Detection of Avian Reovirus Infection Using a Reverse-Transcription Loop-mediated Isothermal Amplification Assay

The method of reverse transcription loop-mediated isothermal amplification (RT-LAMP) was developed for the detection of avian reovirus (ARV) infections, using primers specific to ARV S1. The amplification products were detected in three strains of ARV; whereas, none was obtained from the irrelevant controls. The sensitivity of RT-LAMP for ARV dsRNA was determined and compared with reverse transcription-polymerase chain reaction (RT-PCR) in parallel. The detection limit of RT-LAMP was 0.01 pg of dsRNA which was 10-fold more sensitive than that of RT-PCR. In addition to detecting ARV RNA in tendon tissues collected from S1133-infected chickens, RT-LAMP could also detect ARV RNA in 16 out of 23 tendon-tissue samples prepared from commercial broilers. The RT-LAMP detection results concorded 100% (16/16) with that detected by RT-PCR. This study demonstrates a speed, specificity and sensitivity RT-LAMP that is relevant for the diagnosis of ARV infection.