Development of a real time reverse transcription loop-mediated isothermal amplification method (RT-LAMP) for detection of a novel swine acute diarrhea syndrome coronavirus (SADS-CoV)

Abstract

A novel swine acute diarrhea syndrome Coronavirus (SADS-CoV) that causes severe diarrhea in suckling piglets was identified in southern China in 2017. A simple and rapid detection test was developed for this virus using real-time RT-LAMP based on the conserved N gene of the virus. The method had a detection limit of 1.0 × 10 copies/μL with no cross-reactions with classical swine fever virus, porcine and respiratory syndrome virus NA, porcine and respiratory syndrome virus EU, transmissible gastroenteritis coronavirus, foot and mouth disease virus, porcine epidemic diarrhea virus (S-INDEL and non-S-INDEL), swine influenza virus subtype H1N1, porcine circovirus type 2, seneca valley virus, porcine parvovirus, porcine deltacoronavirus and rotavirus. This method was also reproducible. Twenty of 24 clinical samples were identified as SADS-CoV RNA-positive by the real-time RT-LAMP and the results were consistent with that of the real time RT-PCR method. This new method for detecting SADS-CoV is specific and sensitive for the detection of SADS-CoV.

Development of a reverse transcription-loop-mediated isothermal amplification as a rapid early-detection method for novel SARS-CoV-2

“…To evaluate the time efficiency of the developed assay, optimization of the RT-LAMP reaction was done at varying reaction time points (10,20,30,40,50, and 60 min) using the determined limit concentration of viral RNA. Ten repeats of the RT-LAMP reaction were performed using the determined viral RNA concentrations with incubations of 30 and 60 min.…”

“…The specificity and sensitivity of this method is generally comparable to those of the conventional PCR and qRT-PCR. The LAMP assays merged with reverse transcription steps have been developed for the detection of RNA viruses, including SARS-CoV, MERS-CoV, influenza, and other respiratory viruses [26][27][28][29][30][31][32].…”

Development of a real-time loop-mediated isothermal amplification assay for detection of porcine circovirus 3

“…Six porcine viruses were used for specificity testing of PCV3. These viruses were classical swine fever virus (CSFV) C strain, foot-and-mouth disease virus (FMDV), porcine circovirus 2 (PCV2) LG vaccine strain, porcine epidemic diarrhoea virus (PEDV), porcine respiratory and reproductive syndrome virus (PRRSV), and pseudorabies virus (PRV) [13,15].…”

“…Compared to conventional LAMP assays, this method avoids visible error, enables quantitative detection and is more suitable for multi-sample analysis. Because of its simple reaction conditions and ease of use, the realtime LAMP method has been adapted for the detection and diagnosis of a variety of pathogens [12,13]. In this study, a real time RT-LAMP assay was developed for the rapid diagnosic of PCV3.…”

Significant inhibition of re-emerged and emerging swine enteric coronavirus in vitro using the multiple shRNA expression vector

“…Swine acute diarrhea syndrome coronavirus (SADS-CoV) is a novel coronavirus which was first reported in southern China in 2017. It also can cause severe diarrhea disease in newborn piglets (Fu et al, 2018;Pan et al, 2017;Wang et al, 2018aWang et al, , 2018bXu et al, 2018;Zhou et al, 2018aZhou et al, , 2018b. Similar to other SECoVs, such as transmissible gastroenteritis virus (TGEV), these re-emerged and emerging SECoV diseases are age-dependent with high morbidity and mortality in neonatal pigs.…”