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  3. Prevalence of Shiga toxin and intimin genes in Escherichia coli O157H7 strains isolated from urinary tract infection in Al-Shatar city.

Prevalence of Shiga toxin and intimin genes in Escherichia coli O157H7 strains isolated from urinary tract infection in Al-Shatar city.

Number of pages: 64 File Format: word File Code: 31999
Year: 2012 University Degree: Master's degree Category: Paramedical
Tags/Keywords: E.coliO157H7 bacteria - Escherichia coli 7H157O - Escherichia coli strains - intimin - Shiga toxin genes - Urinary tract infection - Urinary tract infections
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  • Summary of Prevalence of Shiga toxin and intimin genes in Escherichia coli O157H7 strains isolated from urinary tract infection in Al-Shatar city.

    Dissertation (or Thesis) to receive the master's degree "M.SC"

    Microbiology orientation

    Abstract:

    Abstract: E.coliO157H7 bacteria is one of the main causes of food poisoning and diseases such as: diarrhea, hemorrhagic colitis, syndrome Hemolytic uremia causes thrombocytopenic purpura and even death in humans. The purpose of this study is to evaluate the pathogenic genes 1stx, 2stx, eaeA in the urine samples of Al-Shatar city.

    Methodology: In this research, 100 urinary infection samples were collected from three hospitals of Al-Shatar city for 6 months, and after cultivation on special media using Multiplex PCR method, the presence of 1stx, 2stx, eaeA genes was evaluated.

    Results: Out of 100 UTI samples evaluated, 2 isolates with frequency (2%) had 2stx, eaeA genes. One isolate (1%) had 2stx gene and no sample with all three genes was observed.

    Conclusion: Since Escherichia coli 7H:157O is one of the basic concerns of human health. It should be given constant attention. Prevention methods can reduce the contamination of this bacteria.

    Key words: Escherichia coli 7H157O _ Shiga toxin genes _ urinary infection.

    Introduction:            

    Urinary Tract Infections are one of the most common infectious diseases in the whole life and It is one of the common causes of medical visits. These infections are after respiratory diseases in terms of frequency and are considered one of the major bacterial infections in industrialized and developing countries (11). UTI is one of the acute problems that millions of people face every year and it occurs in both men and women throughout life. Every year between 8 and 10 million visits to the doctor are due to urinary infections (14). In addition to being an annoying disease by itself, urinary tract infections sometimes cause other diseases as well. For example, some urinary tract infections can sometimes cause kidney failure, or kidney infection can spread in the blood and then spread throughout the body. Or some infections may lead to the formation of kidney stones in patients, all of which indicate the importance of identifying, investigating the prevalence and treatment of this disease (15). The solutions related to the diagnosis, treatment and follow-up of urinary infections are changing day by day. Therefore, efficient approaches with relatively less complexity should be recommended for these evaluations, and it will be very important for family doctors to be able to timely diagnose and treat urinary infections in patients. More than 90% of urinary infections are caused by bacteria, and in the rest of the cases, viruses, parasites and fungi will be involved. The most common causes of urinary tract infections are members of the Enterobacteriaceae family, especially Escherichia coli, followed by Enterobacter, Cerasia, Klebsiella and Proteus. Some types of gram-positive cocci will also play a significant role in the occurrence of this type of infection (14). Epidemiological studies have shown that urinary tract infections are not limited to certain ages, but are observed in all age groups, including infants, children, and adults (15). Escherichia coli is considered the most important bacterial cause of urinary tract infections. The ability of more than 150 strains of E. coli O157:H7 (E. coli) to colonize in the perineum and urinary tract or to migrate to the urinary tract is due to the presence of specific virulence factors in these strains. E. coli has 4 clinically important types, which are enterotoxigenic (ETEC), enteropathogenic (EPEC), enteroinvasive (EIEC) and enterohemorrhagic (EHEC). Enterohemorrhagic, which is also called Verotoxigenic Escherichia coli (VTEC) due to the secretion of a type of toxin, is one of the important causes of hemorrhagic and hemolytic syndromes, as well as subsequent renal failure, which can be problematic in the urinary tract and digestive system. Escherichia coli strain O157:H7 is the most important serotype of enterohemorrhagic strains, which has been introduced as one of the most pathogenic strains in humans.Escherichia coli O157:H7 strain is the most important serotype of enterohemorrhagic strains, which has been introduced as one of the major human pathogenic strains and causes several deaths every year through acute infections, especially by consuming contaminated food or contact with contaminated sources. As mentioned, enterohemorrhagic Escherichia coli strains secrete a toxin that is called verotoxin due to its ability to kill vero cells and Shiga-like toxin (SLT) due to its similarity to the neurotoxin secreted by Shigella dysentery type I, and the strains that produce it are also known as STEC (13). The gene producing verotoxin is located on the temperate bacteriophage genome and is introduced into Escherichia coli by phage transformation (13). Vero toxins are divided into two groups stx1 and stx2. By acting on ribosomal RNA, verotoxin prevents the synthesis of proteins, and types of Escherichia coli that produce this toxin cause dangerous syndromes such as hemorrhagic colitis (HC), hemolytic uremia syndrome (HUS), followed by kidney failure, especially at young ages (17, 18, and 19). Identification of these strains is usually not done routinely in laboratories due to their complexity, and researchers are trying to find simple methods for screening these bacteria. Therefore, in this study, it is tried to introduce and present faster methods with higher accuracy and sensitivity by using descriptive-cross-sectional studies, after performing classical methods of identification and diagnosis of Escherichia coli O157:H7 bacteria, by using molecular methods, while confirming the results obtained in the diagnosis.   

    Abstract

    Introduction: E.coil O157:H7 is one of the main factors leading to food toxicity and

    diseases such as diarrhea, bleeding coliris, uremic hemolytic syndrome, thrombocytopenic

    purpura and even death in man. The purpose of this study is to survey the prevalence of Escherichia coli O157:H7 virulence genes from Urinary tract infections in Aleshtar city.

    Materials and Methods:

    In this study 100 urine samples of urinary tract infections collected from 2 hospitals in Aleshtar city during 6 months. Then, isolated were identified with specific test.

    Then isolation of E.coli O157:H7 have been confirmed with the use of espacific antisera and with multiplex PCR method presence of E.coli O157:H7 virulence genes including: stx1, stx2, eaeA, hly has been analyzed.

    Results: Of 100 samples of urinary tract infection, 2 percent of the sample (2%) with genes 2stx, eaeA were. A sample (1%) have had 2stx gene and no samples were observed with all three genes.

    Conclusion: E.coli O157:H7 will continue to be an important public health concern as long as it

    contaminates human. Preventive measures may reduce the number of cattle that carry it.

    Keywords: Escherchia coli; Meat; Shiga-Toxigenic Escherichia coli; urinary tract infections.

  • Contents & References of Prevalence of Shiga toxin and intimin genes in Escherichia coli O157H7 strains isolated from urinary tract infection in Al-Shatar city.

    List:

    Introduction .. 1

    The main objectives of the project .. 3

    The specific objectives of the project .. 3

    The practical objectives of the project .. 3

    Hypotheses or research questions.   4

    The first chapter of text review (general).   5

    1-1 History).   6

    2-1 classification.   6

    3-1 Habitat (Habitat) .. 7

    4-1 Form of bacteria ..   7

    5-1 Cultivation of biochemistry traits.   7

    6-1 Tanks and ways of transferring bacteria.   8

    7-1 antigenic structure .. 9

    8-1 virulence factors .. 11

    1_9 types of Escherichia coli .. 12

    10_1 Escherichia coli bacteria O157:H7. 14

    Genes and virulence factors .. 18

    1_11 Natural habitat of Escherichia coli O157:H7.  19

    1_12 diagnosis .. 19

    13_epidemiology .. 20

    1-14 transmission .. 21

    1-15 signs and symptoms .. 22

    16-1 clinical manifestations .. 22

    1-17 treatment .. 24

    18-1 prevention .. 25

    Cooking and diet .. 26

    The second chapter is a review of past studies.     27

    The third chapter of materials and methods.     32

    1-3 materials and work methods .. 33

    1-1-3 materials .. 33

    1-1-1-3 cultivation environments.     33

    2-1-1-3 - other compounds.    33

    2-1-3 - Tools .. 34

    2-3- Statistical population .. 34

    3-3- Sample size .. 34

    4-3- How to collect the sample .   34

    3-5- Cultivation of samples 35

    6-3- Laboratory diagnosis.  35

    1-6-3- Warm coloring.  35

    2-6-3- oxidase test .. 35

    3-6-3- catalase test .. 35

    4-6-3- biochemical tests . 35

    1-4-6-3-Citrate use test. 36

    2-4-6-3- mobility test and indole production. 36

    3-4-6-3- Methyl Red (MR) test. 37

    4-4-6-3-Voges Proskauer (VP) test. 37

    5-4-6-3-TSI (Triple Suger Iron Agar) test. 37

    8-3-Maintenance of strains .. 37

    3-9-Preparation of bacteria for DNA extraction. 38

    1-9-3- Preparation of Luria liquid culture medium (LB, Luria broth). 38

    2-9-3- Cultivation of bacteria on LB medium. 38

    10-3- DNA extraction by BIONEER kit method. 38

    2-12-3- DNA extraction steps by BIONEER kit method. 39

    11-3- Polymerase chain reaction (PCR). 40

    Materials for PCR

    1-13-3 - PCR was done using these primers. 41. 2-13-3- Preparation of primers. 42

    3-13-3- PCR reaction components. 42

    4-13-3- How to perform PCR reaction. 43

    5-13-3- Preparation of master mix for PCR reaction. 43

    3-12- Electrophoresis. ..44

    1-12-3- Materials required for making agarose gel. 45

    2-12-3-Preparation of electrophoresis tank buffer. 45

    3-12- Preparation method of agarose gel. Photography of the gel. 47

    Chapter four, results.. 48

    1-4- Cultivation results and identification of clinical isolates. 60

    Source:

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    4. Kargar M and Homayun M. 2018; Investigating the prevalence of enterohemorrhagic Escherichia coli strains and their resistance to antibiotics among children under 5 years of age in Marodasht city. Journal of Medical Sciences of Islamic Azad University, (4)19: 273-268.

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    7. Hashemi M, Mohsenzadeh M, Bayat M, Shariatifar M, Bani Hassan M. Investigation of the prevalence of antibiotic resistance of E. coli O157:H7 isolated from minced meat in Mashhad. Proceedings of the 10th National Congress of Microbiology of Iran, Ilam, 1388: 221.

    8. Fanadi M, Salehi T, Mahzounieh M, Taheri F. 1390. Studying the virulence of verotoxin-producing strains of Escherichia coli isolated from raw milk and unpasteurized cheese. Journal of Veterinary Medicine Research, Volume 66, Number 3, 228-223.

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    10. Khorosh F, Zare Fars S, Mostafavizadeh K, Mubasharizadeh S, Izadi M, Junidi N. 2016. Comparison of the prevalence of organisms causing urinary tract infections in veterans with spinal cord injuries and patients with urinary catheters admitted to Alzahra Hospital in Isfahan. Military Medicine - No. 9 (4).

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    17_ Whittam TS, Wolfe ML, Selander RK, Dyer DW and Loos BG.1993 Clonal relationships among E.coli strains that cause hemorrhagic colitis and infantile diarrhea. Infect Immun; 61: 1619-162

    18_ Adwank Abu-Hassan N, Essawi T and Bdir M. 2002. Isolation and characterization of shiga toxigenic Escherichia coli strains from northern Palestine. J Med Microbiol; 51: 332-5.

    19_Chart H, Perry NT, Cheasty T and Wright PA. 2002. The kinetics of antibody production to antigens of Escherichia coli O157 with hemolytic uremic syndrome. J Med Microbiology; 51: 522-525.

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Prevalence of Shiga toxin and intimin genes in Escherichia coli O157H7 strains isolated from urinary tract infection in Al-Shatar city.
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