Bacterial virulence phenotypes of Escherichia coli and host susceptibility determine risk for urinary tract infections - PubMed
Absorptions of the hyperimmune serum were not carried out with cross-reactive bacterial strains. Since cross-reactivity occurs only in some bacterial strains and not in species, the absorptions are thought to be difficult. The bacterial antigens of urine specimens proved to be resistant to several freeze-thawing cycles and appear to be polysaccharides. This aspect is now under investigation . Merck and Co. , Inc. , sediada em Kenilworth, NJ, EUA (conhecida como MSD fora dos EUA e Canadá) é uma das maiores empresas de saúde mundiais trabalhando para que o mundo se sinta bem. Desde o desenvolvimento de novas terapias para o tratamento e prevenção de doenças até a assistência a pessoas carentes, estamos comprometidos em melhorar o bem-estar por todo o mundo. O Manual foi publicado pela primeira vez em 1899 para prestar um serviço comunitário. O legado deste excelente recurso continua na forma dos Manuais Merck nos EUA e Canadá e Manuais MSD no resto do mundo.
Obtenha mais informações sobre nosso compromisso para com o Conhecimento Médico Mundial. Immunologic methods are less frequently used to diagnose bacterial infections. However, bacterial antigens or antibodies can be detected in the biologic fluids of infected patients through such immunologic assays as counterimmunoelectrophoresis, coagglutination, latex agglutination, and immunoenzymatic assays (416). Hospital and public health laboratories in developing countries usually use electrophoresis or counterimmunoelectrophoresis systems for diagnostic purposes (15, 16). The reproducibility of the ELIEDA was considered to be satisfactory since 10 urine specimens from UTI patients gave similar positive results when tested twice on the same day, and 17 other urine specimens from UTI patients gave similar positive results when tested on three different days. A total of 244 urine specimens were collected and analyzed between February 1993 and May 1994 from outpatients who were clinically suspected of urinary tract infection. The specimens were collected at two locations in the city of São Paulo, Brazil, the Laboratory of Clinical Microbiology of the Department of Clinical Analysis of the University of São Paulo and the Hospital of the University of São Paulo. The outpatients ranged in age from 2 to 90; 75 of them were females and 25 were males. The specimens were examined by standard quantitative culture methods (2), and the results were compared with those obtained by ELIEDA. The urine specimens that were negative or positive from the quantitative urine culture were used as controls for the ELIEDA tests. The urine specimens used as positive controls were positive for E. coli; no micro organisms were isolated in the negative controls. 11. MacGowan AP, Marshall RJ, Cowling P, Reeves DS. Measurement of urinary lipopolysaccharide antibodies by ELISA as a screen for urinary tract infection. J Clin Pathol 1991;44:6163.
13. Myhre EB.
Rapid diagnosis of bacterial meningitis. Demonstration of bacterial antigen by counterimmunoelectrophoresis. Scand J Infect Dis 1974;6:237239. 6. Dirks-Go SIS, Zanen HC. Latex agglutination, counterimmunoelectrophoresis, and protein A co-agglutination in diagnosis of bacterial meningitis. J Clin Pathol 1978;31:11671171. 10. Kellog JA, Manzella JP, Seiple JW, Fortna SJ, Cook JW, Levisky JS. Efficacy of an enzyme-linked immunosorbent assay for detection of urinary tract immunoglobulins for diagnosis of urinary tract infections. J Clin Microbiol 1992;30:17111715.
Immunoblot assay (IBA) (19, 20) was used to characterize the polyclonal serum, using urine specimens from seven of the outpatients included in the study group. E. coli was isolated from two of them; three others presented Klebsiella pneumoniae, Proteus mirabilis, and Streptococcus sp. ; and the remaining two specimens were fully negative. For the IBA, 20 mL of sample buffer (Tris , pH 6. 8; SDS 2.
3 w/v; glycerol 10 v/v; b-mercaptoethanol 5 v/v) was added to 20 mL of centrifuged urine, and the preparation was boiled for 5 min. Electrophoresis was carried out on 12 SDS-polyacrilamide gel, the antigenic bands were transblotted to a mm nitrocellulose membrane (Millipore, Bedford, MA, United States), and the membrane was incubated with hyperimmune serum (1:2000 to E. coli). After washing, the membrane was incubated with peroxidase-conjugate antibody (1:2000) to rabbit immunoglobulin G (IgG) (Cappel-Organon Teknika Corp. , West Chester, PA, United States) and developed with diaminobenzidine (DAB) (20). 21. Leal-Bacelar GM, Kanamura HY, Hoshino-Shimizu S, Lima DMC, Abrantes-Lemos CP, Silva LC. Evaluation of the enzyme-linked-immuno-electro-diffusion-assay (ELIEDA) for the diagnosis of Schistosoma mansoni infection with low worm burden. Rev Inst Med Trop São Paulo 1995;37:123137. 2. Clarridge JE, Pezzlo MT, Vosti KL, Weissfeld AS. Laboratory diagnosis of urinary tract infections. Washington, D. C. : American Society for Microbiology; 1987. 1. Kunin CM. Detection, prevention and management of urinary tract infections, 4th ed. Philadelphia, PA: Lea & Febiger; 1987. p.
447. 19. Laemmli, UK. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 1970;227:600685. Cercetătorii britanici au dezvoltat un înveliș bactericid care se activează în lumina de joasă inte. . . 15. Requejo HIZ, Nascimento CMPC, Fahrat CK. Comparison of counterimmunoelectrophoresis, latex agglutination and bacterial culture for the diagnosis of bacterial meningitis using urine, serum and cerebrospinal fluid samples. Braz J Med Biol Res 1992;25:357367. 16.
Requejo HIZ, Alkmin MGA, Almeida RG, Casagrande ST, Cocozza AM, Lotufo JPB, et al. Dot-enzyme-linked immunosorbent assay (DOT-ELISA) for detection of pneumococcal polysaccharide antigens in pleural fluid effusion samples. Comparison with bacterial culture, counterimmunoelectrophoresis and latex agglutination. Rev Inst Med Trop São Paulo 1994;36:531537. 25. Davis JR, Stager CE, Araj GF. Clinical laboratory evaluation of a bacteriuria detection device for urine screening. Am J Clin Pathol 1984;81:4853. Pinon and Dropsy (14) combined counterimmunoelectrophoresis (CIE) with immunoenzymatic staining to create the so-called enzyme-linked immunoelectrodiffusion assay (ELIEDA). This assay was shown to be more sensitive than CIE in the serodiagnosis of parasitic and fungal infections (17). 24. Johnson CC. Definitions, classification and clinical presentation of urinary tract infections. Med Clin North Am 1991;75:241252. After a positive urinalysis, your doctor might prescribe Bactrim or Cipro, two antibiotics often used to treat UTIs caused by E. coli. If you’re not better after a few doses, the E. coli may be resistant to these drugs. 20. Towbin H, Staehelin T, Gordon J. Eletrophoretic transfer of proteins from polyacrilamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci 1979;76:43504354.
Symptoms of a UTI caused by E. coli
Bacteria are becoming increasingly resistant to antibiotics. Resistance occurs as bacteria naturally change to breakdown or avoid the antibiotics typically used to fight them. 12. McKenzie H, Young DN. Antibody to coliform antigens in urine samples from patients with symptoms of urinary tract infection. J Clin Pathol 1987;40:787792. 9. Geslin P, Legrand P, Squinazi F, Hausdorf M. Recherche dantigènes bactériens solubles dans divers produits pathologiques par contre-immuno-électrophorèse. Nouv Presse Med 1977;6:18531856. In urinary tract infections, screening tests can provide a more rapid presumptive diagnosis than the conventional bacteriological methods. The ELIEDA for E. coli antigen detection has some interesting features. For example, results are available within 4 hours of urine collection. In addition, the stained cellulose acetate strips can be stored for a long period of time after being made transparent.This allows results to be rechecked and retrospective studies to be carried out. A urinary tract infection (UTI) occurs when germs (bacteria) invade the urinary tract. The urinary tract is made up of your kidneys, bladder, ureters, and urethra. The ureters are the tubes connecting the kidneys to the bladder. The urethra is the tube carrying urine from the bladder to outside your body. 4. Bartram Jr. CE, Crowder JG, Beeler B, White A.
Diagnosis of bacterial diseases by detection of serum antigens by counterimmunoelectrophoresis, sensitivity, and specificity of detecting Pseudomonas and pneumococcal antigens. J Lab Clin Med 1974;83:591598.
Treatment for a UTI caused by E. coli
In one of these 5 urine specimens an association of E. coli with P. mirabilis was observed. A comparison of the results in detecting E. coli antigens by quantitative urine culture and by ELIEDA is given in Table 1. As standardized here, the ELIEDA provides data that reflect both bacterial identification and significant bacteriuria. The most accepted criterion for bacteriuria, ³ 105 CFU/mL, was taken as the reference in the present study, although this value may vary according to different individual conditions (1, 2, 24). UTIs are some of the most common infections doctors see. Most are caused by E. coli and are successfully treated with a round of antibiotics. If you have symptoms of a UTI, see a doctor. Most UTIs are uncomplicated and don’t cause any lasting harm to your urinary tract.
But UTIs that aren’t treated can progress to the kidneys, where
permanent damage can occur. In some cases, especially if you don’t
seem to be improving with treatment or you get recurrent
infections, a doctor may send your urine out to a lab to be
cultured. This can pinpoint exactly what bacteria is causing the
infection and what antibiotic effectively fights it.
After three more washings of 15 min each in PBS, the strip was then incubated in a mixture of 0. 5 mg/mL DAB and 1 mL/mL of 30 H2O2 in 0. 1 M Tris-HCl buffer, pH 7. 6, for 5 min, with shaking. Distilled water was added to stop the reaction. After being dried and made transparent, the strip was stored. 7.
Feigin RD, Wong M, Shackelford PG, Stechenberg BW, Dunkle LM, Kaplan S. Countercurrent immunoelectrophoresis of urine as well as of CSF and blood for diagnosis of bacterial meningitis. J Pediatr 1976;89:773775. Before this study, the ELIEDA had not been applied for antigen detection in bacterial infectious diseases. In the present study, this immunoenzymatic assay was standardized to detect E. coli-group antigens in urine specimens from outpatients with E. coli UTI, using a specific polyclonal antibody. This work also compared the ELIEDA to conventional bacteriologic methods. 8. Fossieck Jr. B, Craig R, Paterson PY. Counterimmunoelectrophoresis for rapid diagnosis of meningitis due to Diplococcus pneumoniae. J Infect Dis 1973;127:106109. O link selecionado levará você ao site de um terceiro. Não controlamos ou temos responsabilidade pelo conteúdo de sites de terceiros. Acknowledgments. This work was supported by the Coordenadoria de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) and partially by the Conselho Nacional de Pesquisa e Desenvolvimento (CNPq). We wish to thank Sonia R. C.
Buratto and Sueli Nonogaki, of the Faculdade de Ciências Farmacêuticas, University of São Paulo, SP, Brazil, for their technical assistance with the quantitative urine cultures and also the University Hospital and the Laboratory of Clinical Microbiology of the Department of Clinical Analysis of the University of São Paulo, SP, Brazil, for the urine specimens. The ELIEDA was negative for all 121 of the urine specimens for which the cultures had given negative results. The ELIEDA was also negative for the 44 urine specimens from which heterologous bacterial species were isolated by quantitative urine culture. The ELIEDA was positive for 3 urine specimens from which heterologous bacterial species were isolated by quantitative urine culture. The three bacteria that were cross-reactive with anti-E. coli serum were K. pneumoniae, Klebsiella oxytoca, and P. mirabilis. The more exposure a bacterium gets to an antibiotic, the more likely it is to alter itself to survive. Overuse and misuse of antibiotics make the problem worse. 26. Dalet F, Segovia T. Evaluation of a new agar in uricult-trio for rapid detection of Escherichia coli in urine. J Clin Microbiol 1995; 33:13951398. Figure 2. Carriage of putative urovirulence factors (PUFs) is enriched in both UAEC and non-UAEC… A group of 29 urine specimens stored at -20 °C showed no difference in their results even after two to five consecutive freeze-thawings over a period of 10 months. The hyperimmune serum did not change in agglutination titer (1:4 096) during this study period. E. coli serogroup O6 was grown in tryptic soy broth (Difco Laboratories, Detroit, MI, United States of America) overnight at 37 °C. In three inoculations containing 5 formalin (first inoculation, 0. 3 mL; second, 0. 5 mL; third, 1. 0 mL), a 3-month-old New Zealand male rabbit received whole culture medium bacteria and their extracellular products. In three subsequent inoculations of 2. 0 mL each, no formalin was used. Ewings (18) inoculation approach was utilized, and blood was collected 7 days after the last inoculation. The sera obtained were stored at -20 °C. The antibody titers were determined by an agglutination test on glass plate (18).
Five urine specimens in which E. coli was identified by the bacteriologic method but at concentrations £ 5 x 104 CFU/mL were negative with the ELIEDA. These results imply that these persons were either in the initial stage of E. coli infection or that their urine samples were contaminated. The significance of low E. coli counts could be elucidated only when new urine specimens are collected from the same outpatients. Un studiu realizat de institutul de cercetare Julius Kühn a descoperit că anumite bacterii rezistente pot ajunge . . . Urinary tract infection: detection of Escherichia coli antigens in human urine with an ELIEDA immunoenzymatic assayLas infecciones urinarias: detección de antígenos de Escherichia coli en orina humana con un ensayo inmunoenzimático por electrodifusión (ELIEDA) Your doctor may recommend doing a urine culture in which the E. coli from your sample will be tested against a variety of antibiotics to see which one is most effective in destroying it. You may even be prescribed a combination of antibiotics to fight the resistant bug. The present findings suggest that the ELIEDA is useful for laboratory diagnostic screening for UTI caused by E. coli.
However, this assay could be improved by concentrating urine, to obtain more sensitive results; preparing a mixture of hyperimmune serum specific to different bacterial species; and identifying and characterizing bacterial antigens in urinary excretions.
Source: https://www.healthline.com