ORIGINAL ARTICLE

Simultaneous detection of ompW and ctxA gene of Vibrio cholera isolated from ice used for marine product preservative by Duplex Polymerase Chain Reaction assay (dPCR)

Annisa Putri Cahyani, I Gede Putu Wirawan , I Dewa Made Sukrama

Annisa Putri Cahyani
Master Student of Biotechnology, Udayana University

I Gede Putu Wirawan
Laboratory of Genetic Resources and Biology Molecular, Udayana University. Email: [email protected]

I Dewa Made Sukrama
Departement of Clinical Microbiology, Faculty of Medicine Udayana University, Bali Indonesia
Online First: February 23, 2017 | Cite this Article
Cahyani, A., Wirawan, I., Sukrama, I. 2017. Simultaneous detection of ompW and ctxA gene of Vibrio cholera isolated from ice used for marine product preservative by Duplex Polymerase Chain Reaction assay (dPCR). Bali Medical Journal 6(1): 143-146. DOI:10.15562/bmj.v6i1.409


Background: Cholera is an infectious disease with symptom rice water stool diarrhea caused by Vibrio cholerae. Cholera transmitted by water or contaminated seafood. Ice usually used as a preservative marine products which vulnerable to contamination. Objective: This study aimed to determine the bacterial contamination of V. cholerae carrying the ompW and ctxA genes on ice preservatives marine product in Kedonganan Fish Market, Badung-Bali. Method: This study used 3 samples of ice that has not been used to preserve marine products and 20 samples of ice that has been used to preserve marine products. Samples were examined using bacterial culture methods and followed by dPCR assay to detect ompW and ctxA genes. Result: The results showed all samples of ice that has not been used preservative marine products are not contaminated by V. cholerae, while 16 of 20 (80%) samples of ice that had used contaminated by V. cholerae based on ompW gene and found no samples that carry ctxA gene. Conclusion: Ice agent (EA) was not contaminated by V. cholerae, while use ice sample marine product preservative contaminated with V. cholerae with 80% positive and  none of  isolates carry the gene ctxA.

References

Faruque, S.H.M., M.J. Albert and J.J. Mekalanos. 1998. Epidemiology, Genetics and Ecology of Toxigenic Vibrio cholerae. Microbiology and Molecular Biology Reviews. 62(4): 1301-1314.

Brooks, G.F., K.C. Carrol, J.S. Butel, S.A. Morse and T.A. Mietzner. 2013. Medical Microbiology 26th Edition. McGraw-Hill Companies Inc.

Dziejman, M., E. Balon, D. Byod, C.M. Fraser, J.F. Heidelberg and J.J. Mekalanos. 2002. Comparative Genomic Analysis of Vibrio cholerae Genes that Correlate with Cholera Endemic and Pandemic Diseases. Proceeding of the National Academy of Science. 99(2): 1556-1561.

Lesmana, M. 2004. Perkembangan Mutakhir Infeksi Kolera. J Kedokteran Trisakti. 23(3).

Novotny, L., L. Dvorska, A. Lorencova, V. Beran and I. Pavlik. 2004. Fish: a Potential Source of Bacterial Pathogens for Human Beings. Veterinarity Research Institute. Czech Republic.

Suzita, R., A.S. Abdulamir, F.A. Bakar and R.Son. 2009. A mini Review: Cholera Outbreak via Shelfish. American Journal of Infectious Disease. 5(1):40-47.

Heidelberg, J.F., J.A. Eisen, W.C. Nelson, R.A. Clayton, M.L. Gwinn, R.J. Dodson, D.H. Haft, E.K. Hickey, D.J. Peterson, L. Umayam, S.R. Gill, K.E. Nelson, T.D. Read, H.Â. Tettelin, D. Richardson, M.D. Ermolaeva, J. Vamathevan, S. Bass, H. Qin, I. Dargoi, P. Sellers, L. McDonald, T. Utterback, R.D. Fileismann, W.C. Nierman, O. White, S.L. Salzberg, H.O. Smith, R.R. Colwell, J.J. Mekalanos, J.C. Venter, and C.M. Fraser. 2000. DNA sequence of both chromosomes of the cholera pathogen Vibrio cholerae. Nature. 406: 477-484.

Alizadeh, J., R. Ranjbar, M. Kamali, N. Farhadi, A. Davari, and N. Sadeghifard. 2013. Cloning of Vibrio cholerae Outer membrane protein W in Pichia Pastoris. Iran J Microbiol. 5(3): 252-8.

Nandi, B., R.K. Nandy, G.B. Nair, T. Shimada, S. Mukhopadhyay, and A.C. Ghose. 2000. Rapid Method for Species-Specific Identification of Vibrio cholerae Using Primers Targeted to the Gene of Outer Membrane Protein OmpW. J. Clin. Microbiol. 38, No. 11: 4145-4151.

Tamrakar, A.K., M. Jain, A.K. Goel, D.V. Kamboj and L. Singh. 2009. Characterization of Vibrio cholerae from deep ground water in a cholera endemic area in Central India. Indian J Microbiol. 49(3): 271-275.

Hariyanto, D.S.T., I.G.N.K. Mahardika dan I.N. Wandia. 2015. Keragaman Spesies Ikan Tuna di Pasar Ikan Kedonganan Bali dengan Analisis Sekuen Kontrol Daerah Mitokondria DNA. Jurnal Veteriner. 16(3): 416-422.

Yasmon, A., Yusmaniar., A. Karuniawati and B. Bela. 2010. Simultaneous Detection of Legionella species and Legionella pneumophila by duplex PCR (dPCR) assay in Cooling Tower Samples from Jakarta Indonesia. Medical Journal of Indonesua. 19(4):223-227.

Thanananta N and T. Thanananta. 2008. Detection of E. coli in Water Using Duplex Polymerase Chain Reaction Technique. Thai Journal of Genetics. 1(2):109-113.

Chen, C.H, T. Shimada, N. Elhadi, S. Radu and M. Nishibuchi. 2004. Phenotyphic and Genotypic Characteristics and Epidemiological Signifiance of ctx Strain of Vibrio cholerae Isolated from Seafood in Malaysia. Appl. Envoron. Microbiol. 70(4):1964-1972.

Senderovich, Y. I. Izhaki, and M. Halpern. 2010. Fish as Reservoirs and Vectors of Vibrio cholerae PLoS ONE. 5(1): e8607

Hong, H., D.R. Patel, L.K. Tamm and B. van den Berg. 20. The Outer Membrane Protein ompW forms an eight-stranded β-barrel with a Hydrophobic channel. J. Biol. Chem. 281(11):7568-77.

Ramazanzadeh, R., S. Rouhi, P. Shakib, B. Shahbazi, F. Bidarpour and M. Karimi. 2015. Molecular Characterization of Vibrio cholerae Isolated from Clinical Samples in Kurdistan Province, Iran. Jundishsapur J Microbiol. 8(5):1-6.

Markov, E.Y., E.S. Kulikalova, L.Y. Urbanovich, V.S. Visnyakov and S.V. Balakhonov. 2015. Chitin and Product of Its Hydrolysis in Vibrio cholerae Ecology. Biochemistry (Moscow). 80(9):1109-1116.

Hill, V.R., N. Cohen, A.M. Kahler, J. L. Jones, C.A. Bopp, N. Marano, C.L. Tarr, N.M. Garrett, J. Boncy, A. Henry, G.A. Gómez, M. Wellman, M. Curtis, M.M. Freeman, M. Turnsek, R.A. Benner Jr, G. Dahourou, D. Espey, A. DePaola, J.W. Tappero, T. Handzel, and R.V. Tauxe. 2011. Toxigenic Vibrio cholerae O1 in Water and Seafood, Haiti. Emerging Infectious Diseases. 17, No. 11; 2147-2150.

Nishibori, T., G.C. de Vries, D. Rahardjo, E.B. Wasito, I. De, S. Kinoshita, Y. Hayashi, H. Hotta, M. Kawabata, T. Shirakawa, Y. Iijima and R. Osawa. 2011. Phenotypic and Genotypic Characterization of Vibrio cholerae Clinically Isolated in Surabaya Indonesia. Jpn. J. Infect. Dis. 64(1): 7-12.


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