Journal of Microbiology and Biotechnology

The Korean Society for Microbiology and Biotechnology (한국미생물·생명공학회)
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Development of Two Quantitative Real-Time PCR Diagnostic Kits for HPV Isolates from Korea

  • Jeeva, Subbiah (Department of Microbiology, Pukyong National University) ;
  • Kim, Nam-Il (Molecular Science Division, Bioneer Corporation) ;
  • Jang, In-Kwon (National Fisheries Research and Development Institute) ;
  • Choi, Tae-Jin (Department of Microbiology, Pukyong National University)
  • Received : 2012.05.14
  • Accepted : 2012.07.31
  • Published : 2012.10.28
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Abstract

Viral pathogens, alongside other pathogens, have major effects on crustacean aquaculture. Hepatopancreatic parvovirus (HPV) is an emerging virus in the shrimp industry and has been detected in shrimp farms worldwide. The HPV genome has greater diversity than other shrimp viruses owing to its wide host range and geographical distribution. Therefore, developing diagnostic tools is essential to detect even small copy numbers from the target region of native HPV isolates. We have developed two easy to use quantitative real-time PCR kits, called Green Star and Dual Star, which contain all of the necessary components for real-time PCR, including HPV primers, using the primers obtained from the sequences of HPV isolates from Korea, and analyzed their specificity, efficiency, and reproducibility. These two kits could detect from 1 to $1{\times}10^9$ copies of cloned HPV DNA. The minimum detection limits obtained from HPV-infected shrimp were $7.74{\times}10^1$ and $9.06{\times}10^1$ copies in the Green Star and Dual Star assay kits, respectively. These kits can be used for rapid, sensitive, and efficient screening for HPV isolates from Korea before the introduction of postlarval stages into culture ponds, thereby decreasing the incidence of early development of the disease.

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References

  1. Bonami, J. R. 2008. Shrimp viruses, pp. 567-576. In B. W. J. Mahy and M. H. V. van Regenmorte (eds.). Encyclopedia of Virology, 3rd Ed. Academic Press, Oxford.
  2. Bonami, J. R., J. Mari, B. T. Poulos, and D. V. Lightner. 1995. Characterization of hepatopancreatic parvo-like virus, a second unusual parvovirus pathogenic for penaeid shrimps. J. Gen. Virol. 76: 813-817. https://doi.org/10.1099/0022-1317-76-4-813
  3. Borst, A., A. Box, and A. Fluit. 2004. False-positive results and contamination in nucleic acid amplification assays: Suggestions for a prevent and destroy strategy. Eur. J. Clin. Microbiol. 23: 289-299. https://doi.org/10.1007/s10096-004-1100-1
  4. Dorak, M. T. 2006. Real-time PCR, pp. 1-24. Taylor & Francis Group, New York.
  5. Flegel, T. 2012. Historic emergence, impact and current status of shrimp pathogens in Asia. J. Inverteb. Pathol. [In Press]
  6. Flegel, T., V. Thamavit, T. Pasharawipas, and V. Alday-Sanz. 1999. Statistical correlation between severity of hepatopancreatic parvovirus infection and stunting of farmed black tiger shrimp (Penaeus monodon). Aquaculture 174: 197-206. https://doi.org/10.1016/S0044-8486(98)00507-9
  7. Gangnonngiw, W., W. Kiatpathomchai, S. Sriurairatana, K. Laisutisan, N. Chuchird, C. Limsuwan, and T. W. Flegel. 2009. Parvo-like virus in the hepatopancreas of freshwater prawns Macrobrachium rosenbergii cultivated in Thailand. Dis. Aquat. Organ. 85: 167-173.
  8. Heid, C. A., J. Stevens, K. J. Livak, and P. M. Williams. 1996. Real time quantitative PCR. Genome Res. 6: 986-994. https://doi.org/10.1101/gr.6.10.986
  9. Jang, I. K., H. S. Han, and H. J. Lim. 2006. Viral infection rate in wild broodstock of fleshy shrimp, Fenneropenaeus chinensis from Korean waters. p. 46. Proc. World Aquaculture Soc. Firenze.
  10. Jang, I. K., K. Suriakala, B. R. Kim, and X. H. Meng. 2008. Real-times PCR quantification of white spot syndrome virus (WSSV) and hepatopancreatic parvovirus (HPV) loads in shrimp and seawaters of shrimp ponds on the west coast of South Korea. J. Fish. Sci. Technol. 11: 195-204.
  11. Jang, I. K., K. Suriakala, J. Kim, X. Meng, and T. J. Choi. 2011. A TaqMan real-time PCR assay for quantifying type III hepatopancreatic parvovirus infections in wild broodstocks and hatchery-reared postlarvae of fenneropenaeus chinensis in Korea. J. Microbiol. Biotechnol. 21: 1109-1115. https://doi.org/10.4014/jmb.1107.07009
  12. Jeeva, S., S. W. Kang, Y. S. Lee, I. Jang, H. Seo, and T. J. Choi. 2012. Complete nucleotide sequence analysis of a Korean strain of hepatopancreatic parvovirus (HPV) from Fenneropenaeus chinensis. Virus Genes 44: 89-97. https://doi.org/10.1007/s11262-011-0675-8
  13. Khawsak, P., W. Deesukon, P. Chaivisuthangkura, and W. Sukhumsirichart. 2008. Multiplex RT-PCR assay for simultaneous detection of six viruses of penaeid shrimp. Mol. Cell. Probes 22: 177-183. https://doi.org/10.1016/j.mcp.2007.12.005
  14. La Fauce, K. A., J. Elliman, and L. Owens. 2007. Molecular characterisation of hepatopancreatic parvovirus (PmergDNV) from Australian Penaeus merguiensis. Virology 362: 397-403. https://doi.org/10.1016/j.virol.2006.11.033
  15. La Fauce, K. A., R. Layton, and L. Owens. 2007. TaqMan realtime PCR for detection of hepatopancreatic parvovirus from Australia. J. Virol. Methods 140: 10-16. https://doi.org/10.1016/j.jviromet.2006.10.006
  16. Lightner, D. and R. Redman. 1985. A parvo-like virus disease of penaeid shrimp. J. Invertebr. Pathol. 45: 47-53. https://doi.org/10.1016/0022-2011(85)90048-5
  17. Lightner, D., R. Redman, B. Poulos, J. Mari, J. Bonami, and M. Shariff. 1994. Distinction of HPV-type viruses in Penaeus chinensis and Macrobrachium rosenbergii using a DNA probe. Asian Fish. Sci. 7: 267-272.
  18. Lightner, D. V. 2011. Virus diseases of farmed shrimp in the Western Hemisphere (the Americas): A review. J. Invertebr. Pathol. 106: 110-130. https://doi.org/10.1016/j.jip.2010.09.012
  19. Lightner, D. V. and R. M. Redman. 1992. Penaeid virus diseases of the shrimp culture industry of the Americas, pp. 569-588. In A. W. Fast and L. J. Lester (eds.). Marine Shrimp Culture: Principles and Practice. Elsevier Science Publishers, Amsterdam
  20. Loh, P. C. 1999. Shrimp viruses, pp. 1625-1635. In G. Allan, and G. W. Robert (eds.). Encyclopedia of Virology, 2nd Ed. Elsevier, Oxford.
  21. Loh, P. C., L. M. Tapay, Y. Lu, and E. Nadala. 1997. Viral pathogens of the penaeid shrimp. Adv. Virus Res. 48: 263-312.
  22. Manjanaik, B., K. Umesha, and I. Karunasagar. 2005. Detection of hepatopancreatic parvovirus (HPV) in wild shrimp from India by nested polymerase chain reaction (PCR). Dis. Aquat. Organ. 63: 255-259.
  23. Nimitphak, T., W. Kiatpathomchai, and T. W. Flegel. 2008. Shrimp hepatopancreatic parvovirus detection by combining loop-mediated isothermal amplification with a lateral flow dipstick. J. Virol. Methods 154: 56-60. https://doi.org/10.1016/j.jviromet.2008.09.003
  24. Panichareon, B., P. Khawsak, W. Deesukon, and W. Sukhumsirichart. 2011. Multiplex real-time PCR and high-resolution melting analysis for detection of white spot syndrome virus, yellowhead virus, and Penaeus monodon densovirus in penaeid shrimp. J. Virol. Methods 178: 16-21. https://doi.org/10.1016/j.jviromet.2011.07.010
  25. Pantoja, C. and D. Lightner. 2000. A non-destructive method based on the polymerase chain reaction for detection of hepatopancreatic parvovirus (HPV) of penaeid shrimp. Dis. Aquat. Organ. 39: 177-182.
  26. Phromjai, J., V. Boonsaeng, B. Withyachumnarnkul, and T. W. Flegel. 2002. Detection of hepatopancreatic parvovirus in Thai shrimp Penaeus monodon by in situ hybridization, dot blot hybridization and PCR amplification. Dis. Aquat. Organ. 51: 227-232.
  27. Phromjai, J., W. Sukhumsirichart, C. Pantoja, D. Lightner, and T. Flegel. 2001. Different reactions obtained using the same DNA detection reagents for Thai and Korean hepatopancreatic parvovirus of penaeid shrimp. Dis. Aquat. Organ. 46: 153-158.
  28. Ririe, K. M., R. P. Rasmussen, and C. T. Wittwer. 1997. Product differentiation by analysis of DNA melting curves during the polymerase chain reaction. Anal. Biochem. 245: 154-160. https://doi.org/10.1006/abio.1996.9916
  29. Ronken, S. and A. Mol. 2007. The effect of ascorbic acid and retinoic acid on elastin and collagen gene expression in monolayers. Internal Report BMTE 07.19, University of Technology Eindhoven, Department of Biomedical Engineering.
  30. Safeena, M. P., A. Tyagi, P. Rai, I. Karunasagar, and I. Karunasagar. 2010. Complete nucleic acid sequence of Penaeus monodon densovirus (PmDNV) from India. Virus Res. 150: 1-11. https://doi.org/10.1016/j.virusres.2010.02.005
  31. Spann, K. M., R. D. Adlard, D. A. Hudson, S. B. Pyecroft, T. C. Jones, and M. O. C. Voigt. 1997. Hepatopancreatic parvolike virus (HPV) of Penaeus japonicus cultured in Australia. Dis. Aquat. Org. 31: 239-241.
  32. Srisuk, C., P. Chaivisuthangkura, W. Sukhumsirichart, P. Sridulyakul, S. Longyant, S. Rukpratanporn, and P. Sithigorngul. 2011. Improved immunodetection of Penaeus monodon densovirus with monoclonal antibodies raised against recombinant capsid protein. Aquaculture 311: 19-24. https://doi.org/10.1016/j.aquaculture.2010.08.018
  33. Sukhumsirichart, W., P. Attasart, V. Boonsaeng, and S. Panyim. 2006. Complete nucleotide sequence and genomic organization of hepatopancreatic parvovirus (HPV) of Penaeus monodon. Virology 346: 266-277. https://doi.org/10.1016/j.virol.2005.06.052
  34. Sukhumsirichart, W., W. Kiatpathomchai, C. Wongteerasupaya, B. Withyachumnarnkul, T. Flegel, V. Boonseang, and S. Panyim. 2002. Detection of hepatopancreatic parvovirus (HPV) infection in Penaeus monodon using PCR-ELISA. Mol. Cell. Probes 16: 409-413. https://doi.org/10.1006/mcpr.2002.0435
  35. Sukhumsirichart, W., C. Wongteerasupaya, V. Boonsaeng, S. Panyim, S. Sriurairatana, B. Withyachumnarnkul, and T. W. Flegel. 1999. Characterization and PCR detection of hepatopancreatic parvovirus (HPV) from Penaeus monodon in Thailand. Dis. Aquat. Organ. 38: 1-10.
  36. Tang, K. F. J. and D. V. Lightner. 2011. Duplex real-time PCR for detection and quantification of monodon baculovirus (MBV) and hepatopancreatic parvovirus (HPV) in Penaeus monodon. Dis. Aquat. Organ. 93: 191-198. https://doi.org/10.3354/dao02293
  37. Tang, K. F. J., C. R. Pantoja, and D. V. Lightner. 2008. Nucleotide sequence of a Madagascar hepato-pancreatic parvovirus (HPV) and comparison of genetic variation among geographic isolates. Dis. Aquat. Organ. 80: 105.
  38. Umesha, R. K., A. Uma, S. K. Otta, and I. Karunasagar. 2003. Detection by PCR of hepatopancreatic parvovirus (HPV) and other viruses in hatchery-reared Penaes monodon postlarvae. Dis. Aquat. Organ. 57: 141-146.
  39. Van der Velden, V., A. Hochhaus, G. Cazzaniga, T. Szczepanski, J. Gabert, and J. Van Dongen. 2003. Detection of minimal residual disease in hematologic malignancies by real-time quantitative PCR: Principles, approaches, and laboratory aspects. Leukemia 17: 1013-1034. https://doi.org/10.1038/sj.leu.2402922
  40. Yan, D., K. Tang, and D. Lightner. 2010. A real-time PCR for the detection of hepatopancreatic parvovirus (HPV) of penaeid shrimp. J. Fish. Dis. 33: 507-511. https://doi.org/10.1111/j.1365-2761.2010.01149.x
  41. Yang, B., X. L. Song, J. Huang, C. Y. Shi, and L. Liu. 2007. Evidence of existence of infectious hypodermal and hematopoietic necrosis virus in penaeid shrimp cultured in China. Vet. Microbiol. 120: 63-70. https://doi.org/10.1016/j.vetmic.2006.10.011
  42. Yu, C. I. and Y. L. Song. 2000. Outbreaks of Taura syndrome in Pacific white shrimp Penaeus vannamei cultured in Taiwan. Fish Pathol. 35: 21-24. https://doi.org/10.3147/jsfp.35.21

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