Strategies for the sequence determination of viral dsRNA genomes

Attoui, Houssam; Billoir, Frédérique; Cantaloube, Jean-François; Biagini, Philippe; Micco, Philippe de; Lamballerie, Xavier de

doi:10.1016/s0166-0934(00)00212-3

Cited by 172 publications

(128 citation statements)

References 9 publications

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“…Extraction and purification of dsRNA and synthesis of randomly primed cDNA were carried out as described (23,37). Primers (primer sequences will be supplied upon request) designed by using PulV and NBV small genome segment sequences were used for PCR amplification and sequencing of the MelV small genome segments.…”

Section: Methodsmentioning

confidence: 99%

“…Primers (primer sequences will be supplied upon request) designed by using PulV and NBV small genome segment sequences were used for PCR amplification and sequencing of the MelV small genome segments. Genome segment terminal sequences were obtained by using a two-step PCR amplification procedure, first by the single primer amplification technique (SPAT) (37), then by a seminested PCR using the combination of MelV genome segment-specific primers and the adaptor-specific primer used in the single primer amplification technique. Phylogenetic trees were constructed by using the neighbor-joining algorithm with bootstrap values determined by 1,000 replicates in the MEGA3 software package (38).…”

Section: Methodsmentioning

confidence: 99%

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A previously unknown reovirus of bat origin is associated with an acute respiratory disease in humans

Chua

Crameri

Hyatt

et al. 2007

Proc. Natl. Acad. Sci. U.S.A.

208

259

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Respiratory infections constitute the most widespread human infectious disease, and a substantial proportion of them are caused by unknown etiological agents. Reoviruses (respiratory enteric orphan viruses) were first isolated from humans in the early 1950s and so named because they were not associated with any known disease. Here, we report a previously unknown reovirus (named ''Melaka virus'') isolated from a 39-year-old male patient in Melaka, Malaysia, who was suffering from high fever and acute respiratory disease at the time of virus isolation. Two of his family members developed similar symptoms Ϸ1 week later and had serological evidence of infection with the same virus. Epidemiological tracing revealed that the family was exposed to a bat in the house Ϸ1 week before the onset of the father's clinical symptoms. Genome sequence analysis indicated a close genetic relationship between Melaka virus and Pulau virus, a reovirus isolated in 1999 from fruit bats in Tioman Island, Malaysia. Screening of sera collected from human volunteers on the island revealed that 14 of 109 (13%) were positive for both Pulau and Melaka viruses. This is the first report of an orthoreovirus in association with acute human respiratory diseases. Melaka virus is serologically not related to the different types of mammalian reoviruses that were known to infect humans asymptomatically. These data indicate that batborne reoviruses can be transmitted to and cause clinical diseases in humans.respiratory infection ͉ zoonosis ͉ human-to-human transmission ͉ orthoreovirus ͉ Pulau virus

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Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

A previously unknown reovirus of bat origin is associated with an acute respiratory disease in humans

Chua

Crameri

Hyatt

et al. 2007

Proc. Natl. Acad. Sci. U.S.A.

208

259

View full text Add to dashboard Cite

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“…cDNA copies of the dsRNA genome segments from HZ08 were synthesized, cloned, and sequenced according to a single-primer amplification technique described previously (2). The complete genome of HZ08 is 24,707 bp, and the size of the genome segments typically ranged from 1,027 to 3,927 bp.…”

mentioning

confidence: 99%

Complete Genome Sequence of a Reovirus Isolated from Grass Carp, Indicating Different Genotypes of GCRV in China

Wang

Zeng

Chun

et al. 2012

J Virol

151

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A widespread grass carp hemorrhagic disease (GCHD) caused by grass carp reovirus (GCRV) has been known in China since 1983. A virulent reovirus strain, HZ08, was isolated from diseased grass carp in Zhejiang Province, China. We sequenced and analyzed the complete genome of strain HZ08 and compared it with published GCRV genome sequences, contributing to the evidence of several genotypes of GCRV in China.

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“…Viral dsRNA from BN96/16 was prepared according to the protocol described by Attoui et al (1). Fulllength cDNA copies of each genomic segment were synthesized in a sequence-independent manner using the full-length amplification of cDNAs (FLAC) method, and subsequent rounds of PCR amplification were performed as described by Maan The data presented here are the first to report the complete sequence of the 10 genomic segments of a strain of BTV16.…”

mentioning

confidence: 99%

Complete Genomic Sequence of Bluetongue Virus Serotype 16 from China

Yang

Liu

et al. 2011

J Virol

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We report here the complete genomic sequence of the Chinese bluetongue virus serotype 16 (BTV16) strain BN96/16. This work is the first to document the complete genomic sequence (segments 1 to 10) of a BTV16 strain. The sequence information provided herein will help determine the geographic origin of BTV16 and define the phylogenetic relationship of BTV16 to other BTV strains.Bluetongue virus (BTV) is the "type" species of the genus Orbivirus, within the family Reoviridae (7). It is the etiological agent of bluetongue disease, which is listed as a "notifiable disease" by the Office International des Epizooties (OIE). The genome of BTV consists of 10 linear double-stranded RNA (dsRNA) segments encoding seven structural proteins (VP1 to VP7) and three nonstructural proteins (NS1, NS2, and NS3/NS3a) (5,6,8,9,11). Twenty-four BTV serotypes have thus far been recognized in the world, and a putative 25th serotype has recently been proposed (BTV25) (2). Seven BTV serotypes (BTV1, BTV2, BTV3, BTV4, BTV12, BTV15, and BTV16) have been isolated in China since the isolation of the first Chinese BTV strain in Yunnan Province in 1979 (3, 10, 12). Worldwide, there has been no report of the full genomic sequence of the BTV16 strain. It is necessary to acquire and analyze the sequence of the complete genome of a BTV16 strain to study the molecular features of this serotype.In the present study, we report the full genomic sequence of the serotype 16 BTV strain BN96/16, which was isolated from the blood of a sheep that showed obvious signs of bluetongue disease in Yunnan Province, China, in 1996. Viral dsRNA from BN96/16 was prepared according to the protocol described by Attoui et al. (1). Fulllength cDNA copies of each genomic segment were synthesized in a sequence-independent manner using the full-length amplification of cDNAs (FLAC) method, and subsequent rounds of PCR amplification were performed as described by Maan et al. (4). The 10 PCR products corresponding to each of the genomic segments of BN96/16 were purified and cloned into the pEASY-Blunt clone vector (Transgen Biotech, China). The cloned inserts were then sequenced by the Invitrogen Company to determine the full genomic sequence of BN96/16.The nucleotide sequences of the 10 BN96/16 segments were determined. The nucleotide lengths of BN96/16 segments 1 through 10 are as follows: segment 1, 3,944 bp; segment 2, 2,935 bp; segment 3, 2,772 bp; segment 4, 1,981 bp; segment 5, 1,763 bp; segment 6, 1,637 bp; segment 7, 1,156 bp; segment 8, 1,125 bp; segment 9, 1,052 bp; and segment 10, 822 bp. The amino acid (aa) lengths of the seven structural proteins of BN96/16 (VP1 to VP7) are as follows: VP1, 1,302 aa; VP2, 959 aa; VP3, 901 aa; VP4, 644 aa; VP5, 526 aa; VP6, 330 aa; and VP7, 349 aa. The amino acid lengths of the three nonstructural proteins of BN96/16 (NS1, NS2, and NS3/NS3a) are as follows: NS1, 552 aa; NS2, 354 aa; and NS3/NS3a, 229/216 aa.The data presented here are the first to report the complete sequence of the 10 genomic segments of a strain of BTV16. It is h...

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Strategies for the sequence determination of viral dsRNA genomes

Cited by 172 publications

References 9 publications

A previously unknown reovirus of bat origin is associated with an acute respiratory disease in humans

A previously unknown reovirus of bat origin is associated with an acute respiratory disease in humans

Complete Genome Sequence of a Reovirus Isolated from Grass Carp, Indicating Different Genotypes of GCRV in China

Complete Genomic Sequence of Bluetongue Virus Serotype 16 from China

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