sApGecEteadsssaaymapnldesthsteoPrePdVinPCouRr-AlaGboEraastosaryy wtoeerveasliumautelttahneeofeuassliybitleistyteodf wthiethcltihneicPaPl aVpRpAlicAa-tiAoGn,Ereasspsaeyctiavnedlyt.he PPV polymerase chain reaction (PCR)-agarose gel electrophoresis (AGE) assay to evaluate the feasibility of the clinical application, respectively

Summary

Introduction

Porcine parvovirus (PPV) is a small non-enveloped, single-stranded DNA virus belonging to the Parvoviridae family, the Parvovirinae subfamily and the Ungulate protoparvovirus I species [1]. It leads to a financially damaging disease known as PPV infection, characterized by SMEDI (stillbirth; mummification; embryonic death; infertility) syndrome [1,2]. Isolated from sows [4], PPV is endemic worldwide, seriously influencing herd profitability due to the reproduction failure it causes [5], especially primiparous sows. There is an urgent need to develop an efficient and accurate detection system for PPV detection

Methods

Results

Conclusion