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MOLECULAR AND CELLULAR PATHOGENESIS

Molecular Epidemiologic Approaches to Urinary Tract Infection Gene Discovery in Uropathogenic Escherichia coli

Lixin Zhang, Betsy Foxman, Shannon D. Manning, Patricia Tallman, Carl F. Marrs
Lixin Zhang
Department of Epidemiology, University of Michigan School of Public Health, Ann Arbor, Michigan
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Betsy Foxman
Department of Epidemiology, University of Michigan School of Public Health, Ann Arbor, Michigan
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Shannon D. Manning
Department of Epidemiology, University of Michigan School of Public Health, Ann Arbor, Michigan
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Patricia Tallman
Department of Epidemiology, University of Michigan School of Public Health, Ann Arbor, Michigan
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Carl F. Marrs
Department of Epidemiology, University of Michigan School of Public Health, Ann Arbor, Michigan
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DOI: 10.1128/IAI.68.4.2009-2015.2000
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  • Fig. 1.
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    Fig. 1.

    (a) NotI PFGE patterns of UTI isolates. Lanes 1 to 7, first-time UTI E. coli isolates 318-11, 366-11, 373-11, 383-11, 244-11, 298-11, and 277-11 collected from patients at University of Michigan (collection 1) ordered by time (September 1993 to November 1994); lanes 8 to 10, first-time UTI E. coliisolates 1011-11, 1030-11, and 1031-11 collected from patients at University of Texas—Austin (collection 1) ordered by time (May 1992 to August 1994). All isolates are positive for fim,aer, ompT, and kpsMT (virulence profile 100111000). (b) NotI PFGE patterns of fecal isolates. Lane 1, lambda. lanes 3 to 9, fecal E. coliisolates 400-62, 331-61, 320-62, 250-62, 157-62, 153-62, and 58-62 collected from women presenting at the University of Michigan student health service gynecology clinic over a 2-month period (collection 4). All isolates are positive for fim, aer,ompT, and kpsMT (virulence profile 100111000). Lane 2, isolate 1161-11 representing the common PFGE pattern representative of first-time UTI isolates with the same virulence profile. *, Note the difference between the PFGE patterns in lane 5 (the selected fecal driver isolate) and lane 2, one of the UTI samples with a PFGE pattern typical of the cluster that includes lane 2 in panel a (the selected UTI tester isolate).

  • Fig. 2.
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    Fig. 2.

    Location and distribution of sPCR fragments on 366-11 genome. The DNA bands are the result of the NotI PFGE pattern of strain 366-11. The identifiable bands are numbered according to their sizes. The band just above band 1 is the compression zoom. N, numbers of sPCR fragments mapped to each NotI band.

Tables

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  • Table 1.

    Examples of sPCR probes hybridized to pathotype 5 isolates

    Pathotype 5 virulence signaturea and groupHybridization [no. (%)] to probe:
    P5P10P13′P24′P37P64P68P69
    1000110000000
     UTI (n = 23)14 (61)13 (57)18 (78)21 (91)1 (4)1 (4)2 (9)7 (30)
     Fecal (n = 27)18 (67)17 (63)23 (85)20 (74)2 (7)1 (4)2 (7)6 (22)
    1001110000000b
     UTI (n = 15)11 (73)11 (73)13 (87)13 (87)8 (53)13 (87)12 (80)4 (27)
     Fecal (n = 30)19 (63)16 (53)20 (67)21 (70)13 (43)24 (80)20 (67)4 (13)
    1100110000100
     UTI (n = 13)13 (100)12 (92)12 (92)13 (100)0 (0)1 (8)3 (23)1 (8)
     Fecal (n = 19)18 (95)18 (95)19 (100)15 (79)1 (5)0 (0)1 (5)3 (16)
    1101110000100
     UTI (n = 25)21 (84)9 (36)8 (32)12 (48)20 (80)19 (76)18 (72)16 (64)
     Fecal (n = 8)5 (63)1 (13)2 (25)4 (50)5 (63)5 (63)6 (75)3 (38)
    All other signatures
     UTI (n = 18)11 (61)10 (56)14 (78)15 (83)4 (22)9 (50)7 (39)6 (33)
     Fecal (n = 10)6 (60)4 (40)6 (60)5 (50)2 (20)5 (50)3 (30)3 (30)
    Total pathotype 5
     UTI (n = 94)70 (74)55 (59)65 (69)74 (79)33 (35)43 (46)42 (45)34 (36)
     Fecal (n = 94)66 (70)56 (60)70 (74)65 (69)23 (24)35 (37)32 (34)19 (20)
    Probe locationc1111110241 and 42
    • ↵a Hierarchically defined groups based on the associations among all genes. Those with the strongest associations combined with known biologic evidence for physical linkage were classified as a pathotype (28). 1 and 0, presence and absence, respectively, of the following genes (in order):fim, prf, sfa, aer,kpsMT, ompT, hly, cnfl,drb, capIII, papGad,prsGj96, and papGj96.

    • ↵b Virulence signature of tester and driver strains.

    • ↵c PFGE band of tester (366-11) DNA cleaved withNotI and hybridized.

  • Table 2.

    Hybridization of eight sPCR probes with genomic DNA from urinary and fecal E. coli isolates

    sPCR probeHybridization [no. (%)]aPb
    UTI (n = 350)Fecal (n = 349)
    P13′222 (64)149 (43)<0.0001
    P37206 (59)123 (35)<0.0001
    P69113 (32)89 (26)0.047
    P5257 (74)218 (63)0.002
    P10207 (59)129 (37)<0.0001
    P24′268 (77)192 (55)<0.0001
    P64203 (58)114 (33)<0.0001
    P68218 (62)134 (38)<0.0001
    • ↵a Urinary isolates are from women with first-time UTI; fecal isolates are from women without UTI infections (28).

    • ↵b Comparison between UTI and fecal strains using chi-square test.

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Molecular Epidemiologic Approaches to Urinary Tract Infection Gene Discovery in Uropathogenic Escherichia coli
Lixin Zhang, Betsy Foxman, Shannon D. Manning, Patricia Tallman, Carl F. Marrs
Infection and Immunity Apr 2000, 68 (4) 2009-2015; DOI: 10.1128/IAI.68.4.2009-2015.2000

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Molecular Epidemiologic Approaches to Urinary Tract Infection Gene Discovery in Uropathogenic Escherichia coli
Lixin Zhang, Betsy Foxman, Shannon D. Manning, Patricia Tallman, Carl F. Marrs
Infection and Immunity Apr 2000, 68 (4) 2009-2015; DOI: 10.1128/IAI.68.4.2009-2015.2000
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KEYWORDS

Escherichia coli
Sequence Analysis, DNA
Urinary Tract Infections

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