Afshari, A., Rad, M., Seifi, H., Ghazvini, K. (2016). Genetic variation among Escherichia coli isolates from human and calves by using RAPD PCR. Iranian Journal of Veterinary Medicine, 10(1), 33-40. doi: 10.22059/ijvm.2016.57048
Asma Afshari; Mehrnaz Rad; Hesamedin Seifi; Kiarash Ghazvini. "Genetic variation among Escherichia coli isolates from human and calves by using RAPD PCR". Iranian Journal of Veterinary Medicine, 10, 1, 2016, 33-40. doi: 10.22059/ijvm.2016.57048
Afshari, A., Rad, M., Seifi, H., Ghazvini, K. (2016). 'Genetic variation among Escherichia coli isolates from human and calves by using RAPD PCR', Iranian Journal of Veterinary Medicine, 10(1), pp. 33-40. doi: 10.22059/ijvm.2016.57048
Afshari, A., Rad, M., Seifi, H., Ghazvini, K. Genetic variation among Escherichia coli isolates from human and calves by using RAPD PCR. Iranian Journal of Veterinary Medicine, 2016; 10(1): 33-40. doi: 10.22059/ijvm.2016.57048
Genetic variation among Escherichia coli isolates from human and calves by using RAPD PCR
1Department of Pathobiology, School of Veterinary Medicine, Ferdowsi University of Mashhad, Mashhad, Iran
2Center of Excellence in Ruminant Abortion and Neonatal Mortality and Department of Clinical Sciences, School of Veterinary Medicine, Ferdowsi Unversity of Mashhad, Mashhad, Iran
3Antimicrobial Resistance Research Center, Faculty of Medicine, Mashhad University of Medical Sciences, Mashhad, Iran
Abstract
Background: Various strains of Escherichia coli (E. coli) are known as major causes of intestinal and extraintestinal infections in humans and various animal species. Molecular methods are important for the identification of bacterial isolates and nucleotide sequence variations, as well as information on tracking bacterial agents related to the outbreaks, the frequency of the bacterial genetic structure, and the evolution of microbial populations. Objectives: The purpose of the present study was to evaluate the efficiency of the RAPD method to differentiate E. coli strains. Methods: In this study, 110 isolates of E. coli were analyzed by the RAPD PCR method using two 10bp oligonucleotides. These strains were isolated from humans with urinary tract infections and neonatal calves affected by diarrhea or septicemia. Results: Data analysis showed that 87.5% of human E. coli isolates were correctly classified in the human host group, while 94.3% of calf E. coli isolates were correctly placed in calf groups. It also demonstrated that 100% and 93.3% of isolates were accurately assigned to diarrheic and septicemic calf groups, respectively. ConclusionS: Genetic variation analysis indicated that the percentage of polymorphism among E. coli isolates from humans with urinary tract infections, diarrheic calves, and septicemic neonatal calves were 54.71%, 61.22%, and 62.5%, respectively.
تنوع ژنتیکی جدای های اشریشیا کلی از انسان و گوساله ها به روش RAPD-PCR
Authors [Persian]
اسما افشاری1; مهرناز راد1; حسام الدین سیفی2; کیارش قزوینی3
1گروه پاتوبیولوژی، دانشکده دامپزشکی دانشگاه فردوسی مشهد، مشهد، ایران
2قطب علمی مطالعات سقط جنین و مرگ و میر نوزاد دامهای نشخوارکننده و گروه علوم درمانگاهی دانشکده دامپزشکی دانشگاه فردوسی مشهد، مشهد، ایران
3مرکز تحقیقات مقاومت آنتی بیوتیکی، دانشکده پزشکی دانشگاه علوم پزشکی مشهد، مشهد، ایران
Abstract [Persian]
سویه‌های مختلف اشریشیا کلی به عنوان عوامل مهم عفونت‌های داخل و خارج روده‌ای در انسان و گونه‌های مختلف دامی شناخته می‌شوند. روش‌های مولکولی، برای شناسایی جدایه‌های باکتریایی، تنوع سکانس نوکلئوتیدی، اطلاعاتی در زمینه ردیابی عوامل باکتریایی دخیل در همه گیری‌ها، شناسایی تکراردر ساختار ژنتیکی و نیز شناسایی تکامل تدریجی باکتری‌ها، اهمیت دارند. هدف: هدف از این مطالعه ارزیابی میزان کارایی روش RAPD-PCR در تفکیک جدایه‌های اشریشیا کلی می‌باشد. روش‌کار: در این مطالعه 110 جدایه اشریشیا کلی از منابع مختلف به کمک دو الیگونوکلئوتید bp10 با سکانس‌های اختیاری به روش RAPD-PCR مورد بررسی قرار گرفتند. این جدایه‌ها از افراد مبتلا به عفونت ادراری، گوساله‌های نوزاد مبتلا به اسهال یا سپتی سمی، جدا شده بودند. نتایج: تجزیه و تحلیل اطلاعات نشان داد که 5/87٪ ازجدایه‌های اشریشیا کلی جدا شده از انسان به درستی در گروه میزبان انسان طبقه‌بندی شدند، درحالی که 3/94٪ از جدایه‌های اشریشیا کلی به درستی در گروه‌های مربوط به گوساله قرار گرفتند. همچنین نشان داده شد که 100٪ و 3/93٪ از جدایه‌ها به درستی به ترتیب درگروه‌های گوساله اسهالی وسپتی سمی قرار گرفتند. نتیجه‌گیری‌نهایی: آنالیز تنوع ژنتیکی نشان داد که درصد پلی مورفیسم در بین جدایه‌های اشریشیا کلی از افراد مبتلا به عفونت ادراری، گوساله‌های مبتلا به اسهال و گوساله‌های سپتی سمی به ترتیب 71/54٪، 22/61٪ و 5/62 ٪ بودند.
Keywords [Persian]
E. coli، تنوع ژنتیکی، RAPD-PCR
References
Al-Darahi, K.F., KhalidMahdi, L., Al-Naib, K.T., Jubreal, J. (2008) Molecular characterization of E. coli O157:H7 strains using Random Amplified Polymorphic DNA (RAPD). J Doh Uni. 11: 198-204.
Anand, N.N., Bukanov, T.U., Westblum, S. Kresovich., Berg, D.E. (2001) DNA diversity among clinical isolates of Helicobacter pylori detected by PCR based RAPD fingerprinting. Nucleic Acids Res. 20: 5137-5142.
Bando, S.Y., do Valle, G.R., Martinez, M.B., Trabulsi, L.R., Moreira-Filho, C.A. (1998) Characterization of entero invasive Escherichia coli and Shigella strains by RAPD analysis. FEMS Microbiol Lett. 165: 159-65.
Carvalho, V.M., Irino, K., Onuma, D., Pestana de Castro, A.F. (2007) Random amplification of polymorphic DNA reveals clonal relationships among enteropathogenic Escherichia coli isolated from non-human primates and humans. Braz J Med Res. 40: 237-24.
Gomes, A.R., Muniyappa, L., Krishnappa, G., Suryanarayana, V.V.S., Isioor, S., Prakash, B., and Hugar, P.G. (2005) Genotypic characterization of avian Escherichia coli by random amplification of polymorphic DNA. Int J Poult Sci. 4: 378-381.
Griffin, P.M., Tauxe, R.V. (1991) The epidemiology of infections caused by Escherichia coli O157:H7, other enterohemorrhagic E. coli, and the associated hemolytic uremic syndrome. Epidemiol Rev. 13: 60-98.
Kamerbeek, J., Schouls, L., Kolk, A., van Agterveld, M., van Soolingen, D., Kuijper, S., Bunschoten, A., Molhuizen, H., Shaw, R., Goyal, M., van Embden, J. (1997) Simultaneous detection and strain differentiation of Mycobacterium tuberculosis for diagnosis and epidemiology. J Clin Microbiol. 35: 907-914.
Leung, K.T., Mackereth, R., Tien, Y.C., Topp, E. (2004) A comparison of AFLP and ERIC-PCR analyses for discriminating Escherichia coli from cattle, pig, and human sources. FEMS Microbiol Ecol. 47: 111-119.
Lih-Ching, CH., Wey-Huey, SH., Chih, D. (2001) Characterization of Escherichia coli serotype O157 strains isolated in taiwan by PCR and multilocus enzyme analysis. J Food Drug Anal. 9: 12-19.
Maity, B., Guru, P.Y. (2007) Genetic diversity and molecular characterization of pathogenic Escherichia coli from different species of laboratory rodents. Indian J Biotechnol. 6: 210-215.
Mazurier, J., Wernars, T. (1992) Molecular epidemiological study of a mass outbreak caused by enteropathogenic Escherichia coli O157:H45. J Microbiol Immunol. 43: 381-4
Nowrouzian, F., Wold, A.E., Adlerberth, I. (2001) Computer-based analysis of RAPD (Random Amplified Polymorhic DNA fingerprints for typing of intestinal Eschericia coli. Mol Biol Today. 2: 5-10.
Ozbey, G., Kilic, A., Ertas, H.B., Muz, A. (2004) Random amplified polymorphic DNA (RAPD) analysis of Pasteurella multocida and Manheimia haemolytica strains isolated from cattle, sheep and goats. Vet Med Czech. 49: 65-69.
Owen, R.J., Hernandez, T.H. (1993) Chromosomal DNA fingerprinting- a new method ofspecies and identification applicable to microbial pathogens. J Med Microbiol. 30: 89-90.
Parabhu, V., Isloor, S., Balu, M., Suryanarayana, V.V., Rathnamma, D. (2010) Genotyping by Eric PCR of Escherichia coli isolated from bovine mastitis cases. Indian J Biotechnol. 9: 298-301.
Rodtong, S.,Tannock, G.W. (1993) Differentiation of Lactobacillus strains by ribotyping. Appl Environ Microbiol. 59: 3480-3484.
RostamZad, A., Zarkesh Esfahani, H., Enteshari, J. (2010) The investigation of molecular epidemiology of Shigella soneii isolated from clinical cases in Tehran using RAPD-PCR method. Sci Med J. 9: 279-289.
Saxena, S., Verma, J., Shikha., Mod, D.R. (2014) RAPD-PCR and 16S rDNA phylogenetic analysis of alkaline protease producing bacteria isolated from soil of India: Identification and detection of genetic variability. J Genet Engineer Biotechnol. 12: 27-35.
Shehata, A.I. (2008) Phylogenetic diversity of Staphylococcus aureus by random amplification of polymorphic DNA. Aust J Basic Appl Sci. 2: 858-863.
Tanaka, N.F., MacRae, M., Johnston, M., Mooney, J., Ogden, I.D. (1993) Optimizing enrichment conditions for the isolation of Escherichia coli O157 in soils by immunomagnetic separation. Lett Appl Microbiol. 34: 365-369.
Tenover, F., Arbeit, R., Goering, R. (1997) How to select and interpret molecular strain typing methods for epidemiological studies of bacterial infections: A review for healthcare epidemiologists. Infect Control Hosp Epidemiol. 18: 426-439.
Tseng, C., Ting, E., Johnson, D., Saluta, M., Dunst, R. (2001) RAPD fingerprinting as a potential means for differentiating human and animal E. coli. Life Sci News. 7: 10-11.
Wang G., Whittam, T.S., Berg, C.M., Berg, D.E. (1993) Rapd (arbitarary primer) PCR is more sensitive than multilocus enzyme electrophoresis for distinguishing related bacterial strains. Nucleic Acid Res. 21: 5930-5933.
Zahraei Salehi, T., Madani, S.A., Karimi, V., Arab Khazaeli, F. (2008) Molecular genetic differentiation of avian Escherichia coli by RAPD-PCR. Brac J Microbiol. 39: 494-497.
Zalewska ‘Piatek, B.M. (2011) Urinary tract infections of Escherichia coli strains of chaperone-usher system. Pol J Microbiol. 60: 279-285.
Top