- Email: [email protected]
Characterization of Escherichia coli causing community acquired urinary tract infections in Mexico City
Characterization of Escherichia coli causing community acquired urinary tract infections in Mexico City Laura Belmont-Monroy, Rosa Mar´ıa Ribas-Aparicio, Armando Navarro´ Oca˜na, H. Angel Manjarrez-Hern´andez, Sandra Gavilanes-Parra, Gerardo Aparicio-Ozores, Patricia Isidra Cauich-S´anchez, Ulises Garza-Ramos, Jos´e Molina-L´opez PII: DOI: Reference:
S0732-8893(16)30377-7 doi: 10.1016/j.diagmicrobio.2016.11.006 DMB 14237
To appear in:
Diagnostic Microbiology and Infectious Disease
Received date: Revised date: Accepted date:
11 January 2016 4 November 2016 10 November 2016
Please cite this article as: Belmont-Monroy Laura, Ribas-Aparicio Rosa Mar´ıa, Navarro´ Oca˜ na Armando, Manjarrez-Hern´andez H. Angel, Gavilanes-Parra Sandra, AparicioOzores Gerardo, Cauich-S´anchez Patricia Isidra, Garza-Ramos Ulises, Molina-L´opez Jos´e, Characterization of Escherichia coli causing community acquired urinary tract infections in Mexico City, Diagnostic Microbiology and Infectious Disease (2016), doi: 10.1016/j.diagmicrobio.2016.11.006
This is a PDF file of an unedited manuscript that has been accepted for publication. As a service to our customers we are providing this early version of the manuscript. The manuscript will undergo copyediting, typesetting, and review of the resulting proof before it is published in its final form. Please note that during the production process errors may be discovered which could affect the content, and all legal disclaimers that apply to the journal pertain.
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Characterization of Escherichia coli causing community acquired urinary tract infections in Mexico City
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Laura Belmont-Monroya,b, Rosa María Ribas-Aparicioa, Armando NavarroOcañac, H. Ángel Manjarrez-Hernándezc,d, Sandra Gavilanes-Parrac,d, Gerardo Aparicio-Ozoresa, Patricia Isidra Cauich-Sáncheza, Ulises Garza-Ramose, José Molina-Lópezb,c,* a
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Departamento de Microbiología, Escuela Nacional de Ciencias Biológicas, Instituto Politécnico Nacional (IPN). Mexico City, Mexico. b Laboratorio de Patogenicidad Bacteriana, Unidad de Hemato-Oncología e Investigación, Hospital Infantil de México “Federico Gómez”. Mexico City, Mexico. c Departamento de Salud Pública, Facultad de Medicina, Universidad Nacional Autónoma de México (UNAM). Mexico City, Mexico. d Dirección de Investigación, Hospital General Dr. Manuel Gea González, Secretaría de Salud. Mexico City, Mexico. e Instituto Nacional de Salud Pública (INSP), Centro de Investigación Sobre Enfermedades Infecciosas (CISEI), Departamento de Diagnóstico Epidemiológico. Cuernavaca, Mor., Mexico. Running title: E. coli O25-ST131 clone from outpatients, UTI
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*Corresponding author: Departamento de Salud Pública, Facultad de Medicina, UNAM. México D.F. 04510, México. Phone: (52-55) 52 28 99 17. E-mail: [email protected]
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Abstract
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The O25-ST131 clone was identified within 169 uropathogenic Escherichia coli (UPEC) strains. The 44.8% of the 29 O25-ST131 clones detected were positive to least to one
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extended-spectrum β-lactamase gene. The phylogroup D was mainly found. The O25-
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ST131 clone appeared to be associated with community-acquired UTI in Mexico City.
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Keywords: Urinary infection; Escherichia coli; Multiplex PCR; Serogroups; ST131 clone
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Escherichia coli clone O25-ST131 has emerged worldwide as important cause of community-onset urinary tract infection (UTI) (Nicholas-Chanoine et al.,
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T
2008; Coque et al., 2008). This clone has been often associated with extendedspectrum -lactamases (ESBL) enzymes and is frequently Multidrug-Resistant
SC
(MDR) (Johnson et al., 2002; Rogers et al. 2011). There is no comprehensive surveillance of E. coli causing community-acquired UTI in Mexico City; therefore, it
NU
is very difficult to estimate the true incidence of UTI and to monitor the spread of
MA
this organism. The aim of this study was to determine the O-serogroup of uropathogenic E. coli (UPEC) strains, the prevalence of the clone O25-ST131 in E.
ED
coli isolates from outpatients diagnosed with an UTI, and their association with
PT
phylogroups and the presence of ESBL genes. One hundred sixty nine E. coli isolates, one isolate per patient, were
CE
collected from community-acquired UTI in Mexico. These isolates that met the
AC
inclusion criteria were selected from 800 outpatients and collected between February 2010 and October 2012, at Mexico General Hospital. The outpatient’s aged was between 18 and 70 years. For this study was consider a UTI acquired in the community that was present in apparently healthy patients who only attended the consultation by the onset of symptoms. Excluding criteria were: patients with complicated-UTI; if they were pregnant; if they had diabetes, if they were immunocompromised; or if other suspected sources of infection were present. Identification of isolates was carried out using standard microbiological techniques (Carridge et al., 1988; Devine et al., 1989). An initial serotyping as agglutination
3
ACCEPTED MANUSCRIPT 4
assay (Ørskov and Ørskov, 1984) was carried out, using 96-well microtiter plates and rabbit serum, obtained against 185 somatic antigens. Isolates that failed to
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T
agglutinate with any of these antisera were defined as Non-Typeable (NT), whereas those that did agglutinate with saline solution were termed Auto-
SC
Agglutinable (AA).
NU
Fourteen pairs of primers were used by determination of UPEC serogroups by multiplex PCR, among AA and NT strains detected by conventional serotyping.
MA
Multiplex PCR was performed according to Li et al. (2010). The phylogroups were determined by PCR assay, using a combination of three DNA markers (chuA, yjaA,
ED
and TspE4C2.1) (Clermont et al., 2000). All O25 serogroup isolates were tested to
PT
determine those of the O25-ST131 clone by multiplex PCR assay (allele-specific PCR for the pabB gene) according to Clermont et al. (2009). Finally, both Narrow-
CE
Spectrum -Lactamases (NSBL) and ESBL: TEM-1, CMY-, OXA-1, SHV- and
al. (2013).
AC
CTX-M-15 type were screened by PCR using specific primers according to Kiiru et
Of the 169 E. coli isolates, 142 (84.0%) were typeable, 12 (7.1%) were NT, and 15 (8.9%) were AA strains. Of the typeable strains, 80/142 (47.3%) belonged to UPEC serogroups and 62/142 (36.7%), to non-UPEC serogroups. The O25 serogroup was the most common with 29 (17.1%) of the isolates, followed by O1 (10%), O8 (9%), O6 (4%), and O75 (3%). The NT isolates were identified as O15, O18, O21, and O25 serogroups and AA isolates were identified as O25, O21, O18,
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and O8 serogroups. Among the O25 serogroup all corresponded to O25-ST131
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clone (28 identified by conventional serotyping and one by multiplex PCR).
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Otherwise, 64 (37.9%) of the isolates belonged to phylogroup D, 47 (27.8%) belong to group A, 30 (17.8%) to group B1, and 28 (16.5%) to group B2. We found
SC
statistically significant differences between phylogroup B2 and UPEC vs. non-
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UPEC serogroups (p value, 0.03), (Table 1). On the other hand, the majority of NT and AA isolates belonged to the A and B2 phylogroups. Twelve (41.4%) of the
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O25-ST131 clone identified corresponded to the D phylogenetic group, ten (34.4%)
ED
to B2, four (13.8%) to A, and three (10.4%) to the B1 phylogroup. Thirteen (44.8%) of the 29 O25-ST131 clones detected were positive to
PT
least to one NSBL- or ESBL-type, showed various combinations of -lactamase
CE
genes (Table 2).
The ST131 E. coli isolates are considered to be truly pathogenic, due to the
AC
spectrum of infections that they cause in both community and hospital settings and to the large number of virulence-associated genes that they contain (NicolasChanoine et al., 2014). We found that these strains cause community-acquired UTI in patients in Mexico City; including some E. coli isolates belonging to the O25ST131 clone. In this study, 29 isolates with O25 serogroup were positives to O25ST131. It has been reported that the majority of O25-ST131 isolates are MDR and that they commonly produce CTX-M-15 ESBL enzyme (Nicholas-Chanoine et al., 2008), causing UTI and significant morbidity and mortality; this clone constitutes a
5
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major public health concern (Nicholas-Chanoine et al., 2008; Clermont et al., 2009;
T
Peirano and Pitout, 2010).
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In this study, phylogenetic groups A and D were common (27.8 and 37.9%, respectively) among our isolates. Lopez-Banda et al., 2013 found a high
SC
percentage of strains belonging to the phylogenetic group A (30.6%) and B2
NU
(55.6%) and less frequently B1 (9.3%) and D (4.6%). This was not expected because extra-intestinal virulent strains belong mainly to groups D and B2; we
MA
obtained 17.8% of group-B1 strains and 16.5% of group-B2 strains. Due to that a high percentage of strains of this study belong to group A, it is necessary to
ED
continue to monitor the phylogroups considered as commensal, as in the case of
as
commensal
PT
group A, because these are capable of causing UTI in the population. Thus, its role should
be
reconsidered.
This
result
suggests
that
the
AC
the urinary tract.
CE
gastrointestinal tract is the main reservoir of strains that may be able to colonize
Recently, the E. coli ST131 lineage and phylogenetic group B2 were shown as the major clones among CTX-M-15-producing E. coli isolates obtained from several clinical sources, mainly urine and blood in Australia, Canada, India, Spain, the U.K., and New Zealand. (Petty et al., 2014). In Mexico, in strains from community acquired UTI, it was found that 13/17 isolates with serogroup O25 belonged to the O25-ST131 clone (Molina-López et al., 2011), while Reyna-Flores et al. (2013) found that 25% of 4,735 UPEC isolates belonged to the O25b-ST131
6
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clone, compared to the 17% identified in this study. These reports document the spread and success of the ST131 E. coli lineage. In our study, a low percentage of
RI P
T
ESBL was obtained as the strains of ST131 cloned, and only two strains were positive for the blaCTX-M-15 gene. The O25-ST131 strains found were not associated
SC
with the presence of ESBL; therefore, we suggest that the resistance of these strains is not mediated by these enzymes. Likewise, we demonstrated the
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prevalence of serogroup O25 strains, which aids in detecting the spread of specific
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clones, such as O25-ST131, among outpatients with UTI in Mexico City.
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Acknowledgements
This study was supported by CONACyT, through its support to project No. 166004,
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Pública, UNAM.
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and PAPIIT number IN228916, Facultad de Medicina, Departamento de Salud
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References
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Carridge JE, Johnson JR, Pezzlo MT (1988) Laboratory diagnosis of urinary tract
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infections. In: Weissfeld AS, editor. Cumitech 2B. Washington: ASM Press. p. 2-19
SC
Clermont O, Bonacorsi S, Bingen E (2000) Rapid and simple determination of the Escherichia coli phylogenetic group. Appl Environ Microbiol 66: 4555-4558.
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Clermont O, Dhanji H, Upton M, Gibreel T, Fox A, Boyd D, Mulvey MR, Nordmann
MA
P, Ruppé E, Sarthou JL, Frank T, Vimont S, Arlet G, Branger C, Woodford N, Denamur E (2009) Rapid detection of the O25b-ST131 clone of Escherichia coli
ED
encompassing the CTX-M-15-producing strains. J Antimicrob Chemother.64: 274-
PT
277.
Coque TM, Novais A, Carattoli A, Poirel L, Pitout J, Peixe L, Baquero F, Cantón R,
CE
Nordmann P (2008) Dissemination of clonally related Escherichia coli strains
195-200.
AC
expressing extended-spectrum beta-lactamase CTX-M-15. Emerg Infect Dis 14:
Devine DA, Robinson L, Roberts AP (1989) Occurrence of KI, K5 and O antigens in Escherichia coli isolates from patients with urinary tract infections or bacteraemia J Med Microbiol 30: 295-299. Johnson JR, Kuskowski MA, O'Bryan TT, Maslow JN (2002) Epidemiological correlates of virulence genotype and phylogenetic background among Escherichia
8
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coli blood isolates from adults with diverse-source bacteremia. J Infect Dis 185:
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1439-1447.
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Kiiru J, Butaye P, Goddeeris BM and Kariuki S (2013) Analysis for prevalence and physical linkages amongst integrons, ISEcp1, ISCR1, Tn21 and Tn7 encountered
SC
in Escherichia coli strains from hospitalized and non-hospitalized patients in Kenya
NU
during a 19-year period (1992–2011) BMC Microbiology 13:109-123 Li D, Liu B, Chen M, Guo D, Guo X, Liu F, Feng L, Wang L (2010) A multiplex PCR
MA
method to detect Escherichia coli serogroups associated with urinary tract
López-Banda
DA,
ED
infections. J Microbiol Methods 82:71-77. Carrillo-Casas AH,
PT
G,Manjarrez-Hernández
EM,
Leyva-Leyva
Arroyo-Escalente
S,
et
M,
Orozco-Hoyuela
al.
Identification
of
CE
virulencefactors genes in Escherichia coli isolates from women with urinary tract
AC
infectionin Mexico. Biomed Res Int. 2014; 40:1565–6. Molina-López J, Aparicio-Ozores G, Ribas-Aparicio RM, Gavilanes-Parra S, Chávez-Berrocal ME, Hernández-Castro R, Manjarrez-Hernández HA (2011) Drug resistance, serotypes, and phylogenetic groups among uropathogenic Escherichia coli including O25-ST131 in Mexico City. J Infect Dev Ctries 5:840-849. Nicholas-Chanoine MH, Blanco J, Leflon-Guibout V, Demarty R, Alonso MP, Caniça MM, Park YJ, Lavigne JP, Pitout J, Johnson JR (2008) Intercontinental emergence of Escherichia coli clone O25:H4-ST131 producing CTX-M-15. J Antimicrob Chemother 61: 273-281.
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Nicolas-Chanoine, M.-H., Bertrand, X., & Madec, J.-Y. (2014). Escherichia coli ST131, an Intriguing Clonal Group. Clinical Microbiology Reviews, 27(3), 543–574.
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T
http://doi.org/10.1128/CMR.00125-13
Ørskov F and Ørskov I. (1984). Serotyping of Escherichia coli. In Methods in
SC
Microbiology, vol. 14, pp. 43–112. Edited by T. Bergan. London: Academic Press.
NU
Peirano G, Pitout JD (2010) Molecular epidemiology of Escherichia coli producing
Antimicrob Agents 35: 316-321.
MA
CTX-M beta-lactamases: the worldwide emergence of clone ST131 O25:H4. Int J
ED
Petty NK, Zakour NLB, Stanton-Cooka,M, Skippington E, Totsika M, Forde BM, Phan MD, Moriel DG, Peters KM, Davies M, Rogers BA, Dougan G, Rodriguez-
PT
Baño J, Pascual A, Pitouth JDD, Upton M, Paterson DL, Walsh TR, Schembria
CE
MA, and Beatson SA. (2014) Global dissemination of a multidrug resistant
AC
Escherichia coli clone. PNAS 111:5694-5699. Reyna-Flores F, Barrios H, Garza-Ramos U, Sanchez-Perez A, Rojas-Moreno T, Uribe-Salas
FJ,
Facundo-Sierra
R,
Silva-Sanchez
J.
(2013)
Molecular
epidemiology of Escherichia coli O25b-ST131 isolates causing communityaquiered UTIs in Mexico. Diag Microbiol Infect Dis 76:396-398.
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Rogers, B. A., Sidjabat, H. E., & Paterson, D. L. (2011). Escherichia coli O25b-
AC
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Chemother 66: 1–14. http://doi.org/10.1093/jac/dkq415
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ST131: A pandemic, multiresistant, community-associated strain. J Antimicrob
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Table 1. Phylogenetic distribution of E. coli isolates by serogroups determined by conventional serotyping and polymerase chain reaction (PCR) P
UPEC serogroups
Non-UPEC
group
(n = 80)
serogroups (n = 62)
value
A
26
21
0.11
B1
16
B2
23
D
36
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SC
0.79
5
0.03
28
0.98
NU
14
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T
Phylogenetic
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UPEC = Uropathogenic E. coli
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Table 2. NSBL- and ESBL-genes combination present in E. coli O25-ST131
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isolates from outpatients with UTI.
NSBL- and ESBL genes
strains
combination
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Frequency in O25-ST131
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N (%) TEM-1a
2 (6.7)
TEM-1a/OXA-1a
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7 (24.1)
OXA-1a
1 (3.5)
CTX-M-15b
ED
1 (3.5)
CTX-M-15b/TEM-1a
1 (3.5)
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16 (55.2)
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1 (3.5)
CMY Negative to -lactamase genes
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NSBL: Narrow-Spectrum -Lactamase
a
ESBL: Extended-Spectrum -Lactamase
b
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Highlights
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The O25-ST131 strains found were not associated with the presence of ESBL.
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The E. coli O25-ST131 isolates negative for -lactamase genes were higher in
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comparison with the presence of NSBL or ESBL genes
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The prevalence of serogroup O25 strains, aids in detecting the spread the O25-
AC
CE
PT
ED
ST131 clone among outpatients with UTI in Mexico City.
14
S0732-8893(16)30377-7 doi: 10.1016/j.diagmicrobio.2016.11.006 DMB 14237
To appear in:
Diagnostic Microbiology and Infectious Disease
Received date: Revised date: Accepted date:
11 January 2016 4 November 2016 10 November 2016
Please cite this article as: Belmont-Monroy Laura, Ribas-Aparicio Rosa Mar´ıa, Navarro´ Oca˜ na Armando, Manjarrez-Hern´andez H. Angel, Gavilanes-Parra Sandra, AparicioOzores Gerardo, Cauich-S´anchez Patricia Isidra, Garza-Ramos Ulises, Molina-L´opez Jos´e, Characterization of Escherichia coli causing community acquired urinary tract infections in Mexico City, Diagnostic Microbiology and Infectious Disease (2016), doi: 10.1016/j.diagmicrobio.2016.11.006
This is a PDF file of an unedited manuscript that has been accepted for publication. As a service to our customers we are providing this early version of the manuscript. The manuscript will undergo copyediting, typesetting, and review of the resulting proof before it is published in its final form. Please note that during the production process errors may be discovered which could affect the content, and all legal disclaimers that apply to the journal pertain.
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Characterization of Escherichia coli causing community acquired urinary tract infections in Mexico City
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Laura Belmont-Monroya,b, Rosa María Ribas-Aparicioa, Armando NavarroOcañac, H. Ángel Manjarrez-Hernándezc,d, Sandra Gavilanes-Parrac,d, Gerardo Aparicio-Ozoresa, Patricia Isidra Cauich-Sáncheza, Ulises Garza-Ramose, José Molina-Lópezb,c,* a
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Departamento de Microbiología, Escuela Nacional de Ciencias Biológicas, Instituto Politécnico Nacional (IPN). Mexico City, Mexico. b Laboratorio de Patogenicidad Bacteriana, Unidad de Hemato-Oncología e Investigación, Hospital Infantil de México “Federico Gómez”. Mexico City, Mexico. c Departamento de Salud Pública, Facultad de Medicina, Universidad Nacional Autónoma de México (UNAM). Mexico City, Mexico. d Dirección de Investigación, Hospital General Dr. Manuel Gea González, Secretaría de Salud. Mexico City, Mexico. e Instituto Nacional de Salud Pública (INSP), Centro de Investigación Sobre Enfermedades Infecciosas (CISEI), Departamento de Diagnóstico Epidemiológico. Cuernavaca, Mor., Mexico. Running title: E. coli O25-ST131 clone from outpatients, UTI
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*Corresponding author: Departamento de Salud Pública, Facultad de Medicina, UNAM. México D.F. 04510, México. Phone: (52-55) 52 28 99 17. E-mail: [email protected]
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Abstract
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The O25-ST131 clone was identified within 169 uropathogenic Escherichia coli (UPEC) strains. The 44.8% of the 29 O25-ST131 clones detected were positive to least to one
SC
extended-spectrum β-lactamase gene. The phylogroup D was mainly found. The O25-
NU
ST131 clone appeared to be associated with community-acquired UTI in Mexico City.
AC
CE
PT
ED
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Keywords: Urinary infection; Escherichia coli; Multiplex PCR; Serogroups; ST131 clone
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Escherichia coli clone O25-ST131 has emerged worldwide as important cause of community-onset urinary tract infection (UTI) (Nicholas-Chanoine et al.,
RI P
T
2008; Coque et al., 2008). This clone has been often associated with extendedspectrum -lactamases (ESBL) enzymes and is frequently Multidrug-Resistant
SC
(MDR) (Johnson et al., 2002; Rogers et al. 2011). There is no comprehensive surveillance of E. coli causing community-acquired UTI in Mexico City; therefore, it
NU
is very difficult to estimate the true incidence of UTI and to monitor the spread of
MA
this organism. The aim of this study was to determine the O-serogroup of uropathogenic E. coli (UPEC) strains, the prevalence of the clone O25-ST131 in E.
ED
coli isolates from outpatients diagnosed with an UTI, and their association with
PT
phylogroups and the presence of ESBL genes. One hundred sixty nine E. coli isolates, one isolate per patient, were
CE
collected from community-acquired UTI in Mexico. These isolates that met the
AC
inclusion criteria were selected from 800 outpatients and collected between February 2010 and October 2012, at Mexico General Hospital. The outpatient’s aged was between 18 and 70 years. For this study was consider a UTI acquired in the community that was present in apparently healthy patients who only attended the consultation by the onset of symptoms. Excluding criteria were: patients with complicated-UTI; if they were pregnant; if they had diabetes, if they were immunocompromised; or if other suspected sources of infection were present. Identification of isolates was carried out using standard microbiological techniques (Carridge et al., 1988; Devine et al., 1989). An initial serotyping as agglutination
3
ACCEPTED MANUSCRIPT 4
assay (Ørskov and Ørskov, 1984) was carried out, using 96-well microtiter plates and rabbit serum, obtained against 185 somatic antigens. Isolates that failed to
RI P
T
agglutinate with any of these antisera were defined as Non-Typeable (NT), whereas those that did agglutinate with saline solution were termed Auto-
SC
Agglutinable (AA).
NU
Fourteen pairs of primers were used by determination of UPEC serogroups by multiplex PCR, among AA and NT strains detected by conventional serotyping.
MA
Multiplex PCR was performed according to Li et al. (2010). The phylogroups were determined by PCR assay, using a combination of three DNA markers (chuA, yjaA,
ED
and TspE4C2.1) (Clermont et al., 2000). All O25 serogroup isolates were tested to
PT
determine those of the O25-ST131 clone by multiplex PCR assay (allele-specific PCR for the pabB gene) according to Clermont et al. (2009). Finally, both Narrow-
CE
Spectrum -Lactamases (NSBL) and ESBL: TEM-1, CMY-, OXA-1, SHV- and
al. (2013).
AC
CTX-M-15 type were screened by PCR using specific primers according to Kiiru et
Of the 169 E. coli isolates, 142 (84.0%) were typeable, 12 (7.1%) were NT, and 15 (8.9%) were AA strains. Of the typeable strains, 80/142 (47.3%) belonged to UPEC serogroups and 62/142 (36.7%), to non-UPEC serogroups. The O25 serogroup was the most common with 29 (17.1%) of the isolates, followed by O1 (10%), O8 (9%), O6 (4%), and O75 (3%). The NT isolates were identified as O15, O18, O21, and O25 serogroups and AA isolates were identified as O25, O21, O18,
4
ACCEPTED MANUSCRIPT 5
and O8 serogroups. Among the O25 serogroup all corresponded to O25-ST131
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clone (28 identified by conventional serotyping and one by multiplex PCR).
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Otherwise, 64 (37.9%) of the isolates belonged to phylogroup D, 47 (27.8%) belong to group A, 30 (17.8%) to group B1, and 28 (16.5%) to group B2. We found
SC
statistically significant differences between phylogroup B2 and UPEC vs. non-
NU
UPEC serogroups (p value, 0.03), (Table 1). On the other hand, the majority of NT and AA isolates belonged to the A and B2 phylogroups. Twelve (41.4%) of the
MA
O25-ST131 clone identified corresponded to the D phylogenetic group, ten (34.4%)
ED
to B2, four (13.8%) to A, and three (10.4%) to the B1 phylogroup. Thirteen (44.8%) of the 29 O25-ST131 clones detected were positive to
PT
least to one NSBL- or ESBL-type, showed various combinations of -lactamase
CE
genes (Table 2).
The ST131 E. coli isolates are considered to be truly pathogenic, due to the
AC
spectrum of infections that they cause in both community and hospital settings and to the large number of virulence-associated genes that they contain (NicolasChanoine et al., 2014). We found that these strains cause community-acquired UTI in patients in Mexico City; including some E. coli isolates belonging to the O25ST131 clone. In this study, 29 isolates with O25 serogroup were positives to O25ST131. It has been reported that the majority of O25-ST131 isolates are MDR and that they commonly produce CTX-M-15 ESBL enzyme (Nicholas-Chanoine et al., 2008), causing UTI and significant morbidity and mortality; this clone constitutes a
5
ACCEPTED MANUSCRIPT 6
major public health concern (Nicholas-Chanoine et al., 2008; Clermont et al., 2009;
T
Peirano and Pitout, 2010).
RI P
In this study, phylogenetic groups A and D were common (27.8 and 37.9%, respectively) among our isolates. Lopez-Banda et al., 2013 found a high
SC
percentage of strains belonging to the phylogenetic group A (30.6%) and B2
NU
(55.6%) and less frequently B1 (9.3%) and D (4.6%). This was not expected because extra-intestinal virulent strains belong mainly to groups D and B2; we
MA
obtained 17.8% of group-B1 strains and 16.5% of group-B2 strains. Due to that a high percentage of strains of this study belong to group A, it is necessary to
ED
continue to monitor the phylogroups considered as commensal, as in the case of
as
commensal
PT
group A, because these are capable of causing UTI in the population. Thus, its role should
be
reconsidered.
This
result
suggests
that
the
AC
the urinary tract.
CE
gastrointestinal tract is the main reservoir of strains that may be able to colonize
Recently, the E. coli ST131 lineage and phylogenetic group B2 were shown as the major clones among CTX-M-15-producing E. coli isolates obtained from several clinical sources, mainly urine and blood in Australia, Canada, India, Spain, the U.K., and New Zealand. (Petty et al., 2014). In Mexico, in strains from community acquired UTI, it was found that 13/17 isolates with serogroup O25 belonged to the O25-ST131 clone (Molina-López et al., 2011), while Reyna-Flores et al. (2013) found that 25% of 4,735 UPEC isolates belonged to the O25b-ST131
6
ACCEPTED MANUSCRIPT 7
clone, compared to the 17% identified in this study. These reports document the spread and success of the ST131 E. coli lineage. In our study, a low percentage of
RI P
T
ESBL was obtained as the strains of ST131 cloned, and only two strains were positive for the blaCTX-M-15 gene. The O25-ST131 strains found were not associated
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with the presence of ESBL; therefore, we suggest that the resistance of these strains is not mediated by these enzymes. Likewise, we demonstrated the
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prevalence of serogroup O25 strains, which aids in detecting the spread of specific
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clones, such as O25-ST131, among outpatients with UTI in Mexico City.
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Acknowledgements
This study was supported by CONACyT, through its support to project No. 166004,
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Pública, UNAM.
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and PAPIIT number IN228916, Facultad de Medicina, Departamento de Salud
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References
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Nicolas-Chanoine, M.-H., Bertrand, X., & Madec, J.-Y. (2014). Escherichia coli ST131, an Intriguing Clonal Group. Clinical Microbiology Reviews, 27(3), 543–574.
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ST131: A pandemic, multiresistant, community-associated strain. J Antimicrob
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Table 1. Phylogenetic distribution of E. coli isolates by serogroups determined by conventional serotyping and polymerase chain reaction (PCR) P
UPEC serogroups
Non-UPEC
group
(n = 80)
serogroups (n = 62)
value
A
26
21
0.11
B1
16
B2
23
D
36
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0.79
5
0.03
28
0.98
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Phylogenetic
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UPEC = Uropathogenic E. coli
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Table 2. NSBL- and ESBL-genes combination present in E. coli O25-ST131
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isolates from outpatients with UTI.
NSBL- and ESBL genes
strains
combination
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Frequency in O25-ST131
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N (%) TEM-1a
2 (6.7)
TEM-1a/OXA-1a
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7 (24.1)
OXA-1a
1 (3.5)
CTX-M-15b
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1 (3.5)
CTX-M-15b/TEM-1a
1 (3.5)
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16 (55.2)
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1 (3.5)
CMY Negative to -lactamase genes
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NSBL: Narrow-Spectrum -Lactamase
a
ESBL: Extended-Spectrum -Lactamase
b
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Highlights
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The O25-ST131 strains found were not associated with the presence of ESBL.
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The E. coli O25-ST131 isolates negative for -lactamase genes were higher in
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comparison with the presence of NSBL or ESBL genes
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The prevalence of serogroup O25 strains, aids in detecting the spread the O25-
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ST131 clone among outpatients with UTI in Mexico City.
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