Sequence of a cDNA carrying the glycoprotein gene and part of the matrix protein M2 gene of viral haemorrhagic scepticaemia virus, a fish rhabdovirus

Thiry, Michel; Lecoq-Xhonneux, F.; Dheur, I.; Renard, A.; Kinkelin, P. de

doi:10.1016/0167-4781(91)90200-6

Cited by 41 publications

(33 citation statements)

References 8 publications

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“…"Numbering of nucleotides according to the open readlng frame (start codon ATG) b~c c o r d l n g to Thiry et al (1991) CAccording to Koener et al (1987) The purified RNA was usually eluted in 30 to 100 1 1 RNase-free water. The quality and the quantlty of the purified nucleic acids were determined by measuring the absorbance at 260 nm (A,,,,) and 280 nm (A280) in a spectrophotometer (Zeiss, M4QIII + PMQII + H30DS).…”

Section: Methodsmentioning

confidence: 99%

“…They used primers specific for the N-gene. Cloning and sequencing of cDNAs of IHNV and VHSV glycoprotein genes (Koener et al 1987, Thiry et al 1991 revealed considerable differences at the nucleotide as well as at the amino acid level. To distingu~sh IHNV and VHSV by RT-PCR, Bruchhof et al (1995) used primer pairs designed for the amplification of glycoprotein G-specific gene fragments of the 2 viruses.…”

Section: Introductionmentioning

confidence: 99%

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Rapid and sensitive reverse transcriptase-polymerase chain reaction based detection and differential diagnosis of fish pathogenic rhabdoviruses in organ samples and cultured cells

Miller¹,

Rapp²,

U³

et al. 1998

Dis. Aquat. Org.

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A reverse transcnptase-polymerase chain reaction (RT-PCR) assay was developed and applied to the detection and differentiation of viral haemorrhagic septicaemia virus (VHSV) and infectious hematopoietic necrosis virus (IHNV) in organ samples and cultured cells, regardless of the serotype. This method was developed by selecting primer sets corresponding to highly conserved regions of the glycoprotein G-gene sequences of the 2 vlruses. The very fast RNA extraction, reverse transcnption and PCR permitted us to read the agarose gels within 7 to 9 h after samples, cultured cells and whole fish arrived, which is of great importance when there is reason to believe that VHSV or IHNV may be present This is also the first report of a large-scale field trial companng the RT-PCR assay in trout from 30 German fish farms (a total of 330 rainbow trout) with the usual virus isolation and identification method in order to evaluate the efficiency of the RT-PCR assay for general use in fish health management programs. RT-PCR followed by semi-nested PCR uslng RNA dlrectly extracted from fish tissue turned out to be the most sensitive method. It recognized 9 fish farms a s VHS-positive and 7 as IHN-positive. This is 3 VHS-and 4 IHN-farms more than detected by the traditional virus isolation method. By directly examining the tissue by means of a PCR test ~t was possible to detect viral RNA in acutely and subacutely to chronically diseased fish as well as in asyrnptomatlc VHS/IHNcarner fish Therefore, this effectlve and powerful assay for detecting VHSV and IHNV by means of PCR has great advantages compared with the presently used procedures KEY WORDS: RT-PCR . VHSV/IHNV . Differential diagnosis in organ samples INTRODUCTIONAquatic rhabdoviruses such as viral haemorrhagic septicaemia virus (VHSV) and infectious hematopoietic necrosis virus (IHNV) are significant pathogens for salmonids. VHSV and IHNV often causes diseases with high mortality among salmonids, especially in rainbow trout Oncorhynchus mykiss.Since its first isolation (Jensen 1965) VHSV has been widely detected in Europe. VHSV was first isolated in 1988 in chinook salmon Oncorhynchus tshawytscha and in coho salmon 0. kisutch on the West coast of North America (Brunson et al. 1989, Hopper 1989. Virus isolations from Pacific cod Gadus macrocephalus and Alaskan Pacific herring Clupea harengus pallasi were reported in 1992 and 1993, respectively (Meyers et al. 1992(Meyers et al. , 1993. It was concluded that VHSV had been enzootic in the North Pacific region for some time (Batts et al. 1993). Even in the British Isles, thought to be VHSV-free, VHSV was isolated from a turbot Scophthalmus maximus farm in Scotland (Ross et al. 1994). Experimental infection showed that this turbot isolate was of low virulence for rainbow trout 0 Inter-Research 1998 Resale of full article not permitted

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Section: Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Rapid and sensitive reverse transcriptase-polymerase chain reaction based detection and differential diagnosis of fish pathogenic rhabdoviruses in organ samples and cultured cells

Miller¹,

Rapp²,

U³

et al. 1998

Dis. Aquat. Org.

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“…It has a singlestranded RNA genome that encodes five structural proteins : the nucleocapsid protein N (Bernard et al, 1990), the polymerase-associated protein P, the matrix protein M (Benmansour et al, 1994), the transmembrane glycoprotein G (Thiry et al, 1991) and the polymerase L. In addition, a sixth Author for correspondence : Abdenour Benmansour.…”

Section: Introductionmentioning

confidence: 99%

Mutations in the glycoprotein of viral haemorrhagic septicaemia virus that affect virulence for fish and the pH threshold for membrane fusion.

Gaudin

Kinkelin

Benmansour

1999

Journal of General Virology

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To study the molecular basis of virulence of viral haemorrhagic septicaemia virus (VHSV), we used a cross-reactive neutralizing MAb to select MAb-resistant (MAR) mutants with reduced pathogenicity for fish. From sequence determination of the G gene of MAR mutants, attenuated laboratory variant and avirulent field strains, we identified two distant regions of the glycoprotein associated with virulence : region I (aa 135-161), homologous to the putative fusion peptide of both rabies virus (RV) and vesicular stomatitis virus (VSV), and region II (surrounding aa 431-433), homologous to RV and VSV domains controlling the conformational changes necessary for the fusion process to take place. Simultaneous mutations in both regions resulted in the most attenuated phenotype and we obtained genetic evidence that regions I and II may be structurally linked. As the MAR mutants had mutations in or near domains involved in fusion, the fusion properties of VHSV and its variants were analysed. This work allowed us to postulate that the fusion domain of VHSV is probably constituted of two distinct regions of the protein connected through a disulfide bridge between cysteines 110 and 152. Finally, we obtained evidence suggesting that the pH threshold for fusion is a determinant for virulence : restriction of fusion to a more acidic pH was associated with attenuation for the variant tr25 which had a shift of the threshold for maximal fusion from pH 6n30 (for the parental strain) to pH 6n00 ; conversely, two field strains which had maximal fusion at pH 6n60 were the most virulent.

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“…Oligonucleotide primers. The oligonucleotide primers were designed using the available sequences of the IPNV VP2, IHNV G or VHSV G genes (Koener et al 1987, Thiéry et al 1991, Pryde et al 1993. To ensure specificity, the primers were compared with the sequences deposited in the GenBank using FASTA software; the primers G4/G5 were used to amplify a 1512 bp segment of the IHN viral cDNA encoding the G protein; the G4 primer (5'-ATGATCACCACTCC-GCTCATT-3') was hybridized to Positions 1-21 (sense orientation) in the open reading frame (ORF) of the IHNV G gene; the G5 primer (5'-CCGGTTTGCC-AGGTGATACAT-3') was 21 nucleotides long and hybridized to Positions 1492-1512 of the ORF (antisense orientation).…”

mentioning

confidence: 99%

Comparison of two birnavirus-rhabdovirus coinfections in fish cell lines

Rodrı́guez¹,

Alonso²,

Pérez-Prieto³

2005

Dis. Aquat. Org.

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Aquabirnaviruses, such as the infectious pancreatic necrosis virus (IPNV), Novirhabdoviruses, such as the infectious hematopoiteic necrosis virus (IHNV) and the viral hemorrhagic septicemia virus (VHSV), cause considerable losses to the salmonid industry worldwide. Coinfections of 2 viruses have been described, but the interactions between rhabdoviruses and birnaviruses have not been examined closely. Using virus titration, flow cytometry and RT-PCR assays, we compared the effect of IPNV on the replication of IHNV and VHSV in tissue culture cells. RT-PCR assays indicated that simultaneous infection of IPNV with VHSV does not affect the replication of the rhabdovirus either in the first or successive passages; the infective titers were similar in single and double infections. In contrast, coinfection of IPNV with IHNV induced a fall in infectivity, with reduced expression of IHNV viral antigens in BF-2 cells from Lepomis macrochirus and a loss of 4.5 log 10 units of the infective titer after 3 successive passages. It was possible to stimulate BF-2 cells to produce significant interferon-like activity against IHNV but not against VHSV. KEY WORDS: Infectious pancreatic necrosis virus · Infectious hematopoiteic necrosis virus · Viral hemorrhagic septicemia virus · Coinfections · Fish viruses Resale or republication not permitted without written consent of the publisherDis Aquat Org 67: [183][184][185][186][187][188][189][190] 2005 provides some protection against the secondary ISAV infection (Johansen & Sommer 2001). Melby & Falk (1995) infected an IPNV carrier stock of Atlantic salmon with tissue homogenates containing the infectious salmon anemia virus (ISA), and demonstrated that ISA infection did not activate the latent IPNV infections. More recently, Pakingking et al. (2003) explored experimental dual infections in flounder Paralichthys olivaceus with an aquabirnavirus and the VHSV on the one hand, and bacteria on the other. They concluded that the primary infection suppresses the secondary viral infection, but facilitates the bacterial infection. Further, the studies by Brudesth et al. (2002) on interactions between VHSV and IHNV during an experimental coinfection of rainbow trout demonstrated a reduced systemic distribution of IHNV.All these reports indicate that interactions between viruses during simultaneous infections is as interesting a line of research in fishes, as it is proving to be in mammals. In previous studies we reported that IPNV interferes with the replication of IHNV in tissue culture cells and in fishes (Alonso et al. 1999a(Alonso et al. , 2003. The aim of the present study was to determine whether coinfections of VHSV or IHNV with IPNV present similar features; i.e. whether VHSV growth is affected in a similar way to that of IHNV and whether its infectivity is reduced. This is relevant to diagnoses, since the detection methods for viral coinfections would need to be improved, as rapid identification could prevent further spread. In addition, little is known about interacti...

show abstract

Sequence of a cDNA carrying the glycoprotein gene and part of the matrix protein M2 gene of viral haemorrhagic scepticaemia virus, a fish rhabdovirus

Cited by 41 publications

References 8 publications

Rapid and sensitive reverse transcriptase-polymerase chain reaction based detection and differential diagnosis of fish pathogenic rhabdoviruses in organ samples and cultured cells

Rapid and sensitive reverse transcriptase-polymerase chain reaction based detection and differential diagnosis of fish pathogenic rhabdoviruses in organ samples and cultured cells

Mutations in the glycoprotein of viral haemorrhagic septicaemia virus that affect virulence for fish and the pH threshold for membrane fusion.

Comparison of two birnavirus-rhabdovirus coinfections in fish cell lines

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