Validation of a High-Throughput Microfluidic Real-Time PCR for the Detection of Vector-Borne Agents in Wild Birds from the Brazilian Pantanal.

Abstract

Despite numerous studies on haemosporidians in wild birds from Brazil, the presence of other vector-borne agents (VBA) such asspp.,spp., and Onchocercidae filariids in avian hosts remains largely unknown. The low occurrence of these VBAs might be due to the low sensitivity of traditional molecular techniques. The microfluidic real-time PCR assay, known for its high sensitivity, has emerged as a promising method to detect and study the occurrence and diversity of VBAs in both arthropod vectors and vertebrate hosts. To validate previously and standardize newly designed microfluidic real-time PCR protocols, selected positive avian blood DNA samples forspp.,spp., haemosporidians, and filariids were used. The molecular occurrence rates for the selected VBAs were 18.2% forspp., 0.36% forspp., 6.2% forspp., 4.7% forspp., and 6.5% for Onchocercidae filariids. Thespp.B sequence detected in aclustered with, whereas thespp.sequence detected in aclustered with. While Onchocercidae filariid1 sequences were detected in specimens of,andgrouped withspp., one sequence detected inwas ancestral to the clade comprisingspp. andspp. High-throughput microfluidic real-time PCR assay can be used for screening VBAs in avian hosts from South America, but new primers/probe sets should be designed for VBA genotypes present in Brazil.