Publications by authors named "Jyh-Ming Tsai"

14 Publications

  • Page 1 of 1

sp. nov., isolated from a freshwater pond.

Int J Syst Evol Microbiol 2020 Nov 5;70(11):5899-5910. Epub 2020 Oct 5.

Laboratory of Microbiology, Department of Seafood Science, National Kaohsiung University of Science and Technology, No. 142, Hai-Chuan Rd. Nan-Tzu, Kaohsiung City 811, Taiwan, ROC.

A novel bacterial strain, designated CSW-10, isolated from a freshwater pond in Taiwan, was characterized using a polyphasic taxonomic approach. Cells were Gram-stain-negative, aerobic, non-motile, rod-shaped and formed yellow-coloured colonies. Optimal growth occurred at 30 °C, pH 7, and in the absence of NaCl. Phylogenetic analyses based on 16S rRNA gene sequences and coding sequences of 92 protein clusters indicated that strain CSW-10 formed a phylogenetic lineage in the genus . The 16S rRNA gene sequence similarity indicated that strain CSW-10 was most closely related to TNR-2 (97.6%). Strain CSW-10 showed 69.8-70.7% average nucleotide identity and 19.0-23.0% digital DNA-DNA hybridization identity with the strains of other related species. The major fatty acids of strain CSW-10 were summed feature 8 (C 7 and/or C 6) and C 6. The polar lipid profile consisted of phosphatidylethanolamine, phosphatidylglycerol, diphosphatidylglycerol, phosphatidyldimethylethanolamine, phosphatidylcholine, one uncharacterized sphingoglycolipid, five uncharacterized aminophospholipids, one uncharacterized phospholipid and one uncharacterized lipid. The predominant polyamines were homospermidine and spermidine. The major isoprenoid quinone was Q-10. Genomic DNA G+C content of strain CSW-10 was 62.0 mol%. On the basis of phenotypic and genotypic properties and phylogenetic inference, strain CSW-10 should represent a novel species of the genus , for which the name sp. nov. is proposed. The type strain is CSW-10 (=BCRC 81190 =LMG 31340).
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http://dx.doi.org/10.1099/ijsem.0.004491DOI Listing
November 2020

sp. nov., isolated from a freshwater mesocosm.

Int J Syst Evol Microbiol 2020 Oct 4;70(10):5243-5254. Epub 2020 Sep 4.

Department of Marine Biotechnology, National Kaohsiung University of Science and Technology, Kaohsiung City 811, Taiwan, ROC.

A bacterial strain, designated FSY-8, was isolated from a freshwater mesocosm in Taiwan and characterized using the polyphasic taxonomy approach. Cells of strain FSY-8 were aerobic, Gram-stain-negative, rod-shaped, non-motile and formed yellow coloured colonies on Reasoner's 2A agar. Growth occurred at 20-40 °C (optimum, 30-37 °C) and pH 5-7 (optimum, pH 6) and in the presence of 0-0.5 % NaCl (optimum, 0 %, w/v). The major fatty acids (>10 %) of strain FSY-8 were summed feature 8 (C 7 and/or C 6) and summed feature 3 (C 7 and/or C 6). The polar lipid profile consisted of a mixture of phosphatidylethanolamine, phosphatidylglycerol, phosphatidylcholine, phosphatidylmonomethylethanolamine, phosphatidyldimethylethanolamine, sphingoglycolipid, diphosphatidylglycerol, an uncharacterized aminophospholipid, an uncharacterized glycolipid and an uncharacterized lipid. The major polyamine was spermidine. The major isoprenoid quinone was Q-10. The DNA G+C content was 64.8 mol %. Phylogenetic analyses based on 16S rRNA gene sequences and coding sequences of 92 protein clusters indicated that strain FSY-8 formed a phylogenetic lineage in the genus . Strain FSY-8 showed 71.6-77.2 % average nucleotide identity and 19.9-22.8 % digital DNA-DNA hybridization identity with the strains of other species. On the basis of phenotypic and genotypic properties and phylogenetic inference, strain FSY-8 should be classified in a novel species of the genus , for which the name sp. nov. is proposed. The type strain is FSY-8 (=BCRC 81051=LMG 30053=KCTC 52812).
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http://dx.doi.org/10.1099/ijsem.0.004402DOI Listing
October 2020

sp. nov., isolated from a crocodile pond.

Int J Syst Evol Microbiol 2020 Sep;70(9):5141-5148

Laboratory of Microbiology, Department of Seafood Science, National Kaohsiung University of Science and Technology, Kaohsiung City 811, Taiwan, ROC.

Bacterial strain CCP-6, isolated from a freshwater pond in Taiwan, was characterized using a polyphasic taxonomy approach. Phylogenetic analyses based on 16S rRNA gene sequences and an up-to-date bacterial core gene set (92 protein clusters) indicated that strain CCP-6 is affiliated with species in the genus . Strain CCP-6 was most closely related to CCUG 44693 with a 98.9% 16S rRNA gene sequence similarity. Cells were Gram-stain-negative, aerobic, non-motile, rod-shaped and formed light pink-coloured colonies. Optimal growth occurred at 30 °C, pH 6 and in the absence of NaCl. The major fatty acids of strain CCP-6 were C 7, C and C cyclo 8. The polar lipid profile consisted of phosphatidylethanolamine, phosphatidylglycerol, phosphatidylcholine, phosphatidyldimethylethanolamine, phosphatidylmethylethanolamine, diphosphatidylglycerol, three unidentified aminophospholipids and an unidentified phospholipid. The predominant polyamine was spermidine. The major isoprenoid quinone was Q-10. The DNA G+C content of the genomic DNA was 69.3 mol%. Strain CCP-6 showed 85.8% average nucleotide identity and 14.5% digital DNA-DNA hybridization identity with CCUG 44693. On the basis of the genotypic, chemotaxonomic and phenotypic data, strain CCP-6 represents a novel species in the genus , for which the name sp. nov. is proposed. The type strain is CCP-6 (=BCRC 81095=LMG 30310=KCTC 62188).
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http://dx.doi.org/10.1099/ijsem.0.004396DOI Listing
September 2020

sp. nov., isolated from a freshwater lake.

Int J Syst Evol Microbiol 2020 Sep 4;70(9):4942-4950. Epub 2020 Aug 4.

Department of Marine Biotechnology, National Kaohsiung University of Science and Technology, Kaohsiung City 811, Taiwan, ROC.

A novel bacterial strain, designated TBM-1, isolated from a freshwater lake in Taiwan, was characterized using a polyphasic taxonomic approach. Phylogenetic analyses based on 16S rRNA gene sequences and coding sequences of 92 protein clusters indicated that strain TBM-1 formed a phylogenetic lineage in the genus . Analysis of 16S rRNA gene sequences showed that strain TBM-1 was most closely related to CCUG 30898 with 98.4 % sequence similarity. The average nucleotide identity, average amino acid identity and digital DNA-DNA hybridization values between strain TBM-1 and closely related strains of the genus were 74.4-77.5 %, 69.7-75.4 % and 19.8-21.8 %, respectively, supporting that strain TBM-1 represents a novel species of the genus . Cells were Gram-stain-negative, motile by means of a single polar flagellum, rod-shaped and formed blue colonies. Optimal growth occurred at 30 °C, pH 6 and 0 % NaCl. The predominant fatty acids of strain TBM-1 were summed feature 3 (C ω7 and/or C ω6), C ω7 and C. The polar lipid profile consisted of a mixture of phosphatidylethanolamine, phosphatidylglycerol, diphosphatidylglycerol, two uncharacterized aminophospholipids and two uncharacterized phospholipids. The main polyamine was putrescine. The major isoprenoid quinone was Q-8. The estimated genome size was 5.26 Mb, with an average G+C content of 70.0 mol%. On the basis of phenotypic and genotypic properties and phylogenetic inference, strain TBM-1 should be classified in a novel species of the genus , for which the name sp. nov. is proposed. The type strain is TBM-1 (=BCRC 81199 =LMG 31339).
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http://dx.doi.org/10.1099/ijsem.0.004363DOI Listing
September 2020

Identification and expression analysis of 19 CC chemokine genes in orange-spotted grouper (Epinephelus coioides).

Dev Comp Immunol 2019 08 20;97:1-10. Epub 2019 Mar 20.

Center of Excellence for the Oceans, National Taiwan Ocean University, Keelung, Taiwan, ROC; Department of Aquaculture, National Taiwan Ocean University, Keelung, Taiwan, ROC.

In this study, we describe 19 different CC chemokine genes from the orange-spotted grouper, Epinephelus coioides, identified by the analysis of the spleen transcriptome. Multiple sequence alignment of the 19 CC chemokines showed that although two genes, EcSCYA115 and EcSCYA117, shared 80% amino acid similarity (72% identity), the majority exhibited low similarity to each other. Phylogenetic analysis divided the 19 CC chemokines into six major groups. Tissue distribution analysis by RT-PCR showed that most of these chemokines were ubiquitously expressed in the 9 examined tissues, whereas some exhibited tissue-preferential expression patterns. For example, EcSCYA103 was preferentially expressed in fin and gill; EcSCYA109 in head kidney and spleen; EcSCYA114 in fin, gill, and liver; and EcSCYA119 in fin and stomach. Quantitative RT-PCR showed that after challenge with grouper iridovirus (GIV), four of the 19 CC chemokine genes, EcSYCA102, EcSYCA103, EcSYCA116, and EcSYCA118, were highly induced in the spleen. The expression of these four genes could also be upregulated by LPS and poly (I:C) challenges, suggesting that these four genes might be involved in immune response against invading pathogens.
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http://dx.doi.org/10.1016/j.dci.2019.03.006DOI Listing
August 2019

Determining the cleavage site for the mature antimicrobial peptide of Nile tilapia β-defensin using 2D electrophoresis, western blot, and mass spectrometry analysis.

Fish Shellfish Immunol 2017 Mar 9;62:41-46. Epub 2017 Jan 9.

Department of Marine Biotechnology, National Kaohsiung Marine University, 142 Hai-Chuan Road, Kaohsiung 81157, Taiwan.

Several proteomic techniques were used to determine the cleavage site of the mature antimicrobial peptide of Nile tilapia β-defensin. The computer-predicted Nile tilapia β-defensin (ASFPWSCLSLSGVCRKVCLPTELFFGPLGCGKGSLCCVSHFL) composed of 42 amino acids was chemically synthesized and prepared to produce an antibody for Western blotting. Total proteins from the skin of the Nile tilapia were separated on two-dimensional electrophoresis, and the spot of Nile tilapia β-defensin was recognized using Western blot analysis. It was then excised and extracted from the gel. The precise molecular mass of this spot was determined by LC-MS/MS spectrometry. Four major peptides were discovered, with molecular weights of 4293.2 Da, 4306.5 Da, 4678.9 Da, and 4715.0 Da. The calculated mass of the 40-amino-acid sequence (FPWSCLSLSGVCRKVCLPTELFFGPLGCGKGSLCCVSHFL) of Nile tilapia β-defensin starting from Phe27 and ending with Leu66 was 4293.18 Da, which completely matched the 4293.2 Da peptide that was obtained from the mass spectrometry analysis. This result confirmed that the cleavage site for the mature C-terminal Nile tilapia β-defensin is at residue Ser26-Phe27, not at Ala24-25 as predicted by computer analysis. This study provides a simple but reliable model to determine the cleavage site for a mature antimicrobial peptide.
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http://dx.doi.org/10.1016/j.fsi.2017.01.010DOI Listing
March 2017

Comparative proteomic analysis of Litopenaeus vannamei gills after vaccination with two WSSV structural proteins.

Fish Shellfish Immunol 2016 Feb 4;49:306-14. Epub 2016 Jan 4.

Department of Marine Biotechnology, National Kaohsiung Marine University, Kaohsiung, 81157, Taiwan, ROC. Electronic address:

White spot syndrome virus (WSSV) is one of the most devastating viral pathogens of cultured shrimp worldwide. Recently published papers show the ability of WSSV structural protein VP28 to vaccinate shrimp and raise protection against the virus. This study attempted to identify the joining proteins of the aforementioned shrimp quasi-immune response by proteomic analysis. The other envelope protein, VP36B, was used as the non-protective subunit vaccine control. Shrimp were intramuscularly injected with rVPs or PBS on day 1 and day 4 and then on day 7 their gill tissues were sampled. The two-dimensional electrophoresis (2-DE) patterns of gill proteins between vaccinated and PBS groups were compared and 20 differentially expressed proteins identified by mass spectrometry, some of which were validated in gill and hemocyte tissues using real-time quantitative RT-PCR. Many of identified proteins and their expression levels also linked with the shrimp response during WSSV infection. The list of up-regulated protein spots found exclusively in rVP28-vaccinated shrimp include calreticulin and heat shock protein 70 with chaperone properties, ubiquitin, and others. The two serine proteases, chymotrypsin and trypsin, were significantly increased in shrimp of both vaccinated groups compared to PBS controls. The information presented here should be useful for gaining insight into invertebrate immunity.
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http://dx.doi.org/10.1016/j.fsi.2015.12.044DOI Listing
February 2016

Immune responses of prophenoloxidase and cytosolic manganese superoxide dismutase in the freshwater crayfish Cherax quadricarinatus against a virus and bacterium.

Mol Immunol 2013 Nov 10;56(1-2):72-80. Epub 2013 May 10.

Department of Marine Biotechnology, National Kaohsiung Marine University, Kaohsiung 81157, Taiwan, ROC.

Prophenoloxidase (proPO) and cytosolic manganese superoxide dismutase (cytMnSOD) play crucial roles in crustacean innate immunity. In the present study, both of the above genes were cloned from hemocytes of the red claw crayfish Cherax quadricarinatus. A phylogenetic analysis of the amino acid sequences showed that C. quadricarinatus proPO and cytMnSOD were more closely related to the proPO and cytMnSOD of other crayfish than to those of penaeids, crabs, lobsters, or freshwater prawns. A tissue distribution analysis revealed that proPO was primarily expressed in hemocytes, gills, and the heart, while cytMnSOD was detected in all tissues examined. All of the crayfish artificially infected with white spot syndrome virus (WSSV) died within 4 days. According to a non-lethal dose, there was no mortality in crayfish when infected deliberately with Aeromonas hydrophila. Total hemocyte counts (THCs) had significantly decreased in crayfish at 48 and 72 h after infection with WSSV compared to the control group. In contrast, THCs of crayfish after A. hydrophila challenge had recovered by 48 and 72 h from a lower level at 24 h. There were similar responses in enzyme activities toward WSSV and A. hydrophila infection. Phenoloxidase (PO) and superoxide dismutase (SOD) activities per hemocyte significantly increased from 48 to 72 h compared to the control group. After WSSV challenge, expressions of proPO and cytMnSOD transcripts in hemocytes significantly decreased at 12h, then had respectively recovered and increased at 24 h. At 48-72 h, transcript levels were finally downregulated. No significant differences in the expression profiles of proPO and cytMnSOD were observed between the A. hydrophila-infected and control groups, besides the significant upregulation at 24h post-infection. These results implicate proPO and cytMnSOD in the immune response, and they presented similar expression patterns, although different defense mechanisms may exist for crayfish induced by WSSV and A. hydrophila.
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http://dx.doi.org/10.1016/j.molimm.2013.03.023DOI Listing
November 2013

Viral resistance and immune responses of the shrimp Litopenaeus vannamei vaccinated by two WSSV structural proteins.

Immunol Lett 2012 Nov-Dec;148(1):41-8. Epub 2012 Aug 13.

Department of Marine Biotechnology, National Kaohsiung Marine University, Kaohsiung 81157, Taiwan, ROC.

Although adaptive immunity is believed to exist only in higher vertebrates, recent studies showed the ability to vaccinate shrimp and other crayfish against white spot syndrome virus (WSSV). This study attempted to establish parameters of vaccination coordinated with subsequent viral challenge to gain insights into the mechanisms of the protective response of penaeid shrimp. Two WSSV envelope proteins, VP28 and VP36B, were used as subunit vaccines expressed in Escherichia coli followed by histidine-tag affinity chromatographic purification. Shrimp vaccinated with the recombinant WSSV proteins and challenged with diluted WSSV inocula were intramuscularly injected in order to give a precise load. Results of the viral challenge trials showed complete survival in the rVP28 group in contrast to the rVP36B and PBS groups which exhibited 100% mortality. But this effective protection was exclusively induced from a combination of a higher dosage of rVP28 and a lower viral challenge pressure. The innate immune parameters analyzed among the three groups revealed that rVP28-treated shrimp showed the highest activity level (p<0.05) of phenoloxidase and superoxide dismutase during the entire period of 7 days post-vaccination. But there were no significant differences (p>0.05) in mRNA abundances of the Down syndrome cell adhesion molecule (Dscam) among all groups. In addition, total hemocyte counts significantly decreased in shrimp treated with the recombinant viral proteins compared to the PBS group. These results indicated that the existence of structure- and dose-dependent protective responses and the elevated innate immunity in shrimp following a protein-based vaccination might be responsible for conferring resistance against WSSV.
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http://dx.doi.org/10.1016/j.imlet.2012.08.004DOI Listing
April 2013

Simultaneous detection of multiple fish pathogens using a naked-eye readable DNA microarray.

Sensors (Basel) 2012 29;12(3):2710-28. Epub 2012 Feb 29.

Aquaculture Division, Fisheries Research Institute, Ministry of Agriculture, Keelung 20246, Taiwan.

We coupled 16S rDNA PCR and DNA hybridization technology to construct a microarray for simultaneous detection and discrimination of eight fish pathogens (Aeromonas hydrophila, Edwardsiella tarda, Flavobacterium columnare, Lactococcus garvieae, Photobacterium damselae, Pseudomonas anguilliseptica, Streptococcus iniae and Vibrio anguillarum) commonly encountered in aquaculture. The array comprised short oligonucleotide probes (30 mer) complementary to the polymorphic regions of 16S rRNA genes for the target pathogens. Targets annealed to the microarray probes were reacted with streptavidin-conjugated alkaline phosphatase and nitro blue tetrazolium/5-bromo-4-chloro-3'-indolylphosphate, p-toluidine salt (NBT/BCIP), resulting in blue spots that are easily visualized by the naked eye. Testing was performed against a total of 168 bacterial strains, i.e., 26 representative collection strains, 81 isolates of target fish pathogens, and 61 ecologically or phylogenetically related strains. The results showed that each probe consistently identified its corresponding target strain with 100% specificity. The detection limit of the microarray was estimated to be in the range of 1 pg for genomic DNA and 10(3) CFU/mL for pure pathogen cultures. These high specificity and sensitivity results demonstrate the feasibility of using DNA microarrays in the diagnostic detection of fish pathogens.
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http://dx.doi.org/10.3390/s120302710DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3376613PMC
October 2012

Identification of the nucleocapsid, tegument, and envelope proteins of the shrimp white spot syndrome virus virion.

J Virol 2006 Mar;80(6):3021-9

Institute of Zoology, National Taiwan University, Taipei 106, Taiwan, Republic of China.

The protein components of the white spot syndrome virus (WSSV) virion have been well established by proteomic methods, and at least 39 structural proteins are currently known. However, several details of the virus structure and assembly remain controversial, including the role of one of the major structural proteins, VP26. In this study, Triton X-100 was used in combination with various concentrations of NaCl to separate intact WSSV virions into distinct fractions such that each fraction contained envelope and tegument proteins, tegument and nucleocapsid proteins, or nucleocapsid proteins only. From the protein profiles and Western blotting results, VP26, VP36A, VP39A, and VP95 were all identified as tegument proteins distinct from the envelope proteins (VP19, VP28, VP31, VP36B, VP38A, VP51B, VP53A) and nucleocapsid proteins (VP664, VP51C, VP60B, VP15). We also found that VP15 dissociated from the nucleocapsid at high salt concentrations, even though DNA was still present. These results were confirmed by CsCl isopycnic centrifugation followed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and liquid chromatography-nanoelectrospray ionization-tandem mass spectrometry, by a trypsin sensitivity assay, and by an immunogold assay. Finally, we propose an assembly process for the WSSV virion.
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http://dx.doi.org/10.1128/JVI.80.6.3021-3029.2006DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC1395449PMC
March 2006

The unique stacked rings in the nucleocapsid of the white spot syndrome virus virion are formed by the major structural protein VP664, the largest viral structural protein ever found.

J Virol 2005 Jan;79(1):140-9

Institute of Zoology, National Taiwan University, Taipei, Taiwan, Republic of China.

One unique feature of the shrimp white spot syndrome virus (WSSV) genome is the presence of a giant open reading frame (ORF) of 18,234 nucleotides that encodes a long polypeptide of 6,077 amino acids with a hitherto unknown function. In the present study, by applying proteomic methodology to analyze the sodium dodecyl sulfate-polyacrylamide gel electrophoresis profile of purified WSSV virions by liquid chromatography-mass spectrometry (LC-MS/MS), we found that this giant polypeptide, designated VP664, is one of the viral structural proteins. The existence of the corresponding 18-kb transcript was confirmed by sequencing analysis of reverse transcription-PCR products, which also showed that vp664 was intron-less. A time course analysis showed that this transcript was actively transcribed at the late stage, suggesting that this gene product should contribute primarily to the assembly and morphogenesis of the virion. Several polyclonal antisera against this giant protein were prepared, and one of them was successfully used for immunoelectron microscopy analysis to localize the protein in the virion. Immunoelectron microscopy with a gold-labeled secondary antibody showed that the gold particles were regularly distributed around the periphery of the nucleocapsid with a periodicity that matched the characteristic stacked ring subunits that appear as striations. From this and other evidence, we argue that this giant ORF in fact encodes the major WSSV nucleocapsid protein.
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http://dx.doi.org/10.1128/JVI.79.1.140-149.2005DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC538705PMC
January 2005

Genomic and proteomic analysis of thirty-nine structural proteins of shrimp white spot syndrome virus.

J Virol 2004 Oct;78(20):11360-70

Graduate Institute of Zoology, National Taiwan University, Taipei 106, Taiwan R.O.C.

White spot syndrome virus (WSSV) virions were purified from the hemolymph of experimentally infected crayfish Procambarus clarkii, and their proteins were separated by 8 to 18% gradient sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) to give a protein profile. The visible bands were then excised from the gel, and following trypsin digestion of the reduced and alkylated WSSV proteins in the bands, the peptide sequence of each fragment was determined by liquid chromatography-nano-electrospray ionization tandem mass spectrometry (LC-nanoESI-MS/MS) using a quadrupole/time-of-flight mass spectrometer. Comparison of the resulting peptide sequence data against the nonredundant database at the National Center for Biotechnology Information identified 33 WSSV structural genes, 20 of which are reported here for the first time. Since there were six other known WSSV structural proteins that could not be identified from the SDS-PAGE bands, there must therefore be a total of at least 39 (33 + 6) WSSV structural protein genes. Only 61.5% of the WSSV structural genes have a polyadenylation signal, and preliminary analysis by 3' rapid amplification of cDNA ends suggested that some structural protein genes produced mRNA without a poly(A) tail. Microarray analysis showed that gene expression started at 2, 6, 8, 12, 18, 24, and 36 hpi for 7, 1, 4, 12, 9, 5, and 1 of the genes, respectively. Based on similarities in their time course expression patterns, a clustering algorithm was used to group the WSSV structural genes into four clusters. Genes that putatively had common or similar roles in the viral infection cycle tended to appear in the same cluster.
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http://dx.doi.org/10.1128/JVI.78.20.11360-11370.2004DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC521807PMC
October 2004

Simultaneous detection of white spot syndrome virus (WSSV) and Taura syndrome virus (TSV) by multiplex reverse transcription-polymerase chain reaction (RT-PCR) in pacific white shrimp Penaeus vannamei.

Dis Aquat Organ 2002 Jun;50(1):9-12

King Car Biotechnology Industrial Co, Ltd, Aquaculture Research and Development Center, I-Lan, Taiwan, ROC.

An assay using a single-tube, 1-step multiplex reverse transcription-polymerase chain reaction (RT-PCR) was established for the simultaneous detection of white spot syndrome virus (WSSV) and Taura syndrome virus (TSV). Three primer sets, 9195 F/9992 R, 94 F2/R2, and ITS F/28S R, were mixed at a ratio of 3:1:1 to amplify specific fragments of the TSV, WSSV, and Penaeus vannamei genome, respectively, in the RT-PCR reaction. Shrimp samples were experimentally infected with WSSV and TSV. PCR-amplified products detected in the nucleic acid extraction of shrimp pleopods produced 4 kinds of results. With no virus infection, 1 fragment of 892 base pairs (bp) was amplified from a ribosomal RNA gene by primer set ITS F/28S R as an internal control. In samples only infected by WSSV or TSV, 2 fragments could be seen: either from WSSV (530 bp) plus the internal control or TSV (231 bp) plus the internal control, respectively. In cases of co-infection with both viruses, all 3 amplified products were detected simultaneously. This study is the first report of Penaeus vannamei specimens co-infected with WSSV and TSV being detected using a PCR method via experimental infection.
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http://dx.doi.org/10.3354/dao050009DOI Listing
June 2002
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