scholarly journals Novel Avian Bornavirus in a Nonpsittacine Species (Canary; Serinus canaria) with Enteric Ganglioneuritis and Encephalitis

2009 ◽  
Vol 83 (21) ◽  
pp. 11367-11371 ◽  
Author(s):  
Herbert Weissenböck ◽  
Karin Sekulin ◽  
Tamás Bakonyi ◽  
Sandra Högler ◽  
Norbert Nowotny
Keyword(s):  
Sequence Analysis ◽  
Reverse Transcription ◽  
Avian Species ◽  
Serinus Canaria ◽  
Viral Dissemination ◽  
Reverse Transcription Pcr ◽  
The Family ◽  
Avian Bornavirus ◽  
Proventricular Dilatation Disease ◽  
Unique Genotype

ABSTRACT A canary bird (Serinus canaria) died with nonsuppurative ganglioneuritis of the proventriculus and gizzard and encephalitis, lesions comparable to proventricular dilatation disease (PDD) of psittacine birds. Recently, several genotypes of a novel avian bornavirus have been linked to PDD. In the canary, bornaviral antigen was detected by immunohistochemistry in both neural and extraneural tissues. The widespread viral dissemination was confirmed by reverse transcription-PCR. Sequence analysis revealed a unique genotype of avian bornavirus. This observation suggests that bornaviruses are natural pathogens of several avian species and that the family Bornaviridae comprises more viral genotypes (or viral species) than previously assumed.

scholarly journals Analysis of Naturally Occurring Avian Bornavirus Infection and Transmission during an Outbreak of Proventricular Dilatation Disease among Captive Psittacine Birds

2009 ◽  
Vol 84 (4) ◽  
pp. 2176-2179 ◽  
Author(s):  
Amy L. Kistler ◽  
Jeanne M. Smith ◽  
Alexander L. Greninger ◽  
Joseph L. DeRisi ◽  
Don Ganem
Keyword(s):  
Western Blotting ◽  
Reverse Transcription ◽  
Fatal Case ◽  
Natural Conditions ◽  
Reverse Transcription Pcr ◽  
Naturally Occurring ◽  
Avian Bornavirus ◽  
Proventricular Dilatation Disease

ABSTRACT A proventricular dilatation disease (PDD) outbreak provided the opportunity to investigate the transmissibility of avian Bornavirus (ABV) and its linkage to PDD under natural conditions. Upon exposure to a bird with a fatal case of PDD, 10 birds became symptomatic and died. ABV2 RNA was recovered from available tissues. Further screening revealed that 12/46 exposed birds were ABV2+. Three chicks boarded at this aviary developed PDD. They harbored the same ABV2 isolate and transmitted it to five of eight chicks in their home aviary. These findings demonstrate that ABV infection precedes the development of PDD. ABV-specific Western blotting and reverse transcription-PCR indicate that ABV2 is not strictly neurotropic.

scholarly journals Rapid and Sensitive Detection of Noroviruses by Using TaqMan-Based One-Step Reverse Transcription-PCR Assays and Application to Naturally Contaminated Shellfish Samples

2005 ◽  
Vol 71 (4) ◽  
pp. 1870-1875 ◽  
Author(s):  
Narayanan Jothikumar ◽  
James A. Lowther ◽  
Kathleen Henshilwood ◽  
David N. Lees ◽  
Vincent R. Hill ◽  
...  
Keyword(s):  
Reverse Transcription ◽  
Nested Pcr ◽  
Rt Pcr ◽  
Reverse Transcription Pcr ◽  
Specificity And Sensitivity ◽  
Routine Monitoring ◽  
The Family ◽  
One Step ◽  
Stool Specimens ◽  
Pcr Assays

ABSTRACT Noroviruses (NoV), which are members of the family Caliciviridae, are the most important cause of outbreaks of acute gastroenteritis worldwide and are commonly found in shellfish grown in polluted waters. In the present study, we developed broadly reactive one-step TaqMan reverse transcription (RT)-PCR assays for the detection of genogroup I (GI) and GII NoV in fecal samples, as well as shellfish samples. The specificity and sensitivity of all steps of the assays were systematically evaluated, and in the final format, the monoplex assays were validated by using RNA extracted from a panel of 84 stool specimens, which included NoV strains representing 19 different genotypes (7 GI, 11 GII, and 1 GIV strains). The assays were further validated with 38 shellfish cDNA extracts previously tested by nested PCR. Comparison with a recently described real-time assay showed that our assay had significantly higher sensitivity and was at least as sensitive as the nested PCR. For stool specimens, a one-step duplex TaqMan RT-PCR assay performed as well as individual genogroup-specific monoplex assays. All other enteric viruses examined were negative, and no cross-reaction between genogroups was observed. These TaqMan RT-PCR assays provide rapid (less than 90 min), sensitive, and reliable detection of NoV and should prove to be useful for routine monitoring of both clinical and shellfish samples.

scholarly journals Identification of Encephalomyocarditis Virus in Clinical Samples by Reverse Transcription-PCR Followed by Genetic Typing Using Sequence Analysis

1998 ◽  
Vol 36 (12) ◽  
pp. 3463-3467 ◽  
Author(s):  
H. Vanderhallen ◽  
F. Koenen
Keyword(s):  
Sequence Analysis ◽  
Reverse Transcription ◽  
Molecular Techniques ◽  
Encephalomyocarditis Virus ◽  
Clinical Samples ◽  
Nucleotide Sequence Analysis ◽  
Rt Pcr ◽  
Reverse Transcription Pcr ◽  
Field Samples ◽  
Pcr Targeting

The objective of the present study was to gain a better understanding of the epidemiology of encephalomyocarditis virus (EMCV) infections in pigs by applying molecular techniques. The diagnostic potential of a reverse transcription-PCR (RT-PCR) targeting 286 nucleotides at the 3′ end of the gene which encodes the viral polymerase was assessed with experimental and field samples. In addition, the use of the amplified sequences for an epidemiological study was evaluated. The heart was clearly shown to be the most suitable organ. The detection limit was determined to be 1 viral particle in 100 mg of heart tissue. The sensitivity and specificity of the assay on the basis of the results obtained in this study were 94 and 100%, respectively. Phylogenetic analysis of the amplified sequences classified EMCVs in two distinct lineages. Group A consists of the reference strain ATCC 129B, all isolates collected between 1991 and 1994 in Belgium in association with reproductive failure, and all Greek isolates. All Belgian isolates collected since the first isolation of EMCV in relation to myocardial failure in fatteners in Belgium group together with the isolates from Cyprus (1996 and 1997), Italy (1986 to 1996), and France (1995) in group B irrespective of their pathogenicity. The analyzed part of the 3D gene differed by 13.0% between Groups A and B. In contrast to the sequence homogeneity of the Belgian isolates collected between 1991 and 1994, molecular diversity, which ranged between 0 and 2%, was observed among the Belgian isolates collected in 1995 and 1996. Among all Greek isolates the diversity ranged between 1 and 8%. However, this diversity does not seem to reflect geographical links between the outbreaks. A RT-PCR for the rapid and specific diagnosis of EMCV in a variety of clinical samples followed by nucleotide sequence analysis proved to be valuable for molecular epidemiological studies.

scholarly journals Detection and Identification of Mammalian Reoviruses in Surface Water by Combined Cell Culture and Reverse Transcription-PCR

2001 ◽  
Vol 67 (7) ◽  
pp. 3016-3020 ◽  
Author(s):  
Michael L. Spinner ◽  
George D. Di Giovanni
Keyword(s):  
Cell Culture ◽  
Surface Water ◽  
Sequence Analysis ◽  
Reverse Transcription ◽  
Mammalian Species ◽  
Sampling Site ◽  
Water Sources ◽  
Sequence Diversity ◽  
Rt Pcr ◽  
Reverse Transcription Pcr

ABSTRACT Reoviruses are a common class of enteric viruses capable of infecting a broad range of mammalian species, typically with low pathogenicity. Previous studies have shown that reoviruses are common in raw water sources and are often found along with other animal viruses. This suggests that in addition to the commonly monitored enteroviruses, reoviruses might serve as an informative target for monitoring fecal contamination of drinking water sources. Mammalian reoviruses were detected and identified by a combined cell culture–reverse transcription-PCR (RT-PCR) assay with novel primers targeting the L3 gene that encodes the λ3 major core protein. Five of 26 (19.2%) cytopathic effect-positive cell culture lysates inoculated with surface water were positive for reoviruses by RT-PCR. DNA sequence analysis of RT-PCR products revealed significant sequence diversity among isolates, which is consistent with the sequence diversity among previously characterized mammalian reoviruses. Sequence analysis revealed persistence of a reovirus genotype at a single sampling site, while a sample from another site contained two different reovirus genotypes.

Phylogeny and taxonomy of the family Arthrodermataceae (dermatophytes) using sequence analysis of the ribosomal ITS region

1999 ◽  
Vol 37 (2) ◽  
pp. 105-114 ◽  
Author(s):  
Y. GRAser ◽  
M. EL Fari ◽  
R. Vilgalys ◽  
A. F. A. Kuijpers ◽  
G. S. DE Hoog ◽  
...  
Keyword(s):  
Sequence Analysis ◽  
Its Region ◽  
The Family

scholarly journals Extensive Phylogenetic Analysis of Piscine Orthoreovirus Genomic Sequences Shows the Robustness of Subgenotype Classification

Pathogens ◽  
2021 ◽  
Vol 10 (1) ◽  
pp. 41
Author(s):  
Marcos Godoy ◽  
Daniel A. Medina ◽  
Rudy Suarez ◽  
Sandro Valenzuela ◽  
Jaime Romero ◽  
...  
Keyword(s):  
Phylogenetic Analysis ◽  
Genetic Variation ◽  
Sequence Analysis ◽  
Molecular Phylogeny ◽  
Phylogenetic Trees ◽  
Genomic Segment ◽  
Double Stranded Rna ◽  
The Family ◽  
Original Classification

Piscine orthoreovirus (PRV) belongs to the family Reoviridae and has been described mainly in association with salmonid infections. The genome of PRV consists of about 23,600 bp, with 10 segments of double-stranded RNA, classified as small (S1 to S4), medium (M1, M2 and M3) and large (L1, L2 and L3); these range approximately from 1000 bp (segment S4) to 4000 bp (segment L1). How the genetic variation among PRV strains affects the virulence for salmonids is still poorly understood. The aim of this study was to describe the molecular phylogeny of PRV based on an extensive sequence analysis of the S1 and M2 segments of PRV available in the GenBank database to date (May 2020). The analysis was extended to include new PRV sequences for S1 and M2 segments. In addition, subgenotype classifications were assigned to previously published unclassified sequences. It was concluded that the phylogenetic trees are consistent with the original classification using the PRV genomic segment S1, which differentiates PRV into two major genotypes, I and II, and each of these into two subgenotypes, designated as Ia and Ib, and IIa and IIb, respectively. Moreover, some clusters of country- and host-specific PRV subgenotypes were observed in the subset of sequences used. This work strengthens the subgenotype classification of PRV based on the S1 segment and can be used to enhance research on the virulence of PRV.

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