Author(s): Prajai Gandhi, Atulya M, Venkata Rao. J, Jesil Mathew A

Email(s): jesil.m@manipal.edu , jesilmathew@gmail.com

DOI: 10.5958/0974-360X.2020.00941.5   

Address: Prajai Gandhi1, Atulya M2, Venkata Rao. J3, Jesil Mathew A3*
1Zydus Cadila, Ahmedabad.
2CCS National Institute of Animal Health, Baghpat.
3Manipal College of Pharmaceutical Sciences, Manipal Academy of Higher Education, Manipal, Udupi, Karnataka, 576104.
*Corresponding Author

Published In:   Volume - 13,      Issue - 11,     Year - 2020


ABSTRACT:
Waterborne diseases usually result from ingestion of water contaminated by pathogenic microorganisms. Diarrhoea is a major symptom of these waterborne diseases, which leads to dehydration and in severe cases, the death of the patient. Many cases of death have been reported due to waterborne infections, especially in developing countries. Common pathogens causing waterborne diseases include Rota virus, Vibrio species, Shigella species and Escherichia coli. Several techniques have been developed for the diagnosis of pathogenic bacteria which are expensive, time-consuming and laborious processes. To overcome such problems, Polymerase Chain Reaction can be utilized as a tool for rapid identification of pathogens. The present study aims at developing a multiplex PCR for the simultaneous identification of waterborne pathogens belonging to the genera Vibrio, Shigella and Escherichia. The genes selected for study were found to be specific for chosen pathogenic strains. All the genes were amplified using the designed primers. The parameters optimised were found to be satisfactory for amplification of selected genes. The Multiplex PCR model developed could be a rapid and specific method of detection of waterborne pathogens.


Cite this article:
Prajai Gandhi, Atulya M, Venkata Rao. J, Jesil Mathew A. Rapid detection of Waterborne bacterial pathogens by multiplex PCR for the detection of bacterial pathogens in water samples. Research J. Pharm. and Tech. 2020; 13(11):5382-5386. doi: 10.5958/0974-360X.2020.00941.5


REFERENCES:
1.     Nishibuchi M, Hill WE, Zon G, Payne WL, Kaper JB. Synthetic oligodeoxyribonucleotide probes to detect Kanagawa phenomenon-positive Vibrio parahaemolyticus. J Clin Microbiol. 1986;23(6):1091–5.
2.     Nishibuchi M, Ishibashi M, Takeda Y, Kaper JB. Detection of the thermostable direct hemolysin gene and related DNA sequences in Vibrio parahaemolyticus and other vibrio species by the DNA colony hybridization test. Infect Immun. 1985;49(3):481–6.
3.     Bej AK, Patterson DP, Brasher CW, Vickery MCL, Jones DD, Kaysner CA. Detection of total and hemolysin-producing Vibrio parahaemolyticus in shellfish using multiplex PCR amplification of tl, tdh and trh. J Microbiol Methods. 1999;36(3):215–25.
4.     Tarr CL, Patel JS, Puhr ND, Sowers EG, Bopp CA, Strockbine NA. Identification of Vibrio isolates by a multiplex PCR assay and rpoB sequence determination. J Clin Microbiol. 2007;45(1):134–40.
5.     Boor KJ, Duncan ML, Price CW. Genetic and transcriptional organization of the region encoding the $β$ subunit of Bacillus subtilis RNA polymerase. J Biol Chem. 1995;270(35):20329–36.
6.     Bray J. Isolation of antigenically homogeneous strains of Bact. coli neapolitanum from summer diarrhoea of infants. J Pathol Bacteriol. 1945;57(2):239–47.
7.     Riley LW, Remis RS, Helgerson SD, McGee HB, Wells JG, Davis BR, et al. Hemorrhagic colitis associated with a rare Escherichia coli serotype. N Engl J Med. 1983;308(12):681–5.
8.     Konowalchuk J, Speirs JI, Stavric S. Vero response to a cytotoxin of Escherichia coli. Infect Immun. 1977;18(3):775–9.
9.     Levine MM, Xu J, Kaper JB, Lior H, Prado V, Tall B, et al. A DNA probe to identify enterohemorrhagic Escherichia coli of 0157: H7 and other serotypes that cause hemorrhagic colitis and hemolytic uremic syndrome. J Infect Dis. 1987;156(1):175–82.
10.     Spicer EK, Noble JA. Escherichia coli heat-labile enterotoxin. Nucleotide sequence of the A subunit gene. J Biol Chem. 1982;257(10):5716–21.
11.     Moseley SL, Hardy JW, Hug MI, Echeverria P, Falkow S. Isolation and nucleotide sequence determination of a gene encoding a heat-stable enterotoxin of Escherichia coli. Infect Immun. 1983;39(3):1167–74.
12.     Stacy-Phipps S, Mecca JJ, Weiss JB. Multiplex PCR assay and simple preparation method for stool specimens detect enterotoxigenic Escherichia coli DNA during course of infection. J Clin Microbiol. 1995;33(5):1054–9.
13.     Hartman AB, Venkatesan M, Oaks E V, Buysse JM. Sequence and molecular characterization of a multicopy invasion plasmid antigen gene, ipaH, of Shigella flexneri. J Bacteriol. 1990;172(4):1905–15.
14.     Thiem VD, Sethabutr O, von Seidlein L, Van Tung T, Chien BT, Lee H, et al. Detection of Shigella by a PCR assay targeting the ipaH gene suggests increased prevalence of shigellosis in Nha Trang, Vietnam. J Clin Microbiol. 2004;42(5):2031–5.
15.     Harley JP, Prescott LM. Laboratory exercises in microbiology. McGraw-Hill New York; 2005.
16.     Scaletsky IC, Silva ML, Trabulsi LR. Distinctive patterns of adherence of enteropathogenic Escherichia coli to HeLa cells. Infect Immun. 1984;45(2):534–6.
17.     Bendouah Z, Barbeau J, Hamad WA. -- Head and Neck Surgery Biofilm formation by Staphylococcus aureus and Pseudomonas aeruginosa is associated with an unfavorable evolution after surgery for chronic sinusitis and nasal polyposis. 2011;
18.     Mollet C, Drancourt M, Raoult D. rpoB sequence analysis as a novel basis for bacterial identification. Mol Microbiol. 1997;26(5):1005–11.
19.     Takeda Y. Thermostable direct hemolysin of Vibrio parahaemolyticus. Pharmacol Ther. 1982;19(1):123–46.
20.     Nataro JP, Kaper JB. Diarrheagenic escherichia coli. Clin Microbiol Rev. 1998;11(1):142–201.
21.     Altschul SF, Gish W, Miller W, Myers EW, Lipman DJ. Basic local alignment search tool. J Mol Biol. 1990;215(3):403–10.
22.     Krawetz SA. Bioinformatics methods and protocols. Humana Press; 2000.
23.     Maniatis T, Fritsch EF, Sambrook J. Molecular cloning: a laboratory manual Cold Spring Harbor. NY Cold Spring Harb Lab. 1982

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