Detection of bluetongue virus in sheep by real-time RT-PCR in different production systems in San Martin, Peru

Abstract

The present study aimed to determine the prevalence of Bluetongue Virus (BTV) in sheep, by the real-time Reverse Transcription-polymerase chain reaction (RT-PCR) technique. Three hundred sixty-six sheep from the ten provinces of the Peru region were evaluated. The methodology used was the collection of blood samples from the jugular vein of the sheep, then the process of extraction and purification of RNA was carried out with the QIAmp® kit, then the reverse transcription to obtain the cDNA, and finally perform the real-time RT-PCR, for which the SuperScript III platinium One-step qRT-PCR kit was used, with the primers and probes being directed to segment 10 of the NS3 gene of BTV. The results of the real-time RT-PCR test revealed two positive sheep with a value of cycle threshold (Ct) of 35.21 and 35.57, with a prevalence of 0.54 % of BTV-positive sheep in the extensive production system, with environmental conditions that favor the development of the Culicoides vector. It is concluded that, by means of the real-time RT-PCR technique, the presence of BTV in this region of Peru is confirmed, which makes future studies necessary to determine the detection of other potential serotypes of BTV in the Peruvian Amazon in order to improve the control strategies of the disease.