Journal of Medicinal and Chemical Sciences

By Subject

Current Issue

By Issue

By Author

Author Index

Keyword Index

About Journal

Aims and Scope

Publication Ethics

Indexing and Abstracting

Peer Review Process

Related Links

Reviewers

FAQ

News

Molecular Detection of bla TEM and bla CTX-M Genes in Clinical and Food-Borne Klebsiella pneumoniae Isolates

Document Type : Original Article

Authors

Department of Biology, College of Science, University of Baghdad, Baghdad, Iraq

Abstract

To isolate Klebsiella pneumoniae, two hundred clinical and food samples were cultivated on differential and selective culture media. 120 bacterial isolates were isolated from the food and clinical samples. On MacConkey agar, 81 isolates of lactose fermenters (67.5%) and 39 isolates of non-lactose fermenters (32.5%) were found. Out of 120 bacterial isolates discovered in clinical and food sources, 32 were K. pneumoniae isolates, including 14 from food sources and 18 from clinical sources. K. pneumoniae was determined to be the species of bacteria isolated based on biochemical testing, morphological characteristics, and cultural identities. This conclusion was confirmed by the Vitek system. The findings of this investigation, which used phenotypic methods, revealed a high frequency of isolates of K. pneumoniae that produce Extended Spectrum B-Lactamase (ESBL) from patients in Iraq and a low frequency of isolates from food sources. After bacterial DNA was isolated, pure DNA samples with concentrations between 76 and 91 ng/ml were produced. Utilizing particular primers for the genes bla TEM and bla CTX-M, this work carried out the molecular detection of two ESBL-coding genes. According to the findings of this study, bla TEM was only found in clinical isolates at a detection rate of 27.77% and bla CTX-M at 38.88%. Isolates from food sources tested negative for bla TEM and bla CTX-M.

Graphical Abstract

Molecular Detection of bla TEM and bla CTX-M Genes in Clinical and Food-Borne Klebsiella pneumoniae Isolates

Keywords

Full Text

Introduction

Due to the misuse of antibiotics, the incidence of multidrug-resistant (MDR) human pathogenic microorganisms and their genetic determinants has grown dramatically in clinical and environmental contexts. As a result, infection treatment choices become restricted, especially when MDR bacteria possess genes encoding extended range antibiotic resistance. One of the most important resistance mechanisms among Enterobacteriaceae is the production of β-lactamases, including extended-spectrum- and AmpC-lactamases [1]. According to an epidemiological research, Enterobacteriaceae that produce extended-spectrum-lactamase (ESBL), particularly Escherichia coli and Klebsiella pneumoniae, are the leading sources of community-acquired illnesses [2]. CTX-M, TEM, and SHV genes are the most common and clinically relevant ESBL genes, with CTX-M enzymes emerging as the most common. CTX-M is classified into five groups based on their amino acid identities: CTX-M-1, CTX-M-2, CTX-M-8, CTX-M-9, and CTX-M-25. K. pneumoniae typically generates all three kinds of enzymes, but the advent of carbapenemase-producing K. pneumoniae and colistin-resistant K. pneumoniae has caused the most recent public health worry [3]. Extended-spectrum beta lactamases (ESBLs) have played a major role in the clinical area in recent decades in the wide-ranging and complicated world of B-lactamases. Their significant stems the fact that they greatly broaden the range of the earlier B-lactamases, allowing them to hydrolyze all penicillins, cephalosporins (excluding cephamycins) and aztreonam. Furthermore, most ESBL genes are located on plasmids that confer antibiotic resistance to many types of antibiotics and are easily transferred between and among bacterial pathogens [4]. Extended spectrum lactamase, is a Gram-negative bacterium enzyme imparting resistance to penicillins, cephalosporins, and monobactams, provides a unique therapeutic problem. In Enterobacteriaceae from retail food, extended range TEM-, SHV-, and CTX-M type enzymes have been found [5].

Materials and Methods

Bacterial isolation

The collected clinical

References
[1]. Richter L., Du Plessis E.M., Duvenage S., Korsten L., Occurrence, phenotypic and molecular characterization of extended-spectrum-and AmpC-β-Lactamase producing Enterobacteriaceae isolated from selected commercial spinach supply chains in South Africa, Frontiers in microbiology, 2020, 11:638 [Crossref], [Google Scholar], [Publisher]
[2]. Pillai D., Kalasseril S.G., Genetic diversity and prevalence of extended spectrum beta-lactamase producing Escherichia coli and Klebsiella pneumoniae in aquatic environment receiving untreated hospital effluents, 2022, 1 [Crossref], [Google Scholar], [Publisher]
[3]. Founou L.L., Founou R.C., Allam M., Ismail A., Djoko C.F., Essack S.Y., Genome sequencing of extended-spectrum β-lactamase (ESBL)-producing Klebsiella pneumoniae isolated from pigs and abattoir workers in Cameroon, Frontiers in microbiology, 2018, 9:188 [Crossref], [Google Scholar], [Publisher]
[4]. Bora A., Hazarika N.K., Shukla S.K., Prasad K.N., Sarma J.B., Ahmed G., Prevalence of blaTEM, blaSHV and blaCTX-M genes in clinical isolates of Escherichia coli and Klebsiella pneumoniae from Northeast India, Indian journal of Pathology and Microbiology, 2014, 57:249 [Google Scholar], [Publisher]
[5]. Dsani E., Afari E.A., Danso-Appiah A., Kenu E., Kaburi B.B., Egyir B., Antimicrobial resistance and molecular detection of extended spectrum β-lactamase producing Escherichia coli isolates from raw meat in Greater Accra region, Ghana, BMC microbiology, 2020, 20:253 [Crossref], [Google Scholar], [Publisher]
[6]. MacFaddin J.F., Biochemical tests for identification of medical bacteria, 3rd ed, Lipicott Williams and Wilkins USA, 2000 [Crossref], [Google Scholar], [Publisher]
[7]. Maniatis T., Fritsch E., Sambrook J., Molecular Cloning: a Laboratory Manual, Cold Spring Harbor Laboratory, New York, 1982 [Google Scholar]
[8]. Elazhary M.A., Saheb S.A., Roy R.S., Lagacé A., A simple procedure for the preliminary identification of aerobic gram negative intestinal bacteria with special reference to the Enterobacteriaceae, Canadian journal of comparative medicine, 1973, 37:43 [Google Scholar], [Publisher]
[9]. Wyres K.L., Lam M., Holt K.E., Population genomics of Klebsiella pneumonia, Nature Reviews Microbiology, 2020, 18:344 [Crossref], [Google Scholar], [Publisher]
[10]. Bagley S.T., Seidler R.J., Primary Klebsiella identification with MacConkey-inositol-carbenicillin agar, Applied and Environmental Microbiology, 1978, 36:536 [Crossref], [Google Scholar], [Publisher]
[11]. Humphries R., Bobenchik A.M., Hindler J.A., Schuetz A.N., Overview of Changes to the Clinical and Laboratory Standards Institute Performance Standards for Antimicrobial Susceptibility Testing, M100, 31st Edition. J Clin Microbiol., 2021, 59:e0021321 [Crossref], [Google Scholar], [Publisher]
[12]. Al-khazraji S.M., Hossain M.H., Hassoon A.S., Estimation of some Bioactive substances and Antibacterial activity of Zingiber officinale (Ginger) Extract. Journal of Biomedicine and Biochemistry, 2022, 1:29 [Google Scholar], [Publisher]
[13]. Castanheira M., Simner P.J., Bradford P.A., Extended-spectrum β-lactamases: An update on their characteristics, epidemiology and detection, JAC-antimicrobial resistance, Extended-spectrum β-lactamases: An update on their characteristics, epidemiology and detection, 2021, 3:dlab092 [Crossref], [Google Scholar], [Publisher]
[14]. Dadgostar P., Antimicrobial resistance: implications and costs. Infection and Drug Resistance, 2019, 12:3903 [Crossref], [Google Scholar], [Publisher]
[15]. Akanbi O.E., Njom H.A., Fri J., Otigbu A.C., Clarke A.M., Antimicrobial Susceptibility of Staphylococcus aureus Isolated from Recreational Waters and Beach Sand in Eastern Cape Province of South Africa. Int J Environ Res Public Health. 2017, 14:1001 [Crossref], [Google Scholar], [Publisher]
[16]. Gurung R.R., Maharjan P., Chhetri G.G., Antibiotic resistance pattern of Staphylococcus aureus with reference to MRSA isolates from pediatric patients. Future science OA, 2020, 6: FSO464 [Google Scholar], [Publisher]
[17]. Bush K., Past and present perspectives on β-lactamases. Antimicrobial agents and chemotherapy, 2018, 62:e01076-18 [Crossref], [Google Scholar], [Publisher]
[18]. Wilfinger W.W., Mackey K., Chomczynski P., Effect of pH and ionic strength on the spectrophotometric assessment of nucleic acid purity, Biotechniques, 1997, 22:474 [Crossref], [Google Scholar], [Publisher]
[19]. Liu C.M., Stegger M., Aziz M., Johnson T.J., Waits K., Nordstrom L., Gauld L., Weaver B., Rolland D., Statham S., Horwinski J., Escherichia coli ST131-H 22 as a Foodborne Uropathogen, MBio, 2018, 9:e00470 [Crossref], [Google Scholar], [Publisher]
[20]. Datta N., Kontomichalou P., Penicillinase synthesis controlled by infectious R factors in Enterobacteriaceae, Nature, 1965, 208:239 [Google Scholar]
[21]. Medeiros A.A., β-Lactamases, British Medical Bulletin, 1984, 40:18 [Crossref], [Google Scholar], [Publisher]
[22]. Maleki N., Tahanasab Z., Mobasherizadeh S., Rezaei A., Faghri J., Prevalence of CTX-M and TEM β-lactamases in Klebsiella pneumoniae isolates from patients with urinary tract infection, Al-Zahra hospital, Isfahan, Iran, Advanced biomedical research, 2018, 7:10 [Crossref], [Google Scholar]
[23]. Paterson D.L., Hujer K.M., Hujer A.M., Yeiser B., Bonomo M.D., Rice L.B., International Klebsiella Study Group, Extended-spectrum β-lactamases in Klebsiella pneumoniae bloodstream isolates from seven countries: dominance and widespread prevalence of SHV-and CTX-M-type β-lactamases, Antimicrobial agents and chemotherapy, 2003, 47:3554 [Crossref], [Google Scholar], [Publisher]
[24]. Ibrahim M.E., Algak T.B., Abbas M., Elamin B.K., Emergence of bla TEM, bla CTX-M, bla SHV and bla OXA genes in multidrug-resistant Enterobacteriaceae and Acinetobacter baumannii in Saudi Arabia, Experimental and Therapeutic Medicine, 2021, 22:1 [Crossref], [Google Scholar], [Publisher]
[25]. Ibrahim A.S., Youssef N., Prevalence of CTX-M, TEM and SHV beta-lactamases in clinical isolates of Escherichia coli and Klebsiella Pneumoniae isolated from Aleppo University Hospitals, Aleppo, Syria, Archives of Clinical Infectious Diseases, 2015, 10:e22540 [Crossref], [Google Scholar], [Publisher].‏
[26]. Su X., Yan X., Li Y., Zhang D., Li L., Geng Y., Liu S., Identification of extended-spectrum beta-lactamase (CTX-M)-producing Klebsiella pneumoniae belonging to ST37, ST290, and ST2640 in captive giant pandas, BMC Veterinary Research, 2022, 18:186 [Crossref], [Google Scholar], [Publisher]
[27]. Jasim S.A., Yasin G., Cartono C., Sevbitov A., Shichiyakh R.A., Al-Husseini Y., Mustafa Y.F., Jalil A.T., Iswanto A., Survey of ground beetles inhabiting agricultural crops in south-east Kazakhstan, Brazilian Journal of Biology, 2022, 84: [Crossref], [Google Scholar], [Publisher]
[28]. Salahdin O.D., Sayadi H., Solanki R., Parra R.M.R., Al-Thamir M., Jalil A.T., Izzat S.E., Hammid A.T., Arenas L.A.B., Kianfar E., Graphene and carbon structures and nanomaterials for energy storage, Applied Physics A, 2022, 128:703 [Crossref], [Google Scholar], [Publisher] 
[29]. Gunawan W., Rudiansyah M., Sultan M.Q., Ansari M.J., Izzat S.E., Al Jaber M.S., Kzar H.H., Mustafa Y.F., Hammid A.T., Jalil A.T., Aravindhan S., Effect of tomato consumption on inflammatory markers in health and disease status: A systematic review and meta-analysis of clinical trials. Clinical Nutrition ESPEN, 2022, 50:93 [Crossref], [Google Scholar], [Publisher]
[30]. Fitriyah A., Nikolenko D.A., Abdelbasset W.K., Maashi M.S., Jalil A.T., Yasin G., Abdulkadhm M.M., Samieva G.U., Lafta H.A., Abed A.M., Amaral L.S., Exposure to ambient air pollution and osteoarthritis; an animal study, Chemosphere, 2022, 301:134698 [Crossref], [Google Scholar], [Publisher]
[31]. Sivaraman R., Patra I., Opulencia M.J.C., Sagban R., Sharma H., Jalil A.T., Ebadi A.G., Evaluating the potential of graphene-like boron nitride as a promising cathode for Mg-ion batteries, Journal of Electroanalytical Chemistry, 2022, 917:116413 [Crossref], [Google Scholar], [Publisher]
Journal of Medicinal and Chemical Sciences
Volume 6, Issue 7
July 2023
Pages 1706-1713
Files
History
  • Receive Date: 30 November 1999
  • Accept Date: 30 November 1999
Share
How to cite
Statistics