Detecção do vírus da bronquite infecciosa das galinhas e do metapneumovírus aviário utilizando uma reação de transcrição reversa com reação em cadeia pela polimerase duplex

A duplex RT-PCR assay is reported for the simultaneous detection of avian infectious bronchitis virus (IBV) and avian metapneumovirus (aMPV), the causative agents of major diseases in poultry. The duplex RT-PCR assay optimized showed a detection limit of 10-3 (101 EID50/50m L for IBV and 100.5 EID50/50m L for aMPV, respectively when two viruses were mixed and 10-1 for each one separated (103 EID50/50m L for IBV and 102.5 EID50/50m L for aMPV, respectively. It was specific, sensitive and applicable for the rapid detection of these viruses in clinical samples.  Descreve-se um ensaio de duplex RT-PCR assay para a detecção simultânea do vírus da bronquite infecciosa das galinhas (IBV) e do metapneumovirus aviário (aMPV), agentes etiológicos de doenças de elevada importância em avicultura. A duplex RT-PCR otimizada mostrou um limiar de detecção de 10-3 (101 EID50/50m L para IBV e 100.5 EID50/50m L para aMPV, respectivamente, quando da combinação dos dois vírus e 10-1 para cada um dos vírus em separado(103 EID50/50m L para IBV e 102.5 EID50/50m L para aMPV, respectivamente. O ensaio foi demonstrado como específico, sensível e aplicável à rápida detecção destes vírus em amostras clínicas.

Survey of feline leukemia virus and feline coronaviruses in captive neotropical wild felids from southern Brazil

Abstract: A total of 57 captive neotropical felids (one Leopardus geoffroyi, 14 Leopardus pardalis, 17 Leopardus\ud wiedii, 22 Leopardus tigrinus, and three Puma yagouaroundi) from the Itaipu Binacional Wildlife Research Center\ud (Refu´gio Bela Vista, Southern Brazil) were anesthetized for blood collection. Feces samples were available for 44\ud animals, including one L. geoffroyi, eight L. pardalis, 14 L. wiedii, 20 L. tigrinus, and one P. yagouaroundi. Total\ud DNA and RNA were extracted from blood and feces, respectively, using commercial kits. Blood DNA samples\ud were evaluated by polymerase chain reaction (PCR) for feline leukemia virus (FeLV) proviral DNA, whereas\ud reverse transcriptase–PCR was run on fecal samples for detection of coronavirus RNA. None of the samples were\ud positive for coronaviruses. A male L. pardalis and a female L. tigrinus were positive for FeLV proviral DNA, and\ud identities of PCR products were confirmed by sequencing. This is the first evidence of FeLV proviral DNA in these\ud species in Southern Brazil