Development of a diagnostic scar marker for Vibrio shilonii caused acute hepatopancreatic necrosis disease in whiteleg shrimp

Hoang Tan Quang, Pham Thi Diem Thi, Tran Thuy Lan, Nguyen Duc Huy, Nguyen Duy Quynh Tram, Nguyen Thi Thu Lien

Abstract


Background: In a previous report, we showed that Vibrio shilonii was found on whiteleg shrimp (Litopenaeus vannamei) with acute hepatopancreatic necrosis disease in Thua Thien Hue province, Vietnam. This study was performed to develop a diagnostic molecular marker generated by random amplified polymorphic DNA (RAPD) for V. shilonii rapid detection.

Methods: Pathogen Vibrio spp. were isolated from shrimps and fishes, and were identified by 16S rRNA sequencing. Genetic diversity of Vibrio strains was analysis by RAPD technique. Specific PCR product for V. shilonii was cloned and sequenced. SCAR marker was developed from specific PCR product.

Result: Twenty random primers were evaluated for RAPD to identify DNA polymorphisms between Vibrio species. The random primer OPN-06 generated a 468-bp DNA fragment specific for V. shilonii. This was then converted into a sequence-characterized amplified region (SCAR) marker designated N6-441.

Conclusion: Specific primers (Vshi-441F/Vshi-441R) amplified a unique DNA fragment in all V. shilonii isolates but not in the other Vibrio spp. This PCR assay showed significantly sensitive to the target DNA and reliably for the amplification the V. shilonii genome.

Keywords: AHPND; RAPD; SCAR; Vibrio shilonii; Vietnam


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References


Li J, Jiang H, Li L, Zhang X, Chen J. The Effect of Disease and Season to Hepatopancreas and Intestinal Mycobiota of Litopenaeus vannamei. Frontiers in Microbiology, (2019); 10889-889.

FAO (2020) Cultured aquatic species information programme: Penaeus vannamei (Boone, 1931). http://www.fao.org/fishery/culturedspecies/Penaeus_vannamei/en. Date accessed: 08/20/2020.

Thakur K, Patanasatienkul T, Laurin E, Vanderstichel R, Corsin F, et al. Production characteristics of intensive whiteleg shrimp (Litopenaeus vannamei) farming in four Vietnam Provinces. Aquaculture Research, (2018); 49(8): 2625-2632.

Huy ND, Ngoc LMT, Loc NH, Lan TT, Quang HT, et al. Isolation of Weissella cibaria from Pacific white shrimp (Litopenaeus vannamei) gastrointestinal tract and evaluation of its pathogenic bacterial inhibition. Indian Journal of Science & Technology, (2020); 13(10): 1200-1212.

Quang HT, Lan TT, Hai TTH, Yen PTH, Van TQK, et al. Genetic diversity and toxic genes analysis of Vibrio spp. isolated from white leg shrimp and marine fishes cultured in Tam Giang lagoon in Thua Thien Hue province, Vietnam. Indian Journal of Science & Technology, (2020); 13(13): 1412-1422.

Han JE, Tang KFJ, Piamsomboon P, Pantoja CR. Evaluation of a reliable non-invasive molecular test for the diagnosis of the causative agent of acute hepatopancreatic necrosis disease of shrimp. Aquaculture Reports, (2017); 5: 58-61.

Han JE, Tang KFJ, Tran LH, Lightner DV. Photorhabdus insect-related (Pir) toxin-like genes in a plasmid of Vibrio parahaemolyticus, the causative agent of acute hepatopancreatic necrosis disease (AHPND) of shrimp. Diseases of Aquatic Organisms, (2015); 113(1): 33-40.

González Y, Venegas D, Mendoza-Hernandez G, Camarena L, Dreyfus G. Na+- and H+-dependent motility in the coral pathogen Vibrio shilonii. FEMS Microbiology Letters, (2010); 312(2): 142-150.

Fernandez S, Katsuyama A, Kashiwabara AY, Madeira A, Durham A, Gruber A. Characterization of SCAR markers of Eimeria spp. of domestic fowl and construction of a public relational database (The Eimeria SCARdb). FEMS Microbiology Letters, (2004); 238(1): 183-188.

Kałużna M, Albuquerque P, Tavares F, Sobiczewski P, Puławska J. Development of SCAR markers for rapid and specific detection of Pseudomonas syringae pv. morsprunorum races 1 and 2, using conventional and real-time PCR. Applied microbiology and biotechnology, (2016); 100(8): 3693-3711.

Yang Y, Hu J, Chen F, Ding D, Zhou C. Development of a SCAR marker-based diagnostic method for the detection of the Citrus target spot pathogen Pseudofabraea citricarpa. BioMed Research International, (2018); article ID 7128903.

Aggarwal R, Gupta S, Banerjee S, Singh V. Development of a SCAR marker for detection of Bipolaris sorokiniana causing spot blotch of wheat. Canadian journal of microbiology, (2011); 57934-942.

Reid GA. Molecular cloning: A laboratory manual, 2nd edn: by J. Sambrook, E. F. Fritsch and T. Maniatis, Cold Spring Harbor Laboratory Press, 1989. $115.00 (3 vols; 1659 pages) ISBN 0 87969 309 6. Trends in Biotechnology, (1991); 9(1): 213-214.

Ramazanzadeh R, Rouhi S, Shakib P, Shahbazi B, Bidarpour F, et al. Molecular characterization of Vibrio cholerae Isolated from clinical samples in Kurdistan province, Iran. Jundishapur journal of microbiology, (2015); 8(5): e18119-e18119.

Sairkar PK, Sharma A, Shukla NP. SCAR marker for identification and discrimination of Commiphora wightii and C. myrrha. Molecular Biology International, (2016); 20161482796-1482796.

Rossi CC, Pereira MF, Langford PR, Bazzolli DMS. A BOX-SCAR fragment for the identification of Actinobacillus pleuropneumoniae. FEMS Microbiology Letters, (2014); 352(1): 32-37.

Anupama KP, Chakraborty A, Karunasagar I, Maiti B. Loop-mediated isothermal amplification assay as a point-of-care diagnostic tool for Vibrio parahaemolyticus: recent developments and improvements. Expert Review of Molecular Diagnostics, (2019); 19(3): 229-239.

Kim HJ, Ryu JO, Lee SY, Kim ES, Kim HY. Multiplex PCR for detection of the Vibrio genus and five pathogenic Vibrio species with primer sets designed using comparative genomics. BMC Microbiology, (2015); 15(1): 239-250.

Tsai YH, Chen PH, Yu PA, Chen CL, Kuo LT, Kuo CH. A multiplex PCR assay for detection of Vibrio vulnificus, Aeromonas hydrophila, methicillin-resistant Staphylococcus aureus, Streptococcus pyogenes, and Streptococcus agalactiae from the isolates of patients with necrotizing fasciitis. International Journal of Infectious Diseases, (2019); 81: 73-80.

Alramahy SK. Molecular diagnostics for Vibrio cholera based on recA gene isolated from human in Diwaniyah city. Journal of Pharmaceutical Sciences and Research, (2018); 10(5): 1125-1127.

Hänninen J, Takala J, Keinänen-Kiukaanniemi S. Quality of life in NIDDM patients assessed with the SF-20 questionnaire. Diabetes Research and Clinical Practice, (1998); 42(1): 17-27.

Senez B, Felicioli P, Moreau A, Goaziou M-FL. Quality of life assessment of type 2 diabetic patients in general medicin. Presse Médicale, (2004); 33(3): 161-166.

Kazemi-Galougahi MH, Ghaziani HN, Ardebili HE, Mahmoudi M. Quality of life in type 2 diabetic patients and related effective factors. Indian Journal of Medical Sciences (2012); 66(9-10): 230-237.

Shaheen F, Basit A, Riaz M, Fawwad A, Hakeem R, et al. Assessing health related quality of life in diabetic subjects by SF 36 questionnaire in a tertiary care diabetes unit of Karachi, Pakistan. . International Journal of Advanced Research, (2014); 6(6): 13-17.

Moreau A, Senez B, Felicioli P, Goaziou M-FL. Évaluation de la qualité de vie des patients diabétiques de type 2 en médecine générale. Presse Médicale, (2003); 17(608): 1-5.

Finkelstein MM. Body mass index and quality of life in a survey of primary care patients. Journal of Family Practice, (2000); 49(8): 734-737.

Eckert K. Impact of physical activity and bodyweight on health-related quality of life in people with type 2 diabetes. Diabetes, Metabolic Syndrome and Obesity, (2012); 5303-311.

Woodcock AJ, Julious SA, Kinmonth AL, Campbell MJ. Problems with the performance of the SF-36 among people with type 2 diabetes in general practice. Quality of Life Research, (2001); 10(8): 661-670.

Gulliford MC, Mahabir D. Relationship of health-related quality of life to symptom severity in diabetes mellitus: a study in Trinidad and Tobago. Journal of Clinical Epidemiology, (1999); 52(8): 773-780.

Larsson D, Lager I, Nilsson PM. Socio-economic characteristics and quality of life in diabetes mellitus–relation to metabolic control. Scandinavian Journal of Public Health, (1999); 27(2): 101-105.

Clouet F, Excler-Cavailher G, Christophe B, Masson F, Fasquel D. [Type 2 Diabetes and Short Form 36-items Health Survey]. Diabetes & Metabolism, (2001); 27(6): 711-717.

Ikeda A, Iso H, Toyoshima H, Fujino Y, Mizoue T, et al. Marital status and mortality among Japanese men and women: the Japan Collaborative Cohort Study. BMC Public Health, (2007); 773.Liu H. Till Death Do Us Part: Marital Status and U.S. Mortality Trends, 1986 – 2000. Journal of Marriage and Family, (2009); 71(5): 1158-1173.

Wee HL, Cheung YB, Li SC, Fong KY, Thumboo J. The impact of diabetes mellitus and other chronic medical conditions on health-related Quality of Life: is the whole greater than the sum of its parts? Health Qual Life Outcomes, (2005); 32.

Poljicanin T, Ajduković D, Sekerija M, Pibernik-Okanović M, Metelko Z, et al. Diabetes mellitus and hypertension have comparable adverse effects on health-related quality of life. BMC Public Health, (2010); 1012.

Lloyd A, Sawyer W, Hopkinson P. Impact of long-term complications on quality of life in patients with type 2 diabetes not using insulin. Value Health, (2001); 4(5): 392-400.


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