ABSTRACT
Encephalomyocarditis virus (EMCV) is one of the major zoonosis pathogens and can cause acute myocarditis in young pigs or reproductive failure in sows. In this study, a TaqMan-based real-time reverse transcription polymerase chain reaction (RT-PCR) assay targeting 3D gene of EMCV was developed and their sensitivities and specificities were investigated. The results indicated that the standard curve had a wide dynamic range (10(1)-10(6) copies/µL) with a linear correlation (R(2)) of 0.996 between the cycle threshold (Ct) value and template concentration. The real-time RT-PCR assay is highly sensitive and able to detect 1.4×10(2) copies/µL of EMCV RNA, as no cross-reaction was observed with other viruses. These data suggested that the real-time RT-PCR assay developed in this study will be suitable for future surveillance and specific diagnosis of EMCV-infection.
Subject(s)
Cardiovirus Infections/veterinary , Encephalomyocarditis virus/isolation & purification , Real-Time Polymerase Chain Reaction/methods , Reverse Transcriptase Polymerase Chain Reaction/methods , Swine Diseases/diagnosis , Swine Diseases/virology , Veterinary Medicine/methods , Animals , Cardiovirus Infections/virology , Cross Reactions , Sensitivity and Specificity , SwineABSTRACT
The encephalomyocarditis virus (EMCV) can cause acute myocarditis in young pigs or reproductive failure in sows. In this study, a reverse transcription loop-mediated isothermal amplification (RT-LAMP) assay was developed to detect EMCV RNA. The RT-LAMP assay was highly sensitive and able to detect 2.2 × 10(-5)ng of EMCV RNA, as no cross-reaction was observed with other viruses. The RT-LAMP assay was conducted in isothermal (62 °C) conditions within 50 min. The amplified products of EMCV could be detected as ladder-like bands using agarose gel electrophoresis. This is the first report to demonstrate the application of a one-step RT-LAMP assay for the detection of EMCV. The sensitive, specific and rapid RT-LAMP assay developed in this study can be applied widely in clinical diagnosis and field surveillance of EMCV.
