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Genes & Genomics

, Volume 41, Issue 1, pp 35–42 | Cite as

Multiplex PCR using YeaD and 16S rRNA gene to identify major pathogens in vibriosis of Litopenaeus vannamei

  • Yeong-Jong Han
  • Ara Jo
  • So-Won Kim
  • Hee-Eun Lee
  • Young Chul Kim
  • Hyun Do Jeong
  • Yung Hyun Choi
  • Suhkmann Kim
  • Hee-Jae Cha
  • Heui-Soo KimEmail author
Research Article
  • 164 Downloads

Abstract

The Vibrio species causing major diseases in Litopenaeus vannamei are Vibrio harveyi, Vibrio alginolyticus, and Vibrio parahaemolyticus. For multiplex PCR primers, YeaD was used to detect the three Vibrio species. Bioinformatic analysis such as MultiPLX and primer-BLAST was used to design stable and species-specific multiplex PCR primers. Multiplex PCR results showed clear band patterns with bands at 185 bp for V. alginolyticus, 396 bp for V. harveyi, 805 bp for V. arahaemolyticus, and 596 bp for common Vibrio species. The minimum concentration of DNA was measured by PCR; the value for V. alginolyticus was 0.1 ng, that of V. harveyi was 0.03 ng, and that of V. parahaemolyticus was 0.003 ng. Taken together, YeaD showed stability and specificity in identifying Vibrio species. Our multiplex PCR amplification method is an effective and inexpensive tool for identifying Vibrio species.

Keywords

Litopenaeus vannamei Multiplex polymerase chain reaction Vibrio harveyi Vibrio alginolyticus Vibrio parahaemolyticus 

Notes

Acknowledgements

This research was a part of the project titled “Omics based on fishery disease control technology development and industrialization (20150242),” funded by the Ministry of Oceans and Fisheries, Korea.

Supplementary material

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Supplementary material 1 (PPTX 1139 KB)
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Supplementary material 3 (PPTX 655 KB)

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Copyright information

© The Genetics Society of Korea and Springer Nature B.V. 2018

Authors and Affiliations

  1. 1.Department of Biological Sciences, College of Natural SciencesPusan National UniversityBusanRepublic of Korea
  2. 2.Institute of Systems BiologyPusan National UniversityBusanRepublic of Korea
  3. 3.Department of Aquatic Life MedicinePukyong National UniversityPusanRepublic of Korea
  4. 4.Department of Biochemistry, College of Korean MedicineDongeui UniversityBusanRepublic of Korea
  5. 5.Department of Chemistry, College of Natural SciencesPusan National UniversityBusanRepublic of Korea
  6. 6.Departments of Parasitology and Genetics, College of MedicineKosin UniversityBusanRepublic of Korea

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