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The Epidemiology of Non-Enteric Escherichia Coli Infections: Prevalence of Serological Groups

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The Epidemiology of Non-Enteric Escherichia Coli Infections: Prevalence of Serological Groups THE EPIDEMIOLOGY OF NON-ENTERIC ESCHERICHIA COLI INFECTIONS: PREVALENCE OF SEROLOGICAL GROUPS Marvin Turck, Robert G. Petersdorf J Clin Invest. 1962;41(9):1760-1765. https://doi.org/10.1172/JCI104635. Research Article Find the latest version: https://jci.me/104635/pdf Journal of Clinical Investigation Vol. 41, No. 9, 1962 THE EPIDEMIOLOGY OF NON-ENTERIC ESCHERICHIA COLI INFECTIONS: PREVALENCE OF SEROLOGICAL GROUPS * By MARVIN TURCK AND ROBERT G. PETERSDORF WITH TIlE TECHNICAL ASSISTANCE OF MARY R. FOURNIER (From the Department of Medicine, University of Washington School of Medicine, and King County Hospital, Seattle, Washington) (Submitted for publication April 16, 1962; accepted May 17, 1962) Escherichia coli is an ubiquitous microorganism that the majority of non-enteric E. coli infections which is found in the gastrointestinal tract of are caused by strains of a few specific serological every individual, where it usually forms a part of groups, but did not support the idea that specific the normal gut flora. Extensive epidemiological, strains have a marked predilection for renal tis- clinical, and bacteriological observations have sue (7). This report is an extension of these documented the pathogenic significance of certain original observations and offers additional evi- serological strains of E. coli in infantile diarrhea. dence that certain serological groups of E. coli However, although they are frequently isolated in are responsible for the majority of non-enteric in- infected sites closely related to the gastrointestinal fections because of their increased prevalence in tract, such as the appendix, gall bladder, and the environment. peritoneal cavity, little is known about the sero- logical specificity of coliform bacteria in non-en- METHODS teric infections, particularly those involving the Organisms urinary tract. Ewing (1) has emphasized that The organisms included in this study were obtained complete serological typing of E. coli should pro- from the following sources: 1) Five hundred and twenty- vide accurate information concerning the incidence two strains of E. coli were isolated from patients with of specific strains associated with disease and per- non-enteric infections hospitalized at King County Hos- mit evaluation of nosocomial spread. pital (KCH) and University of Washington Hospital (UW) in Seattle, Johns Hopkins Hospital (JH) in The concept that some strains of gram-negative Baltimore, and Salt Lake General Hospital (SL) in organisms may be associated with infection more Salt Lake City. The isolates obtained from KCH were often than others is not new. Kauffman (2) ad- from consecutive patients in whom E. coli was of known vanced the hypothesis that certain coliform sero- pathogenic significance (292 strains from 276 patients). types are more prevalent in appendicitis, and he The majority of strains was of urinary origin. 2) Eighty-seven strains were cultured from urine samples also noted that strains of certain serological groups of consecutive catheterized pregnant women with low were more frequently isolated from the urine than bacterial counts (< 1,000 bacilli per ml). These were from feces. Others have reported that strains of considered to represent urethral contamination. 3) certain serological groups are more commonly Thirty-five strains were isolated from consecutive un- isolated from infected sites than from fecal speci- treated patients hospitalized with overt urinary tract in- mens (3). Vahlne (4) and Sjdstedt (5) per- fections characterized by significant bacteriuria (100,000 bacilli per ml), from whom simultaneous stool specimens formed serological typing on strains of E. coli were obtained by rectal swab. from a variety of normal and abnormal sources and found that certain types were more commonly Bacteriology associated with human infections but that these A modified method of Edwards and Ewing (8) was types were not limited to the urinary tract. More used for group differentiation of Enterobacteriaceae by biochemical tests. All strains were tested with indol, recently, Rantz (6) suggested that E. coli with methyl red, Voges-Proskauer, and Simmons' citrate certain group specific antigens are more inva- reagents. The initial 250 strains and all subsequent sive for the urinary tract than others. A pre- strains with atypical "IMVIC" patterns were further liminary report from this laboratory indicated tested with nine carbohydrates (glucose, lactose, sucrose, mannitol, salicin, inositol, adonitol, sorbitol, and dulcitol) * This work was supported in part by Training Grant for acid and gas production and with three amino acids 2E-146 (C2) of the United States Public Health Service. (lysine, arginine, and ornithine) for acid production; 1760 PREVALENCE OF SEROLOGICAL GROUPS OF ESCHERICHIA COLI 1761 they were also tested for hydrogen sulfide and urea pro- from 1: 40 to 1: 20,480 and were tested against diluted duction and for motility. As reported previously, there antigen. The antigen-antiserum mixture was incubated was no correlation between serological groups of E. coli at 500 C overnight. The highest dilution of antiserum and ability to ferment specific carbohydrates (7). showing significant agglutination was recorded. All titers 1: 1,280 were considered significant. Owing to cross- Serology reactions between various 0 groups, some strains showed Grouping sera were made available by the Communi- significant titers with more than one antiserum. Cross- cable Disease Center (CDC) Diagnostic Reagents Lab- absorbed sera were not available to determine the spe- oratory, United States Public Health Service. These cific 0 groups of these strains. are unabsorbed sera prepared by the immunization of rabbits with known 0 groups of E. coli. Complete RESULTS serological typing of E. coli cultures is dependent upon determination of the 0 group followed by characteriza- Prevalence of 0 groups in non-enteric infections. tion of K (envelope or capsular) and H (flagellar anti- The somatic antigen (O Group) of 522 strains of gens). This study is limited to determination of the 0 E. coli isolated from patients with urinary tract in- (somatic) antigen. 0 antisera for 137 standard strains of E. coli were used in this study. A modification of the fections, bacteremia, pulmonary infections, and scheme for determination of the 0 antigen described by other non-enteric purulent foci was identified Edwards and Ewing (8) was used. It may be sum- (Table I). Strains of E. coli of 54 different 0 marized as follows. groups were encountered, but four distinct sero- 1. Preparation of antigen. A trypticase soy broth cul- 4, 6, and 75-accounted for ture which had been incubated overnight at 370 C was logical groups-1, used in preparation of the E. coli antigen. The broth 44.8 per cent of all isolates. Other serological culture was heated at 1000 C for one hour in a water bath groups of particular frequency were 07, 025, 050, to destroy the thermolabile K antigen which may inhibit and the inter-O group cross-reaction of 050 with agglutination. If the bacterial suspension appeared gran- 01. A total of 401 of the 522 strains (76.8 per ular or flocculent after heating, it was defined as rough. be characterized with available All rough cultures were discarded and an additional broth cent) could typing culture was prepared from the same strain of E. coli after sera. Of the ungroupable strains, 8.8 per cent it had been serially transferred on blood agar for two suc- could not be characterized serologically because cessive days. If this culture remained rough after heat- the strains were rough, and 14.4 per cent of isolates ing, no further attempt was made to type the particular strain and it was reported as ungroupable. TABLE I 2. Agglutination with pooled specific 0 antisera. The prepared antigens (smooth colonies) of E. coli were first Prevalence of serological groups of Escherichia coli isolated man tested with pools containing combinations of the 138 from non-enteric infections of available 0 antisera according to the procedure de- scribed by Ewing, Tatum, Davis and Reavis (9, 10). 0 group No. isolated Per cent After testing the first 150 strains in our laboratory, a 1 27 5.2 separate pool containing 0 antisera 1, 4, 6, 50, and 75 4 48 9.2 was made and subsequently all strains were first tested 6 104 20.0 75 54 10.4 with this pool. If no agglutination occurred, the strain 7 15 2.9 was then tested with eight pools containing the other 133 25 13 2.5 0 antisera. Individual sera were diluted 1: 50, and after 50 12 2.3 antigen was added, each antiserum in the pool was pres- 50or 1 11 2.1 Misc. cross-reactions * 33 6.3 ent in final concentration of 1: 500. Test tubes (100 x 42 Misc. 0 groups t 84 15.9 12 mm) containing prepared antigen and pooled antisera 0 group undetermined t 75 14.4 were incubated at 50° C overnight. If there was still no Rought 46 8.8 agglutination with any of the pools, the broth culture Total groupable 401 76.8 was autoclaved at 1210 C for 2 hours in an attempt to Ungroupablet 121 23.2 destroy the A variety of K antigen. The autoclaved cul- ture was then tested with the pooled antisera, and if ag- Total 522 100.0 glutination did not occur, the strain was reported as * E. coli, 0 group undetermined (ungroupable smooth Agglutination in 2 or more sera at dilution of 1: 1,280 or strain). If there was agglutination in a pool, the antigen greater. t 0 groups 3, 5, 9, 10, 11, 12, 14, 15, 16, 18ac, 20, 21, 22, was tested with individual components contained in the 23, 26, 28ac, 29, 30, 39, 42, 43, 51, 55, 59, 60, 62, 82, 83, 85, pool in the same manner described above. 86, 91, 93, 101, 102, 110, 113, 117, 120, 121, 125ab, x2, and 3. Titrations zwith specific 0 antisera.
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