Quasispecies composition and phylogenetic analysis of feline coronaviruses (FCoVs) in naturally infected cats

Battilani, Mara; Coradin, Tiziana; Scagliarini, Alessandra; Ciulli, Sara; Ostanello, Fabio; Prosperi, Santino; Morganti, L.

doi:10.1016/s0928-8244(03)00237-2

Cited by 31 publications

(44 citation statements)

References 26 publications

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“…However, even in the context of molecular analysis, it is essential to make an informed choice among the available methods (e.g. in the choice of the target region and/or primers) because of rapid viral evolution, typical of RNA viruses, as it introduces variables in the gene sequence that can affect the interpretations of the results depending on the method applied (Battilani et al., ; Konig et al., ; Ciulli et al., ; Luzzago et al., ).…”

Section: Discussionmentioning

confidence: 99%

Evidence for Tunisian-Like Pestiviruses Presence in Small Ruminants in Italy Since 2007

Ciulli

Purpari

Agnello

et al. 2016

Transbound Emerg Dis

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The genus Pestivirus, which belongs to the Flaviviridae family, includes ssRNA+ viruses responsible for infectious diseases in pigs, cattle, sheep, goats and other domestic and wild ruminants. Like most of the RNA viruses, pestivirus has high genome variability with practical consequences on disease epidemiology, diagnosis and control. In addition to the officially recognized species in the genus Pestivirus, such as BVDV-1, BVDV-2, BDV and CSFV, other pestiviruses have been detected. Furthermore, most of the ruminant pestiviruses show low or absent species specificity observed in serological tests and are able to infect multiple species. Particularly, small ruminants are receptive hosts of the most heterogeneous group of pestiviruses. The aim of this study was to carry out the molecular characterization of pestiviruses isolated from sheep and goats in Sicily, Italy. Phylogenetic analysis of two viral genomic regions (a fragment of 5'-UTR and the whole N regions) revealed the presence of different pestivirus genotypes in the analysed goat and sheep herds. Two of five viral isolates were clustered with BVDV-1d viruses, a strain widespread in Italy, but never reported in Sicily. The other three isolates formed a distinct cluster with high similarity to Tunisian isolates, recently proposed as a new pestivirus species. This represents the first evidence for Tunisian-like pestivirus presence in small ruminants in Italy. Furthermore, one of the isolates was collected from a goat, representing the first isolation of Tunisian-like pestivirus from this species.

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Section: Discussionmentioning

confidence: 99%

Evidence for Tunisian-Like Pestiviruses Presence in Small Ruminants in Italy Since 2007

Ciulli

Purpari

Agnello

et al. 2016

Transbound Emerg Dis

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“…The coexistence of different variants of FIPV in diseased cat tissues has been reported by others using genomic techniques, including denaturating gel electrophoresis on amplicons from spike (S) and N genes, 11 and single-strand conformational polymorphism on N gene and open reading frame 7b amplicons. 3,23 It is worth noting that coronavirus N protein conservation is known to be low. The present study took a different approach, demonstrating heterogeneity at a viral protein level, namely the N protein, with IHC.…”

Section: Discussionmentioning

confidence: 99%

“…detect detail distribution of viral genome because they rely on homogenates. 3,11,23 It should be kept in mind that other processes, different from genomic changes, may be involved in N protein epitope alteration, namely alternative splicing and post-translational protein modifications. Alternative splicing as a cause of epitope change in FCoV could be excluded because plus-strand RNA synthesis sensitivity to ultraviolet light was found to be proportional to RNA length, ruling out splicing in subgenomic RNA formation in coronavirus species, namely, mouse hepatitis and infectious bronchitis coronaviruses.…”

Section: Discussionmentioning

confidence: 99%

“…12,21,34 Reverse transcriptase polymerase chain reaction (RT-PCR) amplification and product sequencing on RNA extracted from cat feces or rectal swabs have shown the emergence of genetic variants in the FCoV (FECV biotype) when sequential samples from the same cat are examined; similar results were obtained using samples from different cats within the same animal colony. 2,3,13,14 Such genetic drift seemingly takes place in the lower intestine, where FECV multiplies. 14 The DNA sequence variations between FECV and FIPV in samples obtained from a single cattery are less diverse than those found when the 2 viral biotypes are compared from different catteries.…”

Section: Introductionmentioning

confidence: 99%

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Detection of Antigenic Heterogeneity in Feline Coronavirus Nucleocapsid in Feline Pyogranulomatous Meningoencephalitis

Poncelet¹,

Coppens²,

Peeters

et al. 2008

Vet Pathol

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Abstract.A new monoclonal antibody (mAb), CCV2-2, was compared with the widely used FIPV3-70 mAb, both directed against canine coronavirus (CCoV), as a diagnostic and research tool. Western blot showed that both anti-CCoV mAbs only reacted with a protein of 50 kD, a weight consistent with the feline coronavirus (FCoV) viral nucleocapsid. A competitive inhibition enzyme-linked immunosorbent assay showed that the 2 recognized epitopes are distinct. Preincubation of CCV2-2 mAb with FCoV antigen suppressed the immunostaining. Formalin-fixed, paraffin-embedded sections from brains of 15 cats with the dry form of feline infectious peritonitis (FIP) were examined by immunohistochemistry. Immunohistochemistry was performed with both anti-CCoV mAbs, either on consecutive or on the same sections. A myeloid-histiocytic marker, MAC 387, was also used to identify FIP virus-infected cells. In all regions where MAC 387-positive cells were present, positive staining with the CCV2-2 mAb was systematically detected, except at some levels in 1 cat. In contrast, none or only a few cells were positive for the FIPV3-70 mAb. Double immunostaining showed macrophages that were immunopositive for either CCV2-2 alone or alternatively for CCV2-2 and FIPV3-70 mAbs. This reveals the coexistence of 2 cohorts of phagocytes whose FIP viral contents differed by the presence or absence of the FIPV3-70-recognized epitope. These findings provide evidence for antigenic heterogeneity in coronavirus nucleocapsid protein in FIP lesions, a result that is in line with molecular observations. In addition, we provide for the first time morphologic depiction of viral variants distribution in these lesions.

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“…The most precise method to detect viral heterogeneity is cloning and sequencing of a sufficient number of clones [3]. Because this is a laborious and expensive method, other mutation scanning approaches, such as SSCP, are often used as an alternative method for the screening of the quasispecies [4][5][6].…”

Section: Introductionmentioning

confidence: 99%

Comparative analysis of CE‐SSCP to standard RFLP‐CE‐FLA method in quantification of known viral variants within an RNA virus quasispecies

et al. 2011

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RNA viruses display the highest replication error rate in our biosphere, leading to highly diverse viral populations termed quasispecies. The gold standard method for detection and quantification of variants in a quasispecies is cloning and sequencing, but it is expensive, laborious and time consuming. Therefore, other mutation detection approaches, including SSCP, are often used. In this study, we demonstrate development and the usage of a CE-SSCP method for quantification of two nearly identical viral variants in heterogenic population of a mumps virus strain and its comparison to RFLP-CE-fragment length analysis (RFLP-CE-FLA). Analyzed PCR fragments were of the same size (245 bp) with one difference in their nucleotide sequence. The limit of detection of both methods was at 5% of the minor variant. When PCR amplicons of the two variants were pooled, methods' results were very similar. On the contrary, the quantification results of samples in which variants were mixed prior to PCR showed substantial difference between the two methods. Our results indicate that although both methods can be used for detection and monitoring of a specific mutation within a viral population, caution should be taken when quantitative analysis of complex samples is based solely on results of one method.

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Quasispecies composition and phylogenetic analysis of feline coronaviruses (FCoVs) in naturally infected cats

Cited by 31 publications

References 26 publications

Evidence for Tunisian-Like Pestiviruses Presence in Small Ruminants in Italy Since 2007

Evidence for Tunisian-Like Pestiviruses Presence in Small Ruminants in Italy Since 2007

Detection of Antigenic Heterogeneity in Feline Coronavirus Nucleocapsid in Feline Pyogranulomatous Meningoencephalitis

Comparative analysis of CE‐SSCP to standard RFLP‐CE‐FLA method in quantification of known viral variants within an RNA virus quasispecies

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