Supplemental Digital Content 1 Pulsed-field gel electrophoresis
Pulsed- field gel electrophoresis (PFGE) profiles of E. coli were determined. The DNA from E. coli
embedded in agarose plugs was digested with 40 U of restriction enzyme (Takara Bio Inc., Shiga, Japan) at 37°C for 16 h.
Salmonella Braenderup H9812 strain (ATCC® BAA664™) was used as the control strain (size
marker).PFGE was performed on a 1% agarose gel at 12°C (for 21 hours) using the CHEF- DRIII apparatus (Bio-Rad, Tokyo, Japan). The gel was stained with ethidium bromide(0.5μg/ml) and was observed under UV-transillumination.
Supplemental Digital Content 2 VF genes and phylogenetic grouping genes for PCR profiling Genes Virulence factors Size of the product (bp)
afa afimbrial adhesion 750
aer aerobactin 602
cnf1 cytotoxic necrotizing factor 1 498
sfa/foc S/F1C fimbriae 410
pap P fimbriae 336
fimH type 1 fimbriae adhesion 207
PAI pathogenic island marker of CFT073 930
ETTT type III secretion system 783
iroN catecholate siderophore receptor 665 iha iron-regulated gene A homolog adhesin 827 usp uropathogenic-specific protein 615
kpsMT group 2 capsule 400
traT serum resistance associates 290
fyuA ferric yersiniabactin uptake receptor 880
cvaC colicin V 680
hly alpha-hemolysin 1177
ompT outer membrane protease T 559
M 1 2 3 4 5 6 7 8 9 10 M
(Kbp)
-452.7 -398.4 -310.1 -216.9 -138.9 -104.5 -78.2 -54.7 -33.3
Supplemental Digital Content 3 Pulsed-field gel electrophoresis-profiles of Escherichia coli isolates from five infant- mother pairs
M, Molecular marker (Salmonella Braenderup); Lanes 1 and 2, infant-mother in case 2; Lanes 3 and 4, infant-mother in case 3); Lanes 5 and 6, infant-mother in case 4; Lanes 7 and 8, infant-mother in case 7; Lanes 9 and 10, infant-mother in case 10.
Supplemental Digital Content 4 Amplification products from multiplex polymerase chain reaction assays for typical strains in the two groups in 7 separate reactions using primer pools 1–7
M, Molecular marker (ΦX 174-HincII digest); A, Strain isolated from a neonate with meningitis in the Escherichia. coli-positive infant group (case 3 in Table 3); B, Strain isolated from a mother in the E. coli-negative infant group; P, positive control; N, negative control. Each lane was spliced together, removing intervening extraneous lanes from the original gel. The original data from these experiments are provided as a Supplemental Digital Content 4.
a b
c d
e
Supplemental Digital Content 5 Original data from Supplemental Digital Content 2
M, Molecular marker (ΦX 174-HincII digest); A, Strain isolated from a neonate with meningitis in the Escherichia. coli- positive infant group (case 3 in Table 3); B, Strain isolated from a mother in the E. coli-negative infant group; P1-7, positive control of primer pool 1-7; N1-7, negative control of primer pool 1-7.
Primer pool 1: afa (750 bp), aer (602 bp), cnf1 (498 bp), sfa/foc (410 bp), pap (336 bp), fimH (207 bp). pool 2: PAI (930 bp), ETTT (783 bp), iroN (665 bp), chuA (279 bp), TspE4.C2 (152 bp). pool 3: iha (827 bp), usp (615 bp), kpsMT (400 bp), traT (290 bp). pool 4: fyuA (880 bp), cvaC (680 bp), yjaA (211 bp). pool 5: hly (1177 bp). pool 6:
ompT (559 bp). pool 7: ibeA (170bp).
Supplemantal Digital Content 6 Relationship between VFs and antibiotic resistance
Antibiotic Resistance
ABPC-S (n = ABPC-R (n = ESBL (n = P