Variation inBluetongue virusreal-time reverse transcription polymerase chain reaction assay results in blood samples of sheep, cattle, and alpaca

Abstract

Abstract. Bluetongue is a vector-borne viral disease that affects domestic and wild ruminants. The epidemiology of this disease has recently changed, with occurrence in new geographic areas. Various real-time quantitative reverse transcription polymerase chain reaction (real-time qRT-PCR) assays are used to detect Bluetongue virus (BTV); however, the impact of biologic differences between New World camelids and domestic ruminant samples on PCR efficiency, for which the BTV real-time qRT-PCR was initially validated are unknown. New world camelids are known to have important biologic differences in whole blood composition, including hemoglobin concentration, which can alter PCR performance. In the present study, sheep, cattle, and alpaca blood were spiked with BTV serotypes 10, 11, 13, and 17 and analyzed in 10-fold dilutions by real-time qRT-PCR to determine if species affected nucleic acid recovery and assay performance. A separate experiment was performed using spiked alpaca blood subsequently diluted in 10-fold series in sheep blood to assess the influence of alpaca blood on performance efficiency of the BTV real-time qRT-PCR assay. Results showed that BTV-specific nucleic acid detection from alpaca blood was consistently 1-2 logs lower than from sheep and cattle blood, and results were similar for each of the 4 BTV serotypes analyzed.

Diagnostic Tools for Bluetongue and Epizootic Hemorrhagic Disease Viruses Applicable to North American Veterinary Diagnosticians

“…Nucleic acid extraction methods must be evaluated for the animal source (Brito et al 2011) and the specimen chosen (Vanbinst et al 2010). EDTA blood represents the most common sample type for molecular detection of BTV (MacLachlan et al 2009); however, PCR inhibitors, such as immunoglobulin G and hemoglobin, can have significant negative effects on the sensitivity of RT-PCR-based tests (Al-Soud and Radstrom 2001).…”

Metagenomic Next-Generation Sequencing Reveal Presence of a Novel Ungulate Bocaparvovirus in Alpacas

“…Over the past couple of decades, alpacas have gained significant popularity as pets, show animals, and fiber animals in the United States, with a total of 264,587 alpacas registered in the US as of May 2019 [14]. A variety of viruses have been identified in alpacas, including bovine viral diarrhea virus, coronavirus, adenovirus, equine viral arteritis virus, rotavirus, rabies, bluetongue virus, foot-and-mouth disease virus, bovine respiratory syncytial virus, influenza A virus, bovine papillomavirus, vesicular stomatitis virus, parainfluenza-3 virus, West Nile virus, and equine herpesvirus [12,15,16,17,18,19,20]. However, BoVs have yet to be reported in alpacas.…”