In situ Detection of Reovirus in Formalin-Fixed, Paraffin-Embedded Chicken Tissues Using a Digoxigenin-Labeled cDNA Probe

Liu, Hung-Jen; Giambrone, J. J.

doi:10.2307/1592203

Cited by 14 publications

(4 citation statements)

References 16 publications

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“…The use of this technique has become more attractive with the availability of safer, more stable, nonradioactive probes (Azzi et al, 1990) and ISH has been developed for the detection of various viral pathogens of poultry (Hathcook & Giambrone, 1992;Nielsen et al, 1995;Liu & Giambrone, 1997). Although ISH has been used to identify several mammalian retroviruses, the use of this sensitive tool for the detection of avian retroviruses has not been reported.…”

Section: Introductionmentioning

confidence: 99%

Tropism of subgroup J avian leukosis virus as detected byin situhybridization

et al. 1999

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The HPRS-103 strain of avian retrovirus is the prototype of subgroup J avian leukosis virus (ALV-J) and causes myeloid leukosis in meat-type chickens. Using immunohistochemical detection of the viral groupspecific antigen (Gag) we have previously demonstrated that the induction of myeloid leukosis by ALV-J is associated with viral tropism for myelomonocytic cells. In this paper we describe an in situ hybridization (ISH) technique using digoxigenin (DIG)-labelled probes for detecting RNA transcripts in tissues from chickens infected with avian leukosis viruses (ALV) of subgroups J (HPRS-103 strain) and A (RAV-1 strain). Virus-specific RNA was detected mainly in the heart, kidney, proventriculus and adrenal in locations similar to those of the Gag protein. Viral gene expression could not be detected in the bone marrow or tumour tissues using this test. Higher levels of viral gene expression in the bursa of Fabricius infected with RAV-1, but not with HPRS-103, might help explain the inability of the latter virus to induce lymphoid leukosis.

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Section: Introductionmentioning

confidence: 99%

Tropism of subgroup J avian leukosis virus as detected byin situhybridization

et al. 1999

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“…It plays an important role in ARV pathogenesis due to its capability to elicit a neutralization antibody reaction in virus infection (Wickramasinghe et al, 1993). Both classical and molecular-based techniques have been developed for detection of ARV (Li et al, 1996;Liu & Giambrone, 1997;Liu et al, 1999aLiu et al, ,b, 2000Liu et al, , 2004Ke et al, 2006). Virus isolation is a reliable diagnostic method but not suitable for rapid diagnosis.…”

Section: Discussionmentioning

confidence: 99%

“…Several diagnostic techniques based on antigen and antibody reactions have been developed (Slaght et al, 1978;Islam & Jones, 1988;Lee et al, 1994;Li et al, 1996;Liu et al, 1999aLiu et al, ,b, 2000Liu et al, , 2002Liu et al, , 2004Liu & Giambrone, 1997;Chen et al, 2004;Ke et al, 2006). In this study, eight monoclonal antibodies (mAbs) against sC and three mAbs against His were produced.…”

Section: Introductionmentioning

confidence: 99%

Development and characterization of monoclonal antibodies against avian reovirus σC protein and their application in detection of avian reovirus isolates

et al. 2006

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Avian reovirus (ARV) is a non-enveloped virus with a segmented double-stranded RNA genome surrounded by a double icosahedral capsid shell. ARVs are associated with viral arthritis, immunosuppression, and enteric diseases in poultry. The sC protein was involved in induction of apoptosis and neutralization antibody. In the present study, sC-His protein was expressed in Sf9 insect cells and purified by immobilized metal affinity chromatography. Eight monoclonal antibodies (mAbs) against sC-His and three mAbs against His were screened from hybridoma cells produced by fusion of splenocytes from immunized mice with NS1 myeloma cells. Among the eight mAbs against sC protein, all belonged to the IgG isotype except three for IgM. It was discovered that all anti-His mAbs were mixtures of IgG and IgM isotypes. mAbs reacted with sC-His protein in a conformation-independent manner based on dot blot and western blotting assays. The competitive binding assay indicated that all mAbs recognized the same epitope on sC protein that was conserved in different isolates. Compared with the commercial anti-ARV S1133 polyclonal antibody, mAb (D15) had universal reactivity to all serotypes or genotypes of ARVs tested. This monoclonal antibody may therefore be useful for the development of an antigen-capture enzyme-linked immunosorbent assay for rapid detection of field isolates.

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“…acts as apoptosis inducer [7]. Thenucleotide (nt) homology and aa homology of σC have been found suitable for comparison among different strains; firstly, the σC protein is the most variable protein in the ARV [8,9] for both very hypervariable aa regions one to 122 and 196 to 326 [10]; and secondly, it induces the production of neutralizing antibodies [11].…”

Section: Veterinary Science and Medicinementioning

confidence: 99%

The σC Gene Characterization of Seven Turkey Arthritis Reovirus Field Isolates in Pennsylvania during 2011-2014

2015

J Vet Sci Med

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Seven turkey arthritis reovirus (TARV) field isolates made in our laboratory during 2011-2014 were characterized by sequencing of reovirus σC genes. Of the seven TARV field isolates, six were isolated from 3-to-17-week-old turkeys with tenosynovitis in six Pennsylvania farms and one was isolated from an Indiana turkey case submitted to our lab in 2014. This report describes the amplification and sequencing of the σC genes for genetic characterization studies on the seven TARV field isolates. Phylogenetic analysis of the sequence data of the seven TARV isolates together with recent published five TARVs detected in Minnesota (MN) and 25 avian reovirus (ARV) strains retrieved from GenBank revealed that all the seven PA TARV field isolates and five MN TARVs fit into genotyping cluster two when compared with a total of five different genotyping clusters (cluster 1-5) generated by the TARV and ARV reference strains. Comparison of amino acidsequences of the seven TARV isolates in cluster two with ARV vaccine strains (S1133, 1733, and 2048) in cluster one revealed that there were less than 60% similarity in nucleotide sequence and less than 56% in amino acid sequence between the two clusters. However the seven PA TARV isolates shared greater than 99% similarity with each other. Our research findings have indicated that the seven PA TARV field isolates and the five MN TARVs are grouped in the same genotype two, a separate genotype or virus species within the Orthoreovirus genus.

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In situ Detection of Reovirus in Formalin-Fixed, Paraffin-Embedded Chicken Tissues Using a Digoxigenin-Labeled cDNA Probe

Cited by 14 publications

References 16 publications

Tropism of subgroup J avian leukosis virus as detected byin situhybridization

Tropism of subgroup J avian leukosis virus as detected byin situhybridization

Development and characterization of monoclonal antibodies against avian reovirus σC protein and their application in detection of avian reovirus isolates

The σC Gene Characterization of Seven Turkey Arthritis Reovirus Field Isolates in Pennsylvania during 2011-2014

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