Development and evaluation of TaqMan real-time PCR assay for detection of beak and feather disease virus

Černíková, Lenka; Vitásková, Eliška; Nagy, Alexander

doi:10.1016/j.jviromet.2017.02.017

Cited by 9 publications

(3 citation statements)

References 22 publications

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“…Large-scale multiple sequence alignments (LS-MSA) represent an invaluable information source for a universal diagnostic PCR design or reassessment [69][70][71]. A comprehensive analysis of sequence variability based on LS-MSAs was also critical to the presented study.…”

Section: Discussionmentioning

confidence: 99%

A universal RT-qPCR assay for “One Health” detection of influenza A viruses

et al. 2021

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The mutual dependence of human and animal health is central to the One Health initiative as an integrated strategy for infectious disease control and management. A crucial element of the One Health includes preparation and response to influenza A virus (IAV) threats at the human-animal interface. The IAVs are characterized by extensive genetic variability, they circulate among different hosts and can establish host-specific lineages. The four main hosts are: avian, swine, human and equine, with occasional transmission to other mammalian species. The host diversity is mirrored in the range of the RT-qPCR assays for IAV detection. Different assays are recommended by the responsible health authorities for generic IAV detection in birds, swine or humans. In order to unify IAV monitoring in different hosts and apply the One Health approach, we developed a single RT-qPCR assay for universal detection of all IAVs of all subtypes, species origin and global distribution. The assay design was centred on a highly conserved region of the IAV matrix protein (MP)-segment identified by a comprehensive analysis of 99,353 sequences. The reaction parameters were effectively optimised with efficiency of 93–97% and LOD95% of approximately ten IAV templates per reaction. The assay showed high repeatability, reproducibility and robustness. The extensive in silico evaluation demonstrated high inclusivity, i.e. perfect sequence match in the primers and probe binding regions, established as 94.6% for swine, 98.2% for avian and 100% for human H3N2, pandemic H1N1, as well as other IAV strains, resulting in an overall predicted detection rate of 99% on the analysed dataset. The theoretical predictions were confirmed and extensively validated by collaboration between six veterinary or human diagnostic laboratories on a total of 1970 specimens, of which 1455 were clinical and included a diverse panel of IAV strains.

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Section: Discussionmentioning

confidence: 99%

A universal RT-qPCR assay for “One Health” detection of influenza A viruses

et al. 2021

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“…19 Real-time PCR (rtPCR) assays using SYBR green or SYTO9 (intercalating fluorescent dyes) have been independently developed for BFDV, APV1, and PsHV1, 13,24 and validations of TaqMan-based rtPCR assays have also been published for BFDV in 2 reports. 3,10 Psittacine birds can be infected with multiple viruses simultaneously, as documented in numerous reports of coinfection with APV1, BFDV, and/or PsHV1. 6,15,16,25 A multiplex rtPCR assay that can test for multiple pathogens in a single reaction would reduce the cost of processing, providing additional data on presence or absence of pathogens that would otherwise not be assessed with individual tests.…”

Section: Introductionmentioning

confidence: 98%

Development and use of a triplex real-time PCR assay for detection of three DNA viruses in psittacine birds

et al. 2019

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Aves polyomavirus 1, psittacine beak and feather disease virus, and psittacid herpesvirus 1 are important pathogens of psittacine birds with the potential to cause substantial morbidity and mortality. Using publically available nucleotide sequences, we developed and validated a triplex real-time PCR (rtPCR) assay to rapidly detect these 3 viruses. The assay had high analytical sensitivity, detecting <6 copies of viral DNA per reaction, and 100% analytical specificity, showing no cross-reactivity with 59 other animal pathogens. Archived formalin-fixed, paraffin-embedded tissues from psittacine birds diagnosed at postmortem as infected with each of the viruses as well as virus-negative birds were used to validate the utility of the assay. Birds were selected for the positive cohort if they showed histologic evidence of infection (i.e., characteristic inclusion bodies in tissues); birds in the negative cohort had final diagnoses unrelated to the pathogens of interest. The triplex rtPCR assay confirmed 98% of histopathology-positive cases, and also identified subclinical infections that were not observed by histologic examination, including coinfections. Birds that tested positive only by rtPCR had significantly higher cycle threshold values compared to those with histologic evidence of infection. Positive, negative, and overall percentage agreements as well as the kappa statistic between the results of the assay and histopathology were high, demonstrating the usefulness of the assay as a tool to confirm disease diagnoses, and to improve detection of subclinical infections.

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“…Transmission mechanism of the PBFD include direct contact, fecal-oral route, vertical transmission (i.e., from the parent to the offspring), contaminated water and food, feather and skin particles [22, To this end, given very limited information on the role of asymptomatic individuals in PBFDV transmission, we aimed to investigate the temporal change in viral load in feather and fecal samples from 17 asymptomatic A. roseicollis, as feather and feces are the primary means of viral shedding. To achieve that, we developed a quantitative PCR (qPCR) assay for PBFDV DNA quantification in our infected birds since PBFDV genome is very variable with some studies showing host-specific viral sequences [5,43]. Our findings provide valuable insights into the dynamics of host-virus infections and offer crucial information for disease and conservation management.…”

Section: Introductionmentioning

confidence: 99%

Temporal characterization of the viral load of psittacine beak and feather disease virus in rosy-faced lovebirds (Agapornis roseicollis)

Lam,

Kei Poon,

Sin

2024

Preprint

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Psittacine beak and feather disease virus (PBFDV) is a widespread and highly pathogenic virus in parrots (Psittaciformes), threatening both captive and wild populations over the world. The disease typically presents with feather and beak abnormalities, along with possible immune system suppression. No cure or commercialized vaccine is currently available. Our understanding of the Psittacine beak and feather disease often come from infected individuals with visible symptoms. Limited knowledge exists regarding the pathology and role of asymptomatic individuals in disease transmission. Asymptomatic individuals could shed virus in their crop secretion, feces, or feathers. In this study, we investigated the temporal change in viral load in feather and fecal samples from 17 asymptomatic rosy-faced lovebirds (Agapornis roseicollis). We developed a qPCR assay for PBFDV viral load quantification in the studied lovebirds. Our results showed that most of the individuals had very low viral load, while three individuals with high viral load at the beginning of the experiment were observed to exhibit a decreasing trend in viral load in both fecal and feather samples. Surprisingly, the viral load in an individual can drop from a high level to an undetectable level within three months, which is contrary to the prevailing notion that the disease is highly lethal with few reports of complete recovery. We also showed that viral load in feathers was higher than in feces. Our study provides valuable insights into the infection dynamics of PBFDV in asymptomatic individuals and contribute to the understanding of disease transmission in parrots.

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Development and evaluation of TaqMan real-time PCR assay for detection of beak and feather disease virus

Cited by 9 publications

References 22 publications

A universal RT-qPCR assay for “One Health” detection of influenza A viruses

A universal RT-qPCR assay for “One Health” detection of influenza A viruses

Development and use of a triplex real-time PCR assay for detection of three DNA viruses in psittacine birds

Temporal characterization of the viral load of psittacine beak and feather disease virus in rosy-faced lovebirds (Agapornis roseicollis)

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