Localization and genotyping of canine papillomavirus in canine inverted papillomas

2021 ◽  
pp. 104063872110357
Author(s):  
Margherita Orlandi ◽  
Maurizio Mazzei ◽  
Marta Vascellari ◽  
Erica Melchiotti ◽  
Claudia Zanardello ◽  
...  
Keyword(s):  
In Situ Hybridization ◽  
Inclusion Bodies ◽  
Animal Species ◽  
Viral Etiology ◽  
Molecular Approaches ◽  
Dna And Rna ◽  
Promising Tool ◽  
First Choice ◽  
Inverted Papillomas

Numerous canine papillomaviruses (CPVs) have been identified (CPV1–23). CPV1, 2, and 6 have been associated with inverted papillomas (IPs). We retrieved 19 IPs from 3 histopathology archives, and evaluated and scored koilocytes, inclusion bodies, giant keratohyalin granules, cytoplasmic pallor, ballooning degeneration, and parakeratosis. IHC targeting major capsid proteins of PV was performed, and CPV genotyping was achieved by PCR testing. Tissue localization of CPV DNA and RNA was studied by chromogenic and RNAscope in situ hybridization (DNA-CISH, RNA-ISH, respectively). IPs were localized to the limbs (50%), trunk (30%), and head (20%), mainly as single nodules (16 of 19). In 15 of 19 cases, immunopositivity was detected within the nuclei in corneal and subcorneal epidermal layers. PCR revealed CPV1 in 11 IPs and CPV2 DNA in 3 IPs. Overall, 14 of 17 cases were positive by both DNA-CISH and RNA-ISH, in accord with PCR results. A histologic score >5 was always obtained in cases in which the viral etiology was demonstrated by IHC, DNA-CISH, and RNA-ISH. IHC and molecular approaches were useful to ascertain the viral etiology of IPs. Although IHC is the first choice for diagnostic purposes, ISH testing allows identification of PV type and the infection phase. RNA-ISH seems a promising tool to deepen our understanding of the pathogenesis of different PV types in animal species.

scholarly journals Rapid detection of Salmonella sp. in pork samples using fluorescent in situ hybridization: a comparison with VIDAS®-SLM system and ISO 6579 cultural method

2007 ◽  
Vol 59 (6) ◽  
pp. 1388-1393 ◽  
Author(s):  
M. Vieira-Pinto ◽  
M. Oliveira ◽  
F. Bernardo ◽  
C. Martins
Keyword(s):  
In Situ Hybridization ◽  
Rapid Detection ◽  
Fluorescent In Situ Hybridization ◽  
Diagnostic Assay ◽  
Promising Tool ◽  
Cultural Method ◽  
Inner Surface ◽  
Peptone Water ◽  
Pork Samples

This study reports the use of the fluorescent in situ hybridization (FISH) with Sal3 probe for Salmonella detection in swine carcasses inner surface (swab); and in the correspondent samples of ileum, ileocolic, and mandibular lymph nodes; and tonsils, after dilution (1:10) in buffered peptone water and a pre-enrichment step (37(0)C, 18h). In order to evaluate the efficiency of FISH, 235 naturally contaminated samples were simultaneously tested by the cultural method (ISO 6579) and by the Vitek Immuno Diagnostic Assay System (VIDAS®) - Salmonella (SLM) system. The cultural method identified 39 positive samples. From these, VIDAS®- SLM only detected 23. FISH identified 115 positive samples. This difference was highly significant (P<0.001). From positive samples, 32 were also confirmed by the cultural method. The results indicate FISH as a promising tool for rapid Salmonella detection in samples of pork and swine carcasses.

In Situ Hybridization: Detection of DNA and RNA

2003 ◽  
pp. 27-46 ◽  
Author(s):  
Long Jin ◽  
Xiang Qian ◽  
Ricardo V. Lloyd
Keyword(s):  
In Situ Hybridization ◽  
Dna And Rna ◽  
Hybridization Detection

A molecular cytogenetic comparison between Rhynchosciara americana and Rhynchosciara hollaenderi (Diptera: Sciaridae)

Genome ◽  
1993 ◽  
Vol 36 (5) ◽  
pp. 831-843 ◽  
Author(s):  
Ann Jacob Stocker ◽  
Eduardo Gorab ◽  
J. M. Amabis ◽  
F. J. S. Lara
Keyword(s):  
In Situ Hybridization ◽  
Species Complex ◽  
Polytene Chromosomes ◽  
Chromosomal Evolution ◽  
Molecular Cytogenetic ◽  
The Third ◽  
Dna And Rna ◽  
Rhynchosciara Americana ◽  
Telomeric Heterochromatin

The polytene chromosomes of Rhynchosciara americana and R. hollaenderi, a pair of sibling species in the americana-like group of Rhynchosciara, were compared using a number of techniques, including in situ hybridization. With classical cytological techniques, the only differences observed were in the morphology of centromeric and telomeric heterochromatin, in the size of a DNA and RNA puff, and in the presence of an inversion polymorphism in R. hollaenderi. However, after in situ hybridization with rDNA and poly-r(A) probes, differences between the two species appeared at a number of sites. Differences in poly-r(A) sites were especially informative in establishing phylogenetic relationships between these two species and a third species currently being examined from this group. Chromosomal evolution between these species appears to have occurred mainly through differential amplification and transposition of repetitive sequence DNA, of which dA:dT tracts are an important component. The R. hollaenderi karyotype is tentatively considered more ancestral than that of R. americana because it has features present in the third Rhynchosciara species. Explanations for the monomorphisms observed in Rhynchosciara species and mechanisms of speciation in the group are considered within the context of the species' complex behavior.Key words: Rhynchosciara, chromosome homology, in situ hybridization, phylogeny, evolution.

Localizing DNA and RNA within nuclei and chromosomes by fluorescence in situ hybridization

1991 ◽  
Vol 8 (2) ◽  
pp. 41-58 ◽  
Author(s):  
John A. McNeil ◽  
Carol Villnave Johnson ◽  
Kenneth C. Carter ◽  
Robert H. Singer ◽  
Jeanne Bentley Lawrence
Keyword(s):  
In Situ Hybridization ◽  
Fluorescence In Situ Hybridization ◽  
Dna And Rna

Lox10, a member of the NK-2 homeobox gene class, is expressed in a segmental pattern in the endoderm and in the cephalic nervous system of the leech Helobdella

Development ◽  
1993 ◽  
Vol 118 (3) ◽  
pp. 877-892 ◽  
Author(s):  
D. Nardelli-Haefliger ◽  
M. Shankland
Keyword(s):  
Central Nervous System ◽  
Nervous System ◽  
In Situ Hybridization ◽  
Animal Species ◽  
Homeobox Gene ◽  
Endoderm Differentiation ◽  
Homeodomain Sequence ◽  
Supraesophageal Ganglion ◽  
The Central Nervous System

A novel leech homeobox gene, Lox10, is shown to encode a homeodomain sequence characteristic of a phyletically widespread NK-2 homeobox gene class. Lox10 expression was examined in leech embryos of various ages by in situ hybridization. In the unsegmented cephalic region, Lox10 RNA is expressed in a subset of the cells descended from the a' and b' micromeres, including a small cluster of cells, believed to be postmitotic neurons, within the supraesophageal ganglion of the central nervous system. Hybridization signal was not detected in either the mesoderm or ectoderm of the trunk segments, and the apparent restriction of Lox10 ectodermal expression to the nonsegmented cephalic domain resembles the restricted forebrain expression pattern of its mammalian homologues. Lox10 is also expressed within the endodermal tissues of the leech midgut, which arises by cellularization from a polynucleate syncytium. Endodermal expression is organized into a pattern of transverse stripes and spots which are aligned with the intersegmental septa, and which prefigure the pattern of gut wall constrictions observed at later stages of development. Lox10 is the first molecular marker of segmentally periodic endoderm differentiation reported for any animal species.

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