Development of a TaqMan-based real-time reverse transcription polymerase chain reaction assay for the detection of encephalomyocarditis virus.

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.

Rapid detection of encephalomyocarditis virus by one-step reverse transcription loop-mediated isothermal amplification method.

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.