Rapid and sensitive detection of Senecavirus A by reverse transcription loop-mediated isothermal amplification combined with a lateral flow dipstick method.

Jinhui Li,Jingfeng Xiong,Xia Zhou,Guangbin Si,Shuaifei Xu,Li Yu,Jian Shi,Bowen Liu,Dongsheng He,Weifang Liang,Ralph A Tripp

doi:10.1371/journal.pone.0216245

Abstract

Senecavirus A (SVA) is a critical pathogen causing vesicular lesions in sows and acute death of newborn piglets, resulting in very large economic losses in the pig industry. To restrict the transmission of SVA, an establishment of an effective diagnostic method is crucial for the prevention and control of the disease. However, traditional detection methods often have many drawbacks. In this study, reverse transcription loop-mediated isothermal amplification (RT-LAMP) was combined with a lateral flow dipstick (LFD) to detect SVA. The resulting RT-LAMP-LFD assay was performed at 60°C for 50 min and then directly judged on an LFD visualization strip. This method shows high specificity and sensitivity to SVA. The detection limit of RT-LAMP was 4.56x10-8 ng/μL RNA, approximately 11 copies/μL RNA, and it was 10 times more sensitive than RT-PCR. This detection method’s positive rate for clinical samples is comparable to that of RT-PCR. This method is time saving and highly efficient and is thus expected to be used to diagnose SVA infections in this field.

Highlights

Senecavirus A (SVA), formerly known as Seneca Valley virus (SVV)[1], is the only recognized species of the genus Senecavirus within the family Picornaviridae[2], and it was first isolated in 2002[3]
The products were detected by 1.2% agarose gel electrophoresis, calcein assay and lateral flow dipstick (LFD) assay to verify the feasibility of the reverse transcription loop-mediated isothermal amplification (RT-Loop-mediated isothermal amplification (LAMP)) method
The results show that the SVA RT-LAMP-LFD detection method is feasible

Summary

Introduction

Senecavirus A (SVA), formerly known as Seneca Valley virus (SVV)[1], is the only recognized species of the genus Senecavirus within the family Picornaviridae[2], and it was first isolated in 2002[3]. SVA is clinically characterized by vesicular ulcers in the coronary arteries or nostrils [4]. It is difficult to differentiate from foot-and-mouth disease (FMD), swine vesicular disease (SVD), and vesicular stomatitis (VS) in clinical symptoms[5]. SVA is a small, nonenveloped RNA virus with a single sense strand 27–30 nm in diameter. The viral genome is approximately 7.3 kb in length and consists of a 5’ noncoding region (UTR), a 3’ noncoding region (UTR) and a large open reading frame[3].

Methods

Results

Conclusion