TO STUDY THE MOLECULAR PROFILING AND ITS ASSOCIATION TO EXTENDED SPECTRUM Β-

LACTAMASE (ESBL) PRODUCING E.COLI ISOLATES WITH SPECIAL REFERENCE TO blaTEM ,

blaCTX AND blaSHV GENE FROM PATIENTS OF URINARY TRACT.
SEEJPH Volume XXVI, S1, 2025, ISSN: 2197-5248; Posted:05-01-2025

TO STUDY THE MOLECULAR PROFILING AND ITS ASSOCIATION TO

EXTENDED SPECTRUM Β-LACTAMASE (ESBL) PRODUCING E.COLI ISOLATES
WITH SPECIAL REFERENCE TO blaTEM , blaCTX AND blaSHV GENE FROM
PATIENTS OF URINARY TRACT.

Dr. Suraiya Khanam Ansari, Dr. Asha G, Dr. Mukesh Kumar, Dr. Nashra Afaq, Dr.
Rajesh Verma*

Associate Professor and Head1 , Department of Microbiology, G.S.V.M. Medical College, Kanpur, Uttar
Pradesh, India.
Assistant Professor2 , Department of Biochemistry, Chikkaballapur Institute of Medical Sciences,
Chikkaballapura, Karnataka, India.
Junior Resident3 , Department of Microbiology, King George Medical University, Lucknow, Uttar
Pradesh, India.
Research Associate4 , Department of Microbiology and Central Research Laboratory, Rama Medical
College Hospital and Research Centre, Kanpur, Uttar Pradesh, India.
Professor and Head * , Department of Microbiology, UPUMS Saifai Medical College, Etawah, India.

Corresponding Author: Dr. Rajesh Verma*

Email ID: [email protected]

KEYWORDS ABSTRACT
ESBL, UTI, MDR, Introduction: Longer hospital stays, higher treatment costs, and fewer
β-Lactams , treatment options—particularly wide-spectrum antibiotics—are often
CLSI,DNA, PCR, linked to the rise of extended-spectrum β-lactamase (ESBL). Among the
blaCTX, blaTEM, most commonly given antibiotics in human medicine are β-lactams.
blaSHV However, resistance to these drugs has increased dramatically due to their
widespread and mostly improper use, especially as a result of the
development of extended-spectrum β-lactamase (ESBL).
Aim and Objective: To study the molecular characterization of extended
spectrum β-lactamase (ESBL) producing E.coli isolates with special
reference to blaTEM, blaCTX and blaSHV gene from patients of urinary
tract.
Material and Methods: This was a Cross sectional study carried out in
the department of Microbiology for a period of 12 months i.e, November
2023 to November 2024. A total of 366 E. coli isolates of all the
Uropathogenic E. coli strain isolated from urine samples collected from
hospitalized and consultation patients were included in the study. The
Antimicrobial susceptibility testing was performed according to the CLSI
guidelines 2024. The DNA extraction was done using the Qiagen DNA
extraction kit and the gene blaTEM, blaCTX and blaSHV was detected
using the PCR.

Results: In the present study out of the 1012 isolates there were 366 (35.5%)

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LACTAMASE (ESBL) PRODUCING E.COLI ISOLATES WITH SPECIAL REFERENCE TO blaTEM ,
blaCTX AND blaSHV GENE FROM PATIENTS OF URINARY TRACT.
SEEJPH Volume XXVI, S1, 2025, ISSN: 2197-5248; Posted:05-01-2025

which showed the growth of E. coli. In which 82 (22.4%) were phenotypically

identified as ESBL producers and 284 (77.6%) were Non-ESBL. Out of the
366 isolates, 110 (30%) were Males and 256 (69.9%) were Females patients.
The overall susceptibility of ESBL isolates to various antibiotics was as
Ampicillin (17.20%), Ampicillin/Sulbactam (28.5%), Gentamycin (65.7%),
Cefoxitin (51.0%), Amikacin (80%), Ciprofloxacin (48%), Meropenem
(97.2%), Ceftazidime(0%), Ceftazidime/ clavunalate(100%),
Piperacillin/tazobactam (85.7%), Ceftriaxone(0%), Nitrofurantoin(100%),
Tigecycillin(97.2%) and fosfomycin(97.2%). In the current study out of the
total 82 isolates there were 42 (51.2%) observed positive for blaTEM gene, 24
(29.2% ) observed positive for blaCTX and 13 (15.8%) observed positive for
blaSHV gene.

Conclusion: There are now fewer treatment choices and increased medical
costs as a result of the substantial expansion of E. coli that produces
ESBL. Due to the significant growth of E. coli that produces ESBL, there
are now fewer options for treatment and higher medical expenses. To
assist clinical care management, efficient infection control, and appropriate
antibiotic treatment, trends for regional epidemiological data on
antimicrobial resistance need to be updated.

INTRODUCTION
Antibiotics are the first drugs of choice to treat infectious diseases. A rise in infectious diseases,
increasing rate of drug resistance, and indiscriminate use of antibiotics are the reasons behind the
high usage of antibiotics in developing countries [1,2]. Antimicrobial resistance (AMR) has a
negative impact on achieving Sustainable Development Goals (SDG), food safety, and food
security. In the antimicrobial resistance (AMR) era, the evolving resistance caused by extended-
spectrum β-lactamases (ESBLs) led to higher morbidity, prolonged hospital stays, and expensive
treatment options [2].
The rise of antimicrobial resistance, particularly from extended-spectrum β-lactamase
producing Enterobacteriaceae (ESBL-E), poses a significant global health challenge as it
frequently causes the failure of empirical antibiotic therapy, leading to morbidity and mortality
[3].
ESBLs are Gram-negative bacteria of the Enterobacteriaceae family that carry ESBL genes in
their plasmids or chromosomes, produce β-lactam hydrolyzing enzymes, and are rightly
considered to be among the most challenging pathogens by the World Health Organization
(WHO) [4]. ESBL-producing Enterobacteriaceae (ESBL-E) confer resistance to penicillin—in
addition to aztreonam and first-, second-, and third-generation cephalosporins—but are unable to
hydrolyze cephamycin or carbapenems [5,6]. Carbapenem has been the drug of first choice for
treating ESBL-E-induced infection for a long time . This is changing, though, due to many
factors including the recent emergence of carbapenemase-producing bacteria. Thus, there is an
urgent need to develop alternative approaches.

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LACTAMASE (ESBL) PRODUCING E.COLI ISOLATES WITH SPECIAL REFERENCE TO blaTEM ,
blaCTX AND blaSHV GENE FROM PATIENTS OF URINARY TRACT.
SEEJPH Volume XXVI, S1, 2025, ISSN: 2197-5248; Posted:05-01-2025

The ESBL-encoding genes are highly diverse in nature and can be classified into many families
with unique characteristics such as blaTEM, blaSHV, and blaCTX-M. TEM 1, the first plasmid
and transposon-mediated β-lactamase, was isolated from the blood culture of a named Temoniera
in Greece in the early 1960s [7] . It has spread worldwide and is now found in many species of
the family Enterobacteriaceae, P. aeruginosa, Hemophilus influenzae, and Neisseria
gonorrhoeae . The SHV-1 type is common in Klebsiella spp. and E. coli . CTX-M-type ESBL
are predominant in E. coli, K. pneumoniae, S. enterica serovar Typhimurium, and Shigella spp.
The plasmid-mediated OXA and AmpC-type ESBL were discovered in P. aeruginosa and K.
pneumoniae isolates, respectively [8].
ESBLs make it difficult to treat infections in acute critical care settings and are most commonly
seen in Enterobacteriaceae in India. In India as well as other countries, CTX-M, TEM, and SHV-
type ESBL are now common, and hospital-associated pathogenic bacterial strains are expressing
more of these enzymes, which could spread widely. ESBLs from a number of nations have also
been described, including OXA-1, PER-type, GES-type, and VEB-type. Using various
techniques to identify ESBL resistance, many researchers in India have reported prevalences
ranging from 7.0 to 91% [9] .
It is important to note that some β-lactamases may not be inactivated by some classical inhibitors
such as clavulanate acid, sulbactam, and tazobactam [10]. Mechanisms of resistance in Gram-
negative bacteria may also involve reduced membrane permeability through genomic mutations,
decreased amounts of β-lactam antibiotics that can enter the cell, and a marked increase in
antibiotic reflux from the periplasm to the exterior of the cell [11]. Massive and usually
inappropriate use of antibiotics for treatment of UTIs generates a selective pressure that is
followed by the rapid emergence and spread of multi-drug resistant bacterial strains. Nowadays,
resistance of uropathogenic E. coli to many antibiotic classes is a very common finding in human
medicine and is usually associated with increased medical costs, prolonged hospital stays and
frequent therapeutic failure [12].
The extended-spectrum β-lactamases, which are currently found all over the world, are the main
class of enzymes used in epidemiology. According to Ambler's molecular and structural
classification system, ESBLs fall under class A.A. Their ability to hydrolyse broad-spectrum β-
lactam antibiotics and their resistance to β-lactamase inhibitors, especially clavulanate, set them
apart biochemically [13].
Compared to traditional phenotypic methods, polymerase chain reaction (PCR)-based molecular
techniques are quick, precise, and offer higher sensitivities for identifying ESBL-resistant genes.
In addition to providing practitioners with a focused treatment strategy, they also aid in epidemic
containment and infection control policy implementation.
Polymerase chain reaction (PCR)-based molecular methods are rapid, accurate, and have better
sensitivities for detecting ESBL-resistant genes than conventional phenotypic methods. They
help clinicians with a targeted approach for the treatment and also help in containment of
outbreaks and the implementation of infection control policies [14].
Moreover, there is an urgent need to develop precise diagnostic tools, new drugs, and novel
strategies against difficult-to-treat antibiotic-resistant pathogens, including the use of antibiotics
in combination or with adjuvants, bacteriophages, antimicrobial peptides, nanoparticles,
antibacterial antibodies, and photodynamic light therapy
Therefore the present study was undertaken to study the molecular characterization of extended
spectrum β-lactamase (ESBL) producing E.coli isolates with special reference to blaTEM , bla
CTX and blaSHV gene from patients of urinary tract.

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LACTAMASE (ESBL) PRODUCING E.COLI ISOLATES WITH SPECIAL REFERENCE TO blaTEM ,
blaCTX AND blaSHV GENE FROM PATIENTS OF URINARY TRACT.
SEEJPH Volume XXVI, S1, 2025, ISSN: 2197-5248; Posted:05-01-2025

MATERIAL AND METHODS

This was a Cross sectional study carried out in the Department of Microbiology for a period of
12 months i.e, November 2023 to November 2024 at a tertiary care centre. A total of 366 E. coli
isolates of all the Uropathogenic E. coli strain isolated from urine samples collected from
hospitalized and consultation patients were included in the study. The Antimicrobial
susceptibility testing was performed according to the CLSI guidelines 2024 [15].
The study population included patients of all age groups and included both male and female were
included
 To detect Extended spectrum beta lactamases in E. coli isolates from urinary tract
infection by phenotypic method combination dics diffusion test.
 To study Antimicrobial susceptibility of novel beta-lactum / beta-lactamases inhibitor
combination drugs (ceftazidime-avibactam).
 To detect bla SHV, blaCTX and bla TEM gene in these multidrugs resistance isolates by
PCR.
PROCESSING IN LABORATORY
All the Urine sample were collected in a clean universal container & processed according to
standard laboratory protocols. In order to confirm UTI, urine samples were also examined under
a microscope, paying special attention to any pus cell presence. One microliter of urine was
inoculated with a medium lacking in cysteine lactose electrolytes deficient agar (CLED; Hi-
Media Laboratories, Mumbai, India). Customary biochemical tests were used to identify the
bacterial culture that flourished under pure culture and in large numbers (>10⁵ cfu/ml for
midstream urine samples). Their antibiotic vulnerability was also assessed in accordance with
CLSI criteria.
ANTIBIOTIC SUSCEPTIBILITY TESTS
The samples were assessed for their vulnerability by disc diffusion technique (DDT) as per the
CLSI guidelines 2024 . The subsequent discs of antibiotics (drug concentrations in µg) were
used: like Ampicillin(10), Ampicillin/ Sulbactam (10/20), Gentamycin (30), Cefoxitin (30),
Amikacin (30), Ciprofloxacin (10), Meropenem (10), Ceftazidime(30), Ceftazidime/
clavunalate(30/10), Piperacillin/ tazobactam(100/10), Ceftriaxone(30), Nitrofurantoin(30),
Tigecycillin(15) and fosfomycin (20).
PHENOTYPIC METHOD FOR DETECTION OF ESBL
Production of ESBL was confirmed with disk diffusion test using 30 µg ceftazidime (CAZ) and
with a combination of 30µg +10µg ceftazidime along with clavulanic acid (CAC) discs (Hi-media,
Mumbai) placed at a distance of 25 mm on a Mueller-Hinton Agar plate incubated by a bacteria
(standard of 0.5 McFarland turbidity) and further kept alive of night long at 37 ◦C. A ≥ 5mm
increase in inhibition diameter diameter of inhibition area for the mixture disc against disc of
ceftazidime establish the systhesis of ESBL. Escherichia coli ATCC 25922 was utilized as
positive ESBL control strain throughout our study.
GENOTYPIC METHOD
MOLECULAR METHODS: For the detection of the gene blaTEM, blaCTX and blaSHV gene the
chromosomal DNA from the clinical strains of E.coli was extracted. The DNA extraction was
carried out using a commercial available DNA extraction kit (Qiagen DNA Extraction Kit) as
indicated by the manufacturer’s instructions.
The extracted DNA was run in PCR for its extension according to standard method.

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LACTAMASE (ESBL) PRODUCING E.COLI ISOLATES WITH SPECIAL REFERENCE TO blaTEM ,
blaCTX AND blaSHV GENE FROM PATIENTS OF URINARY TRACT.
SEEJPH Volume XXVI, S1, 2025, ISSN: 2197-5248; Posted:05-01-2025

Fig No.1 The DNA Extraction Reagents

DNA extraction and PCR method

 Total genomic DNA was extracted from all the ESBL positive isolates using a DNA
extraction kit according to the manufacturer’s instructions. Amplification and detection of
the considered gene was done by the PCR method using specific primers.
 The primers were purchased from “Saha gene’ and was reconstituted with sterile double
distilled water based on the manufacturer’s instruction.

Fig No. 2: Primers for TEM gene Fig No.3: Primers of SHV gene
Polymerase Chain Reaction (PCR)
 The amplification of the blaTEM, blaCTX and blaSHV gene sequence was performed using
PCR.
TARGET GENE PRIMER LENGTH
blaSHV Forward-5; -TTATCTCCCTGTTAGCCACC-3’ 795 [16]
Reverse- 5’ - GATTTGCTGATTTCGCTCGG-3’

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LACTAMASE (ESBL) PRODUCING E.COLI ISOLATES WITH SPECIAL REFERENCE TO blaTEM ,
blaCTX AND blaSHV GENE FROM PATIENTS OF URINARY TRACT.
SEEJPH Volume XXVI, S1, 2025, ISSN: 2197-5248; Posted:05-01-2025

Table 1: Primers used for bla-SHV gene

bla-TEM gene
TARGET GENE PRIMER LENGTH
blaTEM Forward-5’ -ATGAGTATTCAACATTTCCGTG-3’ 861 [16]
Reverse-5’ -TTACCAATGCTTAATCAGTGAG-3’

Table 2: Primers used for bla-TEM gene

bla-CTX gene

TARGET PRIMER LENGTH

GENE
Forward- 5’ -SCSATGTGCAGYACCAGTAA-3’ 544 [16]
blaCTX Reverse-5’ -CCGCRATATGRTTGGTGGTG-3’

Table 3: Primers used for bla-CTX gene

Polymerase Chain Reaction (PCR)

For the PCR amplification, 2 µl of template DNA was added to 18 µl reaction containing 10 µl
of Qiagen master mix, 2 µl of primer mix (1 µl each of the respective forward and reverse
primers) and 6 µl of molecular-grade water.

The cyclic conditions for blaTEM gene, initial denaturation at 95 °C for 15 min, 30 cycles of
94 °C for 30 s, 59 °C for 1 min 30 s and 72 °C for 1 min 30 s were followed by extension of
72 °C for 10 min.
The PCR cycling conditions

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LACTAMASE (ESBL) PRODUCING E.COLI ISOLATES WITH SPECIAL REFERENCE TO blaTEM ,
blaCTX AND blaSHV GENE FROM PATIENTS OF URINARY TRACT.
SEEJPH Volume XXVI, S1, 2025, ISSN: 2197-5248; Posted:05-01-2025

Step
Program
blaTEM Cycles
Time Temperature

Initial denaturation 15 min 95 ºC

Denaturation 30 s 94 ºC
Annealing 1 min30 s 59 ºC 30
Extension 1 min 30 s 72º C

Final extension 10 min 72º C

Table No. 4 : The PCR cycling conditions to amplify blaTEM gene fragments.

Step
Program
blaSHV Cycles
Time Temperature

Initial denaturation 15 min 95 ºC

Denaturation 30 s 94 ºC
Annealing 1min 30 s 52 ºC 30
Extension 1 min 30 s 72º C

Final extension 1 min 30 s 72º C

Table No. 5 : The PCR cycling conditions to amplify blaSHV gene fragments

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LACTAMASE (ESBL) PRODUCING E.COLI ISOLATES WITH SPECIAL REFERENCE TO blaTEM ,
blaCTX AND blaSHV GENE FROM PATIENTS OF URINARY TRACT.
SEEJPH Volume XXVI, S1, 2025, ISSN: 2197-5248; Posted:05-01-2025

Step
Program
blaCTX Cycles
Time Temperature

Initial denaturation 15 min 95 ºC

Denaturation 30 s 94 ºC
Annealing 1min 30 s 52 ºC 30
Extension 1 min 30 s 72º C

Final extension 1 min 30 s 72º C

Table No. 6 : The PCR cycling conditions to amplify blaCTX gene fragments

For acquired blaSHV genes, the initial denaturation was at 95 °C for 15 min, 30 cycles of 94 °C
for 30 s, 52 °C for 1 min 30 s and 72 °C for 1 min 30 s, followed by extension of 72 °C for 1 min
30 s. The cyclic conditions for blaCTX gene, initial denaturation at 95 °C for 15 min, 30 cycles
of 94 °C for 30 s, 59 °C for 1 min 30 s and 72 °C for 1 min 30 s were followed by extension of
72 °C for 10 min.

The Agarose gel preparation and visualized by Gel Doc™ EZ Gel Documentation System
 The Agarose Gel Electrophoresis was performed in order to identify the Purified PCR
Product which was previously identified by its amplified DNA fragments.
 The resulting PCR product was subjected to 1 % agarose gel electrophoresis and visualized
by Gel Doc™ EZ Gel Documentation System (Bio-Rad Laboratories Inc., Hercules, CA,
USA).
 A 1 kb DNA Ladder (Thermo Fisher Scientific ™, Waltham, MA, USA) was used as the
marker to evaluate the PCR product of the sample.

STATISTIC ANALYSIS
Data along with statistic was recorded by the Microsoft Excel. The values were represented in
Numbers percentage and bar diagram..

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LACTAMASE (ESBL) PRODUCING E.COLI ISOLATES WITH SPECIAL REFERENCE TO blaTEM ,
blaCTX AND blaSHV GENE FROM PATIENTS OF URINARY TRACT.
SEEJPH Volume XXVI, S1, 2025, ISSN: 2197-5248; Posted:05-01-2025

RESULTS
In the present study out of the 1030 isolates there were 366 (35.5%) which showed the growth of
E. coli. In which 82 (22.5%) were phenotypically identified as ESBL producers and 284 (77.6%)
were Non-ESBL. Out of the 366 isolates, 110 (30%) were Males and 256 (69.9%) were Females
patients.
Table 7 shows that Out of 366 patients who were included in this study 110 (30%) were Male &
256 (69.9%) were Female patients.

Table 7: Distribution of Patients according to Gender (Male/ Female)

Gender No. of Isolates Percentage (%)

Male 110 30%

Female 256 69.9%

Total 366 100

Table 8 shows that the age group of 21–30 years old accounts for the greatest number of
instances (24.1%), while the age group of patients over 80 years old accounts for the fewest
(0.9%).

Table 8: Distribution of Patients according to Age group

61- 71- 81-

Age Group 0-10 11-20 21-30 31-40 41-50 51-60 70 80 90 Total

No. of
Isolates 37 40 88 54 50 56 23 14 3 366

Percentage 10.2% 10.7% 24.1% 15.1% 13.5% 15.3% 6.3% 3.9% 0.9% 100%

Table 9 shows that the overall susceptibility of ESBL isolates to various antibiotics was as
Follows: Ampicillin(17.20%), Ampicillin/Sulbactam (28.5%), Gentamycin (65.7%), Cefoxitin
(51.0%), Amikacin (80%), Ciprofloxacin (48%), Meropenem (97.2%), Ceftazidime(0%),
Ceftazidime/ clavunalate(100%), Piperacillin/tazobactam (85.7%), Ceftriaxone(0%),
Nitrofurantoin(100%), Tigecycillin(97.2%) and fosfomycin(97.2%).

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LACTAMASE (ESBL) PRODUCING E.COLI ISOLATES WITH SPECIAL REFERENCE TO blaTEM ,
blaCTX AND blaSHV GENE FROM PATIENTS OF URINARY TRACT.
SEEJPH Volume XXVI, S1, 2025, ISSN: 2197-5248; Posted:05-01-2025

Table 9: Antimicrobial Sensitivity & Resistivity of ESBL producing E. coli

Antibiotics Sensitivity Resistivity

Ceftazidime/clavunalte 100% 0%

Nitrofurantoin 100% 0%

Fosfomycin 97.20% 2.80%

Meropenem 97.20% 2.80%

Tigecycillin 97.20% 2.80%

Piperacillin/Tazobactam 85.70% 14.30%

Amikacin 80% 20%

Gentamycin 65.70% 34.30%

Cefoxitin 51% 49%

Ciprofloxacin 48% 52%

Ampicillin/Sulbactum 28.50% 71.50%

Ampicillin/Sulbactum 17.20% 82.80%

Ceftazime 0% 100%

Ceftriaxone 0% 100%

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LACTAMASE (ESBL) PRODUCING E.COLI ISOLATES WITH SPECIAL REFERENCE TO blaTEM ,
blaCTX AND blaSHV GENE FROM PATIENTS OF URINARY TRACT.
SEEJPH Volume XXVI, S1, 2025, ISSN: 2197-5248; Posted:05-01-2025

Figure No 4.: The DNA Extraction of the test isolates

L1, L2,L3L4-L6

Figure No. 5: The blaSHV 795 bp 795bp

L1 corresponds to the DNA Ladder; L2 corresponds to the positive Control; L3 Corresponds

to the Negative Control to blaSHV gene; L4-L6 are the sample positive for blaSHV gene

blaCTX
DNA Ladder
L1-L9,L10,L11-16,L19,20

544bp

Figure No. 6 : The Amplified DNA with PCR for blaCTX gene of E.coli . Lane -1-9 are
positive for CTX gene; Lane 10 is the DNA Ladder; Lane 11-16 and 19, 20 are CTX gene
positive; Lane 17 is the Negative control for CTX gene; Lane 18 is the positive control for
CTX gene

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LACTAMASE (ESBL) PRODUCING E.COLI ISOLATES WITH SPECIAL REFERENCE TO blaTEM ,
blaCTX AND blaSHV GENE FROM PATIENTS OF URINARY TRACT.
SEEJPH Volume XXVI, S1, 2025, ISSN: 2197-5248; Posted:05-01-2025

Figure No. 7 : The blaTEM 861 bp

L corresponds to the DNA Ladder; L1 corresponds to the positive Control; L2 Corresponds
to the Negative Control to blaTEM gene; L3-L7 are the sample positive for blaTEM gene

Table 10: Distribution of different genes in ESBL-producing E.coli isolates

Gene No. of Gene Detected Percentage

blaCTX 24 29.2%

blaSHV 13 15.8%

blaTEM 42 51.2%

In the current study out of the total 82 isolates there were 42 (51.2%) observed positive for blaTEM
gene, 24 (29.2% ) observed positive for blaCTX and 13 (15.8%) observed positive for blaSHV gene
13`(15.8%) .

DISCUSSION
Over the past 20 years, gram-negative bacteria that produce ESBL in particular, E. coli—have
emerged as significant global pathogens in both community-acquired and hospital-acquired
illnesses. It is advised to use β-lactam medications, such as carbapenems and long spectrum
cephalosporins, to treat enterobacterial infections There were initially only a few bacterial
species, but they are now rapidly expanding, and the maturity of conflict to extended-spectrum
cephalosporins in Gram-negative bacteria has been a major cause for concern [17].

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LACTAMASE (ESBL) PRODUCING E.COLI ISOLATES WITH SPECIAL REFERENCE TO blaTEM ,
blaCTX AND blaSHV GENE FROM PATIENTS OF URINARY TRACT.
SEEJPH Volume XXVI, S1, 2025, ISSN: 2197-5248; Posted:05-01-2025

Characteristic of antimicrobial resistance are the differences between regions, hospitals and even
departments. E. coli strains are susceptible to commonly used antimicrobial agents in treatment
of UTIs. However, antibiotic resistance of uropathogenic E. coli in UTIs is increasing worldwide
[18].
This study shows that out of 366 patients 82 (22.4%) ESBL 284 (77.5%) & Non-ESBL. And this
study also observed that maximum number of patients belong to age group of 21–30 years old 88
(24.1%) followed by 31-40 years. It was also observed that Female were more affected 256
(69.9%) than Male 110 (30%) . This study was parallel to the study performed by the other
research investigator where UTIs caused by ESBL-producing E. coli were overall far more
common among females [19,20].
In the present study it was observed that out of the total 82 isolates there were 42 (51.2%)
observed positive for blaTEM gene, 24 (29.2% ) observed positive for blaCTX and 13 (15.8%)
observed positive for blaSHV gene. This study was in accordance to the study performed by the
other research investigator M.C. El bouamri et al., where the ESBL production patterns
observed included single production of CTX-M (70%), SHV (12%) but in constrast with TEM
(0%) [21]. There was another study which was in support to the current study where the bla
CTX-M (77.4%; n = 377), bla TEM (54.4%; n = 265) and Ib-cr (52%; n = 253) genes and a low
proportion of bla SHV and qnr genes were observed [22].
There was another study by Sheetal verma et al., in 2022 where phenotypically positive ESBL
isolates, blaTEM (49.4%) was the most common genotype followed by blaCTX- M1 (31.97%),
blaOXA-1 (30.1%), and blaSHV(11.9%) either alone or in combination. This study was in
support to the present study [23]. In a study from Assam, CTX-M, TEM, and SHV were detected
in 54.4, 33.9, and 15.4% isolates, respectively [24]. There was another study was in alignment to
the current study where the blaCTX−M−1 (60.7%) was the most common among [25]. There
was another study which was in accordance to the current study where out of the collected strains
of ESBL-producing E. coli, had 81% blaTEM, 16.2% blaSHV, and 32.4% blaCTX-M genes.
Similarly, 64.7% blaTEM, 35.2% blaSHV, and 41.1% blaCTX-M genes existed in the isolates of
K. pneumoniae [26].
There were few studies from Central India which were also in accordance to the current study
and reported blaTEM gene most predominant followed by blaCTX-M and blaSHV [27].
More research examining the risk factors, diagnostic importance, and possible treatments for
acquired in the community illnesses triggered by these microbes is required because there are
relatively only a few alternatives for treating these infections . However, these patients usually
delay receiving the proper therapy, which could have negative clinical results. The fact is that we
have a good arsenal at our disposal to treat these infections, despite recent disagreements over if
a carbapenem antibiotic has to be employed for treating severe infections caused by ESBLs or
while certain β-lactamases/ β-lactamases inhibitor combinations remain suitable. It continues to
remain crucial for us to keep a close eye on ESBLs in patients isolates as well as in monitoring
because they have become widespread in healthcare isolates of Enterobacterales.
Common ESBL genes coding for isolates of K. pneumoniae and E. coli were determined as
CTX-M (cefotaximase that preferentially hydrolyzes cefotaxime), TEM (found and isolated in
the early 80s from Teminora who was a Greek patient), and SHV (for variable of sulphydryl
which was first observed in a single Klebsiella ozaenae strain retrieved in Germany) [28-30] .
These genes which are mediated by transposons, plasmids, or chromosomes are all sporadically
described all over the world.

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 TO STUDY THE MOLECULAR PROFILING AND ITS ASSOCIATION TO EXTENDED SPECTRUM Β-
LACTAMASE (ESBL) PRODUCING E.COLI ISOLATES WITH SPECIAL REFERENCE TO blaTEM ,
blaCTX AND blaSHV GENE FROM PATIENTS OF URINARY TRACT.
SEEJPH Volume XXVI, S1, 2025, ISSN: 2197-5248; Posted:05-01-2025

CONCLUSION
High antibiotic drug resistance makes it difficult to treat ESBL infections and pushes doctors to
use colistin and carbapenems more often. OXA-1, VEB, and PER-2 type β-lactamase are
emerging, while TEM, CTX-M-1, CTX-M-15, and SHV-type ESBL are rather common. The
proliferation of ESBL makes more careful monitoring necessary to put appropriate control
mechanisms in place.
Hospitals urgently need to adopt infection control bundles and checklists. Local antibiograms are
essential for selecting the most effective antibiotics for empirical treatments.

Declarations:
Conflicts of interest: There is not any conflict of interest associated with this study
Consent to participate: There is consent to participate.
Consent for publication: There is consent for the publication of this paper.
Authors' contributions: Author equally contributed the work.

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