Genetic variation and immunohistochemical differences among geographic isolates of Taura syndrome virus of penaeid shrimp

Robles‐Sikisaka, Refugio; Hasson, Kenneth W.; Garcia, Denise K.; Brovont, Katherine E.; Cleveland, Karyn D.; Klimpel, Kurt R.; Dhar, Arun K.

doi:10.1099/0022-1317-83-12-3123

Cited by 34 publications

(31 citation statements)

References 23 publications

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“…The introduction of P. vannamei culture in Taiwan, China, Thailand and Indonesia also resulted in the spread of TS virus (TSV) to these regions within a relatively short period of time (Tu et al 1999, Yu & Song 2000, Nielsen et al 2005. Several genotypes or strains are now known to exist and the virus remains an important infectious disease in countries where P. vannamei is farmed (Erickson et al 2002, Lien et al 2002, Robles-Sikisaka et al 2002, Rosenberry 2002.…”

Section: Introductionmentioning

confidence: 99%

Comparison of Taura syndrome virus (TSV) detection methods during chronic-phase infection in Penaeus vannamei

Poulos

Noble

Lightner³

2008

Dis. Aquat. Org.

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Methods to detect Taura syndrome virus (TSV) were assessed for their ability to detect the virus during chronic phase infection in the Pacific white shrimp Penaeus vannamei. In situ hybridization (ISH), immunohistochemistry (IHC) using monoclonal antibody 1A1, conventional RT-PCR and real-time quantitative (q)RT-PCR were compared using shrimp sampled over 60 wk following experimental TSV infection. Between Weeks 7 and 60, hematoxylin-eosin histology confirmed the presence of lymphoid organ spheroids (LOS) and an absence of lesions in the cuticular epithelium. ISH detected TSV in LOS over the duration of the study. IHC was generally less sensitive than ISH, and after Week 24, was often unable to confirm TSV infection. Detection of TSV by RT-PCR was highly dependent on sample source after Week 43, where viral RNA was detected in 12 of 14 hemolymph samples but only 5 of 16 pleopod samples. qRT-PCR detected TSV over the 60 wk in both hemolymph and pleopods, although RNA copy numbers in pleopods were consistently lower throughout the study. This study demonstrates that ISH and qRT-PCR are the most reliable methods for detecting TSV during late chronic phase infection. RT-PCR was also reliable if hemolymph was used as the sample source. KEY WORDS: Taura syndrome · TSV · RT-PCR · qRT-PCR · In situ hybridization · Immunohistochemistry Resale or republication not permitted without written consent of the publisherDis Aquat Org 82: [179][180][181][182][183][184][185] 2008 lasting ~5 d and a chronic phase that lingers in survivors of ecdysis. Shrimp with a chronic phase infection present no melanized lesions or abnormal behavior and histologically present normal cuticular epithelium. However, they display marked hypertrophy of the lymphoid organ (LO) with the presence of numerous LO spheroids (LOS) (Hasson et al. 1999b, Srisuvan et al. 2005.Over 48 wk following TSV infection of juvenile Penaeus vannamei, in situ hybridization (ISH) was able to detect TSV within the LOS up to Week 32, even though LOS remained evident thereafter up to Week 48 post-infection (pi) (Hasson et al. 1999b). RT-PCR and real-time quantitative (q)RT-PCR have been developed to detect TSV (Nunan et al. 1998) and the latter test was able to detect the virus in LO, gill and pleopod tissues in the chronic phase of the disease up to 23 wk pi when the study was terminated . A monoclonal antibody (1A1) generated to a TSV isolate from Hawaii (circa 1994) used for immunohistochemistry can detect the virus in fixed tissue sections but the reactivity is limited to related isolates (Poulos et al. 1999, Erickson et al. 2002. However, the utility of IHC has not been investigated during chronic phase infection.Studies to date have shown that accurate diagnosis of TSV is dependent upon the stage of infection and the nature of the isolate. The selection of diagnostic specimens and TSV detection methods becomes more important to diagnose the infection once disease signs abate as infection passes through the transition stage and the virus becomes sequestered...

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Section: Introductionmentioning

confidence: 99%

Comparison of Taura syndrome virus (TSV) detection methods during chronic-phase infection in Penaeus vannamei

Poulos

Noble

Lightner³

2008

Dis. Aquat. Org.

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“…In addition, all of the MAb reacted with multiple antigen bands in gels prepared from purified TSV (Poulos et al 1999). Application of one of the MAb (designated 1A1 and specific to VP1) for immunohistochemical detection of TSV infection in shrimp gave positive results with TSV isolates from Taiwan, USA and Hawaii, but negative results for TSV isolates from Mexico, Nicaragua and Belize (Robles-Sikisaka et al 2002, Erickson et al 2005. Phylogenetic analysis of the TSV capsid protein 2 gene (also called CP2 but equivalent to VP1) from various sources revealed 3 distinct geographic groups associated with Belize, the USA and Southeast Asia (Tang & Lightner 2005).…”

Section: Specificity Of Mabmentioning

confidence: 99%

“…MAb specific to VP1 (designated 1A1) was used to develop Western-blot, dot-blot and immunohistochemical assays for detection of various TSV strains. However, this MAb failed to detect several isolates of TSV from various sources (Erickson et al 2002, Robles-Sikisaka et al 2002. Polyclonal antibodies against recombinant proteins of VP1 and VP3 have also been developed (Chaivisuthangkura et al 2006).…”

Section: Abstract: Penaeus Vannamei · Monoclonal Antibody · Mab · Tamentioning

confidence: 99%

Specific monoclonal antibodies raised against Taura syndrome virus (TSV) capsid protein VP3 detect TSV in single and dual infections with white spot syndrome virus (WSSV)

Longyant

Poyoi²,

Chaivisuthangkura³

et al. 2008

Dis. Aquat. Org.

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“…ORF1 and ORF2 are predicted to encode the nonstructural and structural polyproteins, respectively. Phylogenic analysis based on the nucleotide sequences of CP2 suggested that there are 3 lineages, namely the Americas, Belize, and SE Asia, among the various geographic TSV isolates (Robles-Sikisaka et al 2002, Tang & Lightner 2005. In the CP1 and CP2 regions, low genetic variation (0 to 0.24% for CP1 and 0 to 0.35 or 0 to 5.6% for CP2 nucleotide sequences) was observed among the isolates analyzed.…”

Section: Introductionmentioning

confidence: 95%

“…TSV disease has spread widely via infected broodstock throughout America and Southeast Asia, causing severe economic losses in the shrimp aquaculture industry in the last decade (Brock 1997, Tu et al 1999, Robles-Sikisaka et al 2002, Chang et al 2004. TSV is known to cause high mortality in P. vannamei, P. schmitti, and P. seiferus, while sparing P. stylirostris, P. monodon, P. japanicus, P. duorarum, P. chinensis, and P. aztecus (Brock 1997, Overstreet et al 1997.…”

Section: Introductionmentioning

confidence: 99%

Rapid and sensitive detection of Taura syndrome virus using nucleic acid-based amplification

Teng¹,

Chen²,

Wu³

et al. 2006

Dis. Aquat. Org.

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Requiring only simple heating devices, isothermal nucleic acid-based amplification (NASBA) is a potential detection platform to be developed for on-site diagnosis of aquaculture pathogens. In this report, an NASBA assay has been developed for the Taura syndrome virus (TSV), one of the most devastating RNA virus pathogens for several penaeid shrimp species. The NASBA amplicons were detected by agarose gel electrophoresis and confirmed by Northern-blotting and dot-blotting analysis, using a biotinylated TSV-specific primer. The sensitivity of the TSV NASBA coupled with dot-blotting detection was approximately 5-fold less sensitive than that of the commercially available RT-nested, PCR-based IQ2000 TSV Detection and Prevention System that was also confirmed to be more sensitive than the RT-PCR-based TSV detection protocol recommended by the OIE (Office International des Epizooties). The specificity of the TSV NASBA reaction was substantiated by the results that RNA of non-target viruses did not generate any signals. Furthermore, a simple colorimetric microtiter plate assay employing TSV-specific capture and detection primers was developed as a simple alternative approach for the detection of NASBA amplicons. Taken together, the combination of the isothermal NASBA and colorimetric solid phase-based assays should allow sensitive, straightforward, and speedy on-site detection of TSV. KEY WORDS: Nucleic acid-based amplification · NASBA · Diagnostics · IQ 2000 · Taura syndrome virus Resale or republication not permitted without written consent of the publisherDis Aquat Org 73: [13][14][15][16][17][18][19][20][21][22] 2006 2002, Tang & Lightner 2005). In the CP1 and CP2 regions, low genetic variation (0 to 0.24% for CP1 and 0 to 0.35 or 0 to 5.6% for CP2 nucleotide sequences) was observed among the isolates analyzed. Due to the low-fidelity nature of the RNA-dependent RNA polymerase, new isolates of TSV that are able to replicate freely in a new species, such as the one described by Chang et al. (2004) in Penaeus monodon, are expected to keep emerging.Sensitive and specific detection of the causative agents is a prerequisite for effective disease prevention and management. Methods developed for TSV detection and characterization include histological analyses, immunoassay, in situ hybridization, reverse transcription-polymerase chain reaction (RT-PCR), and real-time RT-PCR (Lightner 1995, Mari et al. 1998, Nunan et al. 1998, Poulos et al. 1999, Castagna et al. 2004, Tang et al. 2004). Among the methods described above, diagnosis of potential TSV carriers or TSV outbreaks at early stages relies mainly on sensitive and specific platforms, such as the nested and real-time RT-PCR (Romano et al. 1995, Nunan et al. 1998, Tang et al. 2004). However, because of the extensive technical training and expensive equipment involved, these systems have not been widely adopted by farm operators for sensitive and timely on-site diagnosis of TSV.With the advantage of requiring only simple heating devices, various isothermal n...

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Genetic variation and immunohistochemical differences among geographic isolates of Taura syndrome virus of penaeid shrimp

Cited by 34 publications

References 23 publications

Comparison of Taura syndrome virus (TSV) detection methods during chronic-phase infection in Penaeus vannamei

Comparison of Taura syndrome virus (TSV) detection methods during chronic-phase infection in Penaeus vannamei

Specific monoclonal antibodies raised against Taura syndrome virus (TSV) capsid protein VP3 detect TSV in single and dual infections with white spot syndrome virus (WSSV)

Rapid and sensitive detection of Taura syndrome virus using nucleic acid-based amplification

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