2023.08.02.66
Files > Volume 8 > Vol 8 No 2 2023

Molecular detection of Escherichia coli efflux pumps genes isolated
from UTI in pregnant women

Wedad
Adil Kadhim 1,*, Kareem Ibrahim Mubarak 2,*
1 Diyala University/College of Science/Department of Biology
2 Diyala University/College of Science/Department of Biology
* Correspondence: [email protected],
[email protected]
Available
from: http://dx.doi.org/10.21931/RB/2023.08.02.66
ABSTRACT
Sixty-three clinical samples from midstream urine samples were collected
from pregnant women with urinary tract infections. After microscopic
examination, culture and biochemical tests and the final diagnosis using the
VITEK-2 system, 25 Escherichia coli isolates were discovered. Antimicrobial
susceptibility results showed that E.coli isolates were resistant
to gentamicin (%92), amikacin (%28), norfloxacin (%52), Ciprofloxacin (%56),
ofloxacin (%60), trimethoprim (%8), chloramphenicol (%80), colistin sulfate
(%20), tetracyclin (%68), azithromycin (%48), cefoxitin (%40),
amoxicillin-clavulanate (%96), ampicillin (%92). The prevalence of capsule
posses, hemolysin production, biofilm formation, and efflux pumps
wrer%24,%16,%72 and %44 respectively. The result of efflux pumps genes acrA and
acrB gene detection was 100. The acrA and acrB gene expression increased after
treatment with the antibiotic Ciprofloxacin. Because of the primary role of Escherichia
coli in urinary tract infection and the presence of a high ratio of
Multidrug Resistance ( MDR ), and the importance of The efflux pumps in
antibiotics resistance, the current study was conducted to determine the MDR
isolates from UTI in pregnant women's in Baquba city, the percent of acrA and
acrB genes among strains and the effect of Ciprofloxacin treatment on gene
expression.
Keywords. Escherichia coli, efflux pumps, acrA gene, acrB
gene
INTRODUCTION
Escherichia coli is a
gram-negative bacteria, rod-shaped, motile or non-motile, facultatively
anaerobic, lactose fermentative1. It is one of the most common types
of the Enterobacteriaceae family that naturally inhabit the human gut, making
it able to infect other body systems opportunistically when the opportunity
arises and cause many diseases 2. E. coli is the leading
cause of 90% of urinary tract infections, especially in women, and it comes in
second place after respiratory diseases 3. The excessive use of
antimicrobial agents led to the problematic treatment of UTI infection due to
bacteria having the character of multidrug resistance (MDR) as they can produce
many enzymes, such as ESBL Extended Spectrum B-lactamase Enzymes, Metallo
B-lactamase MBL and hemolysin 4
bacteria posses other mechanisms that enable them to resist antibiotic, such as
changing the target site, changing the permeability of the cellular membrane,
inhibiting protein synthesis and efflux pumps. It confers the bacteria
multi-resistance to antibiotic 5.
Efflux pumps are transporters of protein
located in the cell membrane of bacteria and play a significant role in
transporting various substances and expelling them outside the cell to
eliminate their harmful effects. It is an essential way for bacteria to develop
resistance to antibiotics6. These pumps fall into two main
categories based on their genetic origin, which are chromosomal efflux pumps,
that is, they carry the genes that code for them on a cell's chromosome, and
plasmid efflux pumps carry the genes they encode on portable genetic materials,
such as plasmids or transposons 7. There are five types of families
of efflux pumps; Major Facilitator Super Family (MFS), Small Multidrug
Resistance Family (SMR), Family (Multidrug and Toxic Efflux Family (MATE), ATP
- Binding Cassette Family (ABC, Resistance - Nodulation - Division Family (RND)8.
The RND family is classified into three classes according to its components,
which include: Single-component efflux pumps represented by the protein AcrB
located in the inner membrane of the cell, which is encoded by a gene called
acrB, which transports hydrophilic antigens. Two-component pumps represented by
AcrB in the inner membrane and AcrA lipoproteins in the periplasmic space,
encoded by a gene called Acra, Three-component tripartite pumps represented by
AcrB in the inner membrane, AcrA proteins in the plasma vacuole, and a
funnel-like protein channel called ToIC encoded by the tolC gene), which is
located in the outer membrane of a bacterium 9.
Gene
expression is known to be an essential feature for maintaining the functional
integrity of cells. It occurs in different ways and an organized manner and is
divided into positive and negative organizations. Regulation occurs when mRNA
begins to be transcribed in prokaryotic mostly. While it becomes more complex
when it is present in eukaryotes, so there is more than one mechanism to
complete the process of regulation; this process occurs through the association
of proteins with a specific sequence on the DNA strand, resulting in either an
increase or decrease in the transcription rate in eukaryotic and prokaryotic.
In bacteria, this process occurs through the initiation of cloning. An example
is the positive and negative control of the lactose operon system operator in E.
coli 10.
MATERIALS AND METHODS
Collection of Specimens
Sixty-three urine specimens were collected from
pregnant women have symptoms of urinary tract infection admitted to Al-Batool Maternity and Children Teaching
Hospital in Baquba City during the period from 1/9/2021- to 1/12/2021.by using
disposable collection sterilized container to isolate and identify Escherichia
coli from urine specimens and report the information about each patient.
Isolation and identification of bacteria
The urine specimens were inoculated on the
MacConkey agar medium after 24 hours plates were inoculated aerobically
overnight at 37C֯. According to morphological ( shape and grams stain )and
cultural colony characters, the expected isolates were subculture on the Eosine
Methylene Blue (EMB) agar, the confirmation of bacterium identification done by
biochemical tests including (oxidase, catalase
IMVIC tests, urease, Triple sugar
iron). Finally, identification is complete by depending on the VITEK-2
compact system 11.
Antibiotic
sensitivity test and MIC Determination
The susceptibility E.coli isolates to antibiotics
were performed by using 13 antibiotics (gentamicin, amikacin, norfloxacin, Ciprofloxacin,
ofloxacin, trimethoprim, chloramphenicol, colistin sulfate, tetracyclin,
azithromycin, cefoxitin, amoxicillin-clavulanate, ampicillin) the base of the Disk Diffusion method by
spreading bacteria on the Muller-Hinton Agar medium (Vandepitte et al.;2003) the interpretation
of results and classify isolates as sensitive and resistance by ( CLSI 2019 )—determination of MIC of ( Ciprofloxacin
& Norfloxacin) 12.
Detection of
Virulence Factors of E.coli
Several virulence factors of Escherichia coli were
detected in bacterial strains, including
Hemolysin production
Bacterial isolates were cultured on plates
containing blood agar medium, and the plates were incubated at 37 °C for 22 hours;
then, the results were read by observing the type of hemolysis zones around the
bacterial colonies13.
Detection of Capsule
The Indian ink method was used for dyeing; take a colony
of bacteria and mixed with a small drop of physiological saline using a wooden
stick on a clean glass slide; add a drop of Indian ink to the slide and mix,
then the slide cover was placed over it so that the liquid spread in a light
layer under the glass cover, and examined under the oil lens14.
Detection of Biofilm formation
Phenotypic detection of Biofilm formation, Micro Titer Plate assay
according to 15 to detect the biofilm formation, bacteria were inoculated on Nutrient broth
medium and incubated at 37°C for 24 hours. After that, the broth cultures were
compared with a McFarland Standard using the same medium as were transfer 200 μl of an isolate suspension into each of
three wells of a 96-well flat-bottomed polystyrene plate and incubated at37°C
for 24 hours. Each well was washed thrice with distilled water (DW) with rough
shaking and dried thoroughly. Absolute methanol(200μl) was used for fixation of
adhering bacterium. After that, each well was stained with 200μl of 0.5%
Crystal Violet for 15 minutes. .washing several times was.to remove the excess
stain. Later, the crystal violet associated with adherent cells was retained
with 200μl of ethanol per well. The absorbance of wells filled with
bacteria-free Nutrient broth served as a negative control. The amount of
crystal violet removed by 95% ethanol in
each well was quantified by measuring the OD 630 using an ELISA reader.
Because. of this, the absorbance values. Represented the intensity of the
biofilm formed by well-studied isolates on the surface of the microtiter. The
results obtained were categorized into three groups (i.e., non-biofilm
producer, moderately and firmly).
Where the absorption of the cultivated pit was
compared with the control pits as follows: If OD≤ ODc (Considered non-biofilm
producer); if ODc ≤ OD ≤2*ODc (Considered moderately biofilm producer); if
2*ODc ≤ OD( Considered strongly biofilm producer). Where OD (Represent the
tested isolates); ODc (Represent control pits).
Phenotypic Efflux pumps detection
Phenotypic Detection of Efflux pumps was conducted by(cartwheel
pattern) agar-EtBr method(Martins et al.;2013); dilutions of bacterial were
prepared from isolates that revealed the resistance to antibiotics and the
ability to biofilm formation after incubated for 24 hours at 24C֯ in the 5 ml of buffer. Their concentration was adjusted by
comparing to 0.5 of MacFarland standard solution, then isolates inoculated on
tryptic soy agar (TSA) plates containing(0,5,10,15,20,25 µg\ml ) of Ethidium
Bromide(EB), the plate was divided to 16 by radial lines(cartwheel pattern), the
plates incubated for 16 hours at 37 C֯ in the dark place, the isolates were examined under ultraviolet
(UV) trans illuminator to record the fluorescent.
Genotypic detection of efflux pumps
DNA extraction: DNA bacteria was extracted from the
isolates of E.coli by using a genomic DNA purification Kit supplemented by the manufactured
company (Geneaid, Thailand)16.
Polymerase Chain Reaction (PCR) detected acrAB
efflux pump genes (acrA,acrB)in E.coli isolates. The proliferation of acrAB
genes with two primer pairs(Bioneer, Korea) Table 1. The amplification of DNA
was performed at a volume of 25µl; the reaction mixture contained 5µlAccu
powerPCRPremix (Bioneer, Korea), 3µl template DNA, 1.5µl F-primer,1.5µl
R-primer and 14µl deionized nuclease-free water. PCR reaction of acrAB genes was
performed according to 8 in three steps in Table 2.

Table
1. Sequences and primers concentration used in the study

Table
2. Phases of the polymerase chain reaction
in the thermal cycler
Gel electrophoresis: after the amplification process in the PCR
apparatus, the product was run on 1.5 % agarose gel with 5ml of ethidium
bromide in 1x (TBE) buffer using a DNA ladder (100-2000)bp (Bioneer, Korea) at
100 volts for 80 minutes. The visualization of PCR products under 320 nm UV
light using a UV transilluminator 12.
Ciprofloxacin Effect on gene expression
RNA Isolation
E.coli Bacteria was pelleted and then lysed in 1 ml of AccuZolTM lysate through the. Two hundred
µl of chloroform was added per 1ml of AccuZolTM and shaken vigorously for 15
seconds. The mixture was incubated on ice for 5 minutes. The mixture was
centrifuged at 12000 rpm for 15 minutes at 4C, The aqueous phase was
transferred to a new 1.5ml tube, and an equal volume of isopropyl alcohol was
added. The tubes were mixed by inverting 4 – 5 times and incubated at -20C for
10 minutes. Then Centrifuge at 12000 rpm for 10 minutes at 4C was done, and the
supernatant was carefully removed. One ml of 80% ethanol was added and mixed well
by vortexing and Centrifuge at 12000 rpm
for 5 minutes at 4C was done, then the supernatant was carefully removed, and
The pellet was dried; the RNA was dissolved in RNase – free water by passing
the solution a few times through the tip of the pipette and stored at -80%
until us 18.
Transformation of RNA to DNA
The RNA extracted from each bacterial isolate, primer and water were
mixed and incubated at 65°C for 5 minutes, on ice for two minutes. RNA was
10pg-500mg, random primer 1µl, 2XES reaction mix 10µl, RT\RL enzyme 1µl, gDNA
remover 1µl, RNase- free water 20µl (Bioneer, Korea). The mixture was treated
with 25c֯ for 10 minutes,42c֯ for 15 minutes, and 85c֯ for 5 seconds to inactive enzymes.
Real-time quantitive reverse transcription (qRT-PCR)
RT-qPCR is performed in a reaction solution of 20µL volume using
BrightGreen qPCR MasterMix by mixing the
0.6 µL of primer, 10µL of BrightGreen 2X qPCR master mix, ≤ 500 ng of
cDNA, Nuclease-free H2O to 20 µl.
The generated solution was placed in Real-time PCR Cycler for thermal
reaction to measure the Cycle Threshold (CT) value. RT-PCR is used for
quantification of the levels of gene expression. The measured CT values during the
thermal reaction are recorded to the computer in the following measurements 19.
Real-Time qRT-PCR analysis: ΔCT (test) = CT gene of interest (target,
test) – CT internal control, ΔΔCT=ΔCT (test)- ΔCT (calibrator), 2-ΔΔCt
= Normalized expression ratio(20).
Statistical analysis
Statistical analysis was carried out using the SPSS version 20, and the
results were analyzed using the Chi-Square test with a probability level of P≥
0.05 21.
RESULTS
The diagnosis results showed that the number of E.coli isolates obtained
from a total of 63 urine specimens was 25. Biochemical tests reveal that all
the isolates of E.coli were positive for catalase, Indole, and Methyl Red and
negative for Oxidase, Vogues-Proskauer, citrate utilization and Urease, TSI(A\A,
H2S+, gas+).E.coli isolates were gave pink colonies on MacConkey agar, dark
blue with green metallic sheen on(EMB)agar.
Sensitivity of E.coli to Antibiotic
The results of the current study showed the resistance ratio to
Gentamicin, Tetracyclin, and Trimethoprim were 23(%92),17(%68), and 2(%8), respectively.

Table 3. Sensitivity
of Escherichia coli isolates to antibiotics.
MIC
Determination
The
current study revealed a contrast in the MIC values of E.coli
isolates against the antibiotics ciprofloxacin and norfloxacin for isolates of
pregnant women infected with urinary tract infections. Where it ranged between
(8-1024) µg/ml for Ciprofloxacin and (8-512) µg/ml for norfloxacin, and the
range of MIC for the four isolates (2, 5, 30, 24) were treated to study the
gene expression reach(8-256)µg\ml.
Hemolysin
production: The results of the current study showed that 4(%16) of E.coli
produced hemolysin.
Capsule
posses: The results of the current study showed that 6 (%24) of E.coli
possess a capsule. This result approached 26, who found that %37.2
of E.coli isolates are capsule posses.
Biofilm
formation: efflux pumps production: The results of the current study showed that
11(%44) of E.coli produced efflux pumps.
Detection
of (acrAB)genes: molecular detection indicates that all isolates
carrying acrAB genes by %100 were carriers Figure 1,2.

Figure 1. Electrophoresis of amplified acrA(105bp) for E.coli
by 1.5% agarose gel, using DNA Ladder 100-2000bp, 80V\cm for 80 min, stained
with ethidium bromide dye and visualized under UV transilluminator
documentation All lanes are shown to have the gene.

Figure 2.
Electrophoresis of amplified acrB (107bp) for E.coli by
1.5% agarose gel, using DNA Ladder 100-2000bp, 80V\cm for 80 min, stained with
ethidium bromide dye and visualized under UV transilluminator documentation All
lanes are shown to have the gene.
Ciprofloxacin
Effect on Gene Expression
The
results of the current study for four bacterial E.coli of
isolates (2, 5, 30, 24) isolated from pregnant women before and after treatment
with Ciprofloxacin using the qRT-PCR technique. Using the primers described in
Table 1. showed amplification of the acrA gene with a value of threshold
cycle(CT ) before isolates (2,5,30,24) treated with sub-MIC CIP, Where recorded
(31.62, 30.42, 27.79, 28.99). The quantitative changes in mRNA were determined
using an equation Threshold limit Ct (2-△△ct), as shown in Table 4, after treatment of
the isolates with CIP. Isolates (2, 5) showed high genetic expression after
treatment with cip where recorded (3.9737, 9.0630). As for the (30, 24), It was
not genetically expressed after treatment with cip Table 4.

Table 4.
Effect of ciprofloxacin treatment on CT
and Fold values of acrA gene expression for E.coli
Results
of the current study for four bacterial E.coli of isolates (2, 5,
30, 24) isolated from pregnant women before and after treatment with
Ciprofloxacin using qRT-PCR technique. Using the primers described in Table 1.
showed amplification of the acrB gene with a value of threshold cycle(CT
) before isolates (2,5,30,24) treated with sub-MIC CIP, Where recorded (22.75,
22.65, 24.7, 28.3) the quantitative changes in mRNA were determined using an
equation Threshold limit Ct (2-△△ct), as shown in Table 5. After treatment of
the isolates with CIP. Isolates (2, 5) showed high genetic expression after
treatment with cip where recorded (5.3889,4.3771). As for the (30, 24), It was
not genetically expressed after treatment with cip Table 5. Our study agreed with
(Ma et al;1995), which showed increased gene expression for acrAB
after treatment with cip.

Table 5. Effect
of ciprofloxacin treatment on CT and
Fold values of acrB gene expression for E.coli
DISCUSSION
The results of the current study showed the
resistance ratio to Gentamicin, Tetracyclin; Trimethoprim was 23(%92),17(%68),
and 2(%8), respectively. This result did not agree with Jazayeri 22,
which showed that the resistance to the antibiotics were%70,%30,%40
respectively, while the resistance ratio of E.coli isolates to Amoxicillin-Clavulanate was 24(%96). This
result was similar to 22, which showed that the resistance to the
antibiotics were%96, while the resistance ratio of E.coli isolates to Ciprofloxacin was 14(%56). This result was
similar to 23. the resistance ratio of E.coli isolates to Ampicillin was 23(%92). This result did not
agree with 23, which showed that the resistance to the antibiotic %
was 81.7. the resistance ratio of E.coli isolates to chloramphenicol and
norfloxacin were20 (%80). this result did
not agree with 24. which showed that the resistance to the
antibiotics were%42.whil the resistance ratio E.coli isolates to Amikacin was 7(%26).this result was approached
to 24. which showed that the resistance to the antibiotic was
%21.whil the resistance ratio E.coli isolates to Cefoxitin10 (%40). This result
was approached to 25, showing that the antibiotic resistance was 42%.
The resistance ratio E.coli isolates to ofloxacin15 (%60). Another study does
not agree with the current study, where resistance to the antibiotic was %39.6
25. The current study showed that
4(%16) of E.coli produced hemolysin. This result does not agree with 24,
who found that %41.9 of E.coli isolates are hemolysin-producing. The current
study showed that 18 (%72) of E.coli formated biofilm. This result approach 27,
who found that %90 of E.coli isolates are biofilm formation. The results of the
current study showed that 11(%44) of E.coli produced efflux pumps. This result
does not agree with 27, who found that %70 of E.coli isolates are
efflux pump production. Moreover, molecular detection indicated that all
isolates carrying acrAB genes by %100 were carriers; the results agreed with
the results of 27.
CONCLUSIONS
The results of isolation and diagnosis of Escherichia coli bacteria
showed multi-resistance to antibiotics due to their possession of efflux pumps.
Moreover, after treatment with ciprofloxacin antibiotic, the results of the
gene expression of acrA and acrB genes recorded high expression in pregnant
women.
REFERENCES
1.
Jaweetz E, Melnick J A and Adelberg
E A. Review of Medical Microbiology 27th ed. McGraw-Hill Higher Education. 2016;
pp 850.
2.
Nascimento, J. A., Santos, F. F.,
Valiatti, T. B., Santos-Neto, J. F., M Santos,A. C., Cayô, R., ... & AT
Gomes, T. Frequency and Diversity of Hybrid Escherichia coli Strains Isolated
from Urinary TractInfections. Microorganisms, 2021; 9(4),
693. 13.
3.
ALobaidy, B. .; Al-Falahi, M. N. .;
Almarie, A. A. . Effect Of 6-Bap Growth Regulator On Seed Priming Of Several
Bread Wheat Varieties Under Water Irrigation Salinity Stress. JLSAR
2021, 2, 21-26..
4.
Ye, Q., Wu, Q., Zhang, S., Zhang,
J., Yang, G., Wang, H., ... & Wang, J. antibiotic-resistant extended
spectrum ss-lactamase-and plasmid-mediated AmpC-producing Enterobacteriaceae
isolated from retail food products and the pearl river in Guangzhou, China. Frontiers
in microbiology, 2017; 8, 96. 17.
5.
Kapoor, J.; Saigal, S. and
Elongavan, A. Action and Resistance Mechanisms of Antibiotics: A Guide for
Clinicians. J Anaesthesiol Clin Pharmacol. 2017; 33(3):300-305.
8.
6.
Zhi Li, X.; Elkins, C. A. and
Zgurskaya, H. I. Efflux-Mediated Antimicrobial Resistance in Bacteria
Mechanisms, Regulation And Clinical Implications. International Publishing.
Switzerland. 2016; 848 pp.
7.
Puzari, M. and Chetia, P. RND
efflux pump mediated antibiotic resistance in GramNegative Bacteria Escherichia
coli and Pseudomonas aeruginosa: A Major Issue Worldwide. World J Microbiol
Biotechnol. 2017; 33(24): 1-8. 14.
8.
Maleki, D.; Jahromy, S. H.; Karizi,
S. Z. and Eslami, P. The prevalence of acrA and acrB Genes Among Multiple-Drug
Resistant Uropathogenic Escherichia coli Isolated from Patients with UTI in
Milad Hospital, Tehran. Avicenna J Clin Microb Infec. 2017; 4(1):
1-7. M
9.
Hayashi, K.; Nakashima, R.;
Sakurai, K.; Kitagawa, K.; Yamasaki, S.; Nishino, K. and Yamaguchi, A.
AcrB-AcrA Fusion Proteins That Act as Multidrug Efflux Transporters. J
Bacteriol. 2016; 198(2):332–342. 7.
10. Maysaloon
W. Ibraheem, Abdulkhaliq A. Farhan, Sataa M. Salih, Th.T. Mohammed. Carcass
characteristics of Awwasi lambs supplemented with Selenium and Vitamin D3. Iranian
Journal of Ichthyology, 2022; 9(1) : 355-359.
11. Wanger
A, Chavez V, Huang R S P, Wahed A, Actor J K and Dasgupta A. Microbiology And
Molecular Diagnosis In Pathology. Elsevier Inc.All Rights Reserved. 2017;
300pp.
12. Moerllo
, J.A. ; Granto , PA and Mizer , H.E. LaboratoryManual and workbook in
Microbiology : Applications to patient Care. 2003.
13. Tille,
P.M. Baily And Scott's Diagnostic Microbiology. 41thed. Elsevier,Inc. China. 2017;
1115pp.
14. Atlas,
R. M. ; Brown, A. E. and Parks, L. C. Laboratory Manual Experimental
Microbiology. 1st. ed. Yearbook , Mosby Inc. 1995;148: 884-8.
15. Fusco,
A.; Coretti, L.; Savio, V.; Buommino, E.; Lembo, F. and Donnarumma, G. Biofilm
Formation and Immunomodulatory Activity of Proteus mirabilis Clinically
Isolated Strains. Int J Mol Sci. 2017;18(2): 1-11.
16. Sambrook
J and Russell D. Molecular Cloning Laboratory Manual. 3rd ed. Cold Spring
Harbor, New York. USA. 2001.
17. Lee,P.Y.,Costumbrado,J.,Hsu,C.Y.,&Kim,Y.H.
Agarose gel electrophoresis for the separation of DNA fragments . Journal of
visualized experiments:JoVE, 2012 ; (62).1-6.
18. Chomczynski,
P., & Sacchi, N. The single-step method of RNA isolation by acid
guanidinium thiocyanate–phenol–chloroform extraction: twenty-something years on.
Nature protocols, 2006; 1(2), 581- 585.
19. Abtan,
J., Bhatt, D. L., Elbez, Y., Sorbets, E., Eagle, K., Reid, C. M., ... &
REACH Registry Investigators. Geographic variation and risk factors for
systemic and limb ischemic events in patients with symptomatic peripheral
artery disease: Insights from the REACH Registry. Clinical cardiology, 2017;
40(9), 710-718.
20. Livak,
K. J., & Schmittgen, T. D. Analysis of relative gene expression data using
real-time quantitative PCR and the 2− ΔΔCT method. methods, 2001;
25(4), 402-408. 10.
21. Levesque
R. SPSS Programming and Data Management. 4th ed. Chicago. 522. 2007.
22. ,
M. A., & IRAJIAN, G. R. Asymptomatic urinary tract infection in pregnant
women. 2009.
23. Demilie,
T., Beyene, G., Melaku, S., & Tsegaye, W. Urinary bacterial profile and
antibiotic susceptibility pattern among pregnant women in North West Ethiopia. Ethiopian
journal of health sciences, 2012; 22(2).
24. Mishra,
S., Mohapatra, S., Ranjit, M. R., & Panda, G. K. Biofilm Formation as a
Uropathogenic Marker of E. Coli Isolates Causing Urinary Tract Infections among
Pregnant Women.
25. Thapa,
R., Lamichhane, P., Banjara, M. R., & Acharya, G. P. Prevalence Of Extended
Spectrum Beta Lactamase Producing Uropathogens In Pregnant Women. Prevalence,
8(1). Jazayeri, M. A., & Irajian, G. R. (2009). Asymptomatic
Urinary Tract Infection In Pregnant Women. 2015.
26. Olorunmola,
F. O., Kolawole, D. O., & Lamikanra, A. Antibiotic resistance and virulence
properties in Escherichia coli strains from cases of urinary tract infections. African
journal of infectious diseases, 2013; 7(1), 1-7.
27. Al-Saadi,
Z. H. A., & Abdullah, R. M. Phenotypic And Molecular Detection Of
Escherichia Coli Efflux Pumps From Uti Patients. Biochemical And Cellular
Archives, 19(Suppl. 1), 2371-2376. 2019.
Received: May 15, 2023/ Accepted: June 10, 2023 / Published:
June 15, 2023
Citation: Kadhim, W.A.; Mubarak, K.I. Molecular detection of
Escherichia coli efflux pumps genes isolated from UTI in pregnant women. Revis Bionatura
2023;8 (2) 66. http://dx.doi.org/10.21931/RB/2023.08.02.66