Purification and characterization of the infectious hypodermal and haematopoietic necrosis virus of penaeid shrimps

Bonami, Jean-Robert; Trumper, Bronwen; Mari, Jocelyne; Brehélin, Michel; Lightner, Donald V.

doi:10.1099/0022-1317-71-11-2657

Cited by 136 publications

(88 citation statements)

References 20 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…IHHNV is a small, icosahedral, non-enveloped virus containing a single-stranded linear DNA genome approximately 4.1 kb in length (Bonami et al 1990, Mari et al 1993. Based on size, morphology and biochemical structure, IHHNV is considered to be a member of the family Parvoviridae (Bonami et al 1990).…”

Section: Introductionmentioning

confidence: 99%

Detection and quantification of infectious hypodermal and hematopoietic necrosis virus in penaeid shrimp by real-time PCR

Tang¹,

Lightner²

2001

Dis. Aquat. Org.

View full text Add to dashboard Cite

A real-time PCR method using a fluorogenic 5' nuclease assay and a PE Applied Biosystems GeneAmp 5700 sequence detector was developed to detect infectious hypodermal and hematopoietic necrosis virus (IHHNV) in penaeid shrimp. A pair of PCR primers to amplify an 81 bp DNA fragment and a fluorogenic probe (TaqMan probe) were selected from ORF1 (open reading frame 1) of the IHHNV genome. The primers and TaqMan probe used in this assay were shown to be specific for IHHNV and did not react with either hepatopancreatic parvovirus (HPV), white-spot syndrome virus (WSSV), or shrimp DNA. A plasmid, pIHHNV-P4, containing the target IHHNV sequence was constructed and used as a positive control. The concentration of pIHHNV-P4 was determined through spectrophotometric analysis and the plasmid was used for quantitative studies. This real-time PCR assay had a detection limit of 10 copies and a log-linear range up to 5 × 10 7 copies of IHHNV DNA. The assay was then used to quantify IHHNV in infected shrimp collected from 5 locations: Hawaii, Panama, Mexico, Guam, and the Philippines. The quantitative analysis showed that wild-caught, large juvenile Penaeus stylirostris collected from the Gulf of California (Mexico) in 1996 were naturally infected with IHHNV and contained up to 10 9 copies of IHHNV µg -1 of DNA. Similar quantities of IHHNV were detected in hatchery-raised, small juvenile P. stylirostris collected from Guam in 1995 and in farm-raised, post-larval P. monodon from the Philippines in 1996. Laboratory-infected P. stylirostris contained approximately 10 8 copies of IHHNV 31 d after being fed with IHHNV-infected shrimp tissue. In contrast, individuals of Super Shrimp ® , a line of P. stylirostris selected for IHHNV resistance, showed no signs of infection 32 d after ingesting IHHNV-infected shrimp tissue. Laboratory-infected P. vannamei also contained approximately 10 8 copies of IHHNV 30 d after being fed infected shrimp tissue. A time-course study of IHHNV replication in juvenile P. vannamei showed that the doubling time in the exponential growth phase was approximately 22 h.

show abstract

Section: Introductionmentioning

confidence: 99%

Detection and quantification of infectious hypodermal and hematopoietic necrosis virus in penaeid shrimp by real-time PCR

Tang¹,

Lightner²

2001

Dis. Aquat. Org.

View full text Add to dashboard Cite

show abstract

“…The average buoyant density of this virus by CsCl density gradient centrifugation is 1.40 g/ml [9]. By SDS-PAGE analysis, it was estimated that the purified virions contains four polypeptides of 74, 47, 39, and 37.5 kDa [9]. Recently Hou et al [31] reported the selfassembly of recombinantly expressed IHHNV capsid protein (CP) into virus-like particles (VLP's) in its size and shape and IHHNV-VLP's to encapsidate RNA and DNA of 0.5 kb size.…”

Section: Morphologymentioning

confidence: 99%

“…The multiple alignment of amino acid sequences of ORF3 of the Indian IHHNV isolate shows 11 amino acid substitutions compared to its closest relatives (Taiwan; AY355307 and Thailand; AY102034) [69]. Clearly, there is a discrepancy between the number of polypeptides visualized by SDS-PAGE at 4 [9] and the number of ORFs at 3 [69]. SDS-PAGE sizes compared to molecular weight are very similar; NS1 at 74 vs 75.77; NS2 at 47 and 39 vs 42.11 and VP1 at 37.5 vs 37.48 kDa respectively.…”

Section: Genome Organizationmentioning

confidence: 99%

“…Based on physicochemical and ultrastructural properties of the virion and genomic organization, IHHNV is taxonomically placed under the Parvoviridae family [9,55,74]. These are small icosahedral, nonenveloped viral particles 20-22 nm in diameter containing a single-stranded, linear DNA of approximately 3.9 kb [44,69].…”

Section: Morphologymentioning

confidence: 99%

“…These are small icosahedral, nonenveloped viral particles 20-22 nm in diameter containing a single-stranded, linear DNA of approximately 3.9 kb [44,69]. The average buoyant density of this virus by CsCl density gradient centrifugation is 1.40 g/ml [9]. By SDS-PAGE analysis, it was estimated that the purified virions contains four polypeptides of 74, 47, 39, and 37.5 kDa [9].…”

Section: Morphologymentioning

confidence: 99%

See 2 more Smart Citations

Genomics, Molecular Epidemiology and Diagnostics of Infectious hypodermal and hematopoietic necrosis virus

et al. 2012

View full text Add to dashboard Cite

Infectious hypodermal and hematopoietic necrosis virus (IHHNV) is one of the major viral pathogens of penaeid shrimps worldwide, which has resulted in severe mortalities of up to 90 % in cultured Penaeus (Litopenaeus) stylirostris from Hawaii and hence designated Penaeus stylirostris densovirus (PstDNV). IHHNV is distributed in shrimp culture facilities worldwide. It causes large economic loss to the shrimp farming industry. Our knowledge about the natural reservoirs of IHHNV is still scarce. Recent studies suggest that there is sufficient sequence variation among the isolates from different locations in Asia, suggesting multiple geographical strains of the virus. Four complete genomes and several partial sequences of the virus are available in the GenBank. Complete genome information would be useful for assessing the specificity of diagnostics for viruses from different geographical areas. Comparisons of complete genome sequences will help us gain insights into point mutations that can affect virulence of the virus. In addition, because of unavailability of shrimp cell lines for culturing IHHNV in vitro, quantification of virus is difficult. The recent progress in research regarding clinical signs, geographical distribution, complete genome sequence and genetic variation, transmission has made it possible to obtain information on IHHNV. A comprehensive understanding of IHHNV infection process, pathogenesis, structural proteins and replication is essential for developing prevention measures. To date, no effective prophylactic measure for IHHNV infection is available for shrimp to reduce its impact. This review provides an overview of key issues regarding IHHNV infection and disease in commercially important shrimp species.

show abstract

Phylogenetic and recombination analysis of genomic sequences of IHHNV

Shen

Zhang

Shao

2015

J. Basic Microbiol.

View full text Add to dashboard Cite

Infectious hypodermal and hematopoietic necrosis virus (IHHNV) is one of the most recurrent viral pathogens infecting penaeid shrimps. Although previous reports showed that genomic recombination happened among different IHHNV strains, no study has been carried out to systematically elucidate such recombination. In this study, we performed the phylogenetic and recombination analysis of IHHNV the available complete genomes sequences in GenBank. We have found two intra-group recombination between lineages III and II of infectious group, one inter-lineage recombination between lineage II of infectious group and non-infectious group among IHHNV strains.

show abstract

Purification and characterization of the infectious hypodermal and haematopoietic necrosis virus of penaeid shrimps

Cited by 136 publications

References 20 publications

Detection and quantification of infectious hypodermal and hematopoietic necrosis virus in penaeid shrimp by real-time PCR

Detection and quantification of infectious hypodermal and hematopoietic necrosis virus in penaeid shrimp by real-time PCR

Genomics, Molecular Epidemiology and Diagnostics of Infectious hypodermal and hematopoietic necrosis virus

Phylogenetic and recombination analysis of genomic sequences of IHHNV

Contact Info

Product

Resources

About