Molecular characterization of extended-spectrum β-lactamases in clinical Escherichia coli and Klebsiella pneumoniae isolates from Surabaya, Indonesia

Severin, Juliëtte A.; Mertaniasih, Ni Made; Kuntaman, Kuntaman; Lestari, Endang Sri; Purwanta, Marijam; Toom, Nicole Lemmens-den; Duerink, D. Offra; Hadi, Usman; Belkum, Alex van; Verbrugh, Henri A.; Goessens, W. H. F.

doi:10.1093/jac/dkp471

Cited by 53 publications

(43 citation statements)

References 8 publications

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“…PCRs were used as previously described to amplify several ␤-lactamase genes coding for ESBLs (bla CTX-M , bla TEM , and bla SHV ), plasmid-encoded AmpCs (bla CMY , bla FOX , bla MOX , bla DHA , bla LAT , bla ACT , and bla MIR ), and oxacillinases (24)(25)(26)(27)(28)(29). Additionally, we tested for the carbapenemase genes bla VIM , bla IMP , and bla KPC (30,31).…”

Section: Patient and Isolatesmentioning

confidence: 99%

“…Additionally, we tested for the carbapenemase genes bla VIM , bla IMP , and bla KPC (30,31). Sequencing of PCR products was performed using a 3100 ABI Prism genetic analyzer (Applied Biosystems, Foster City, CA) with an additional primer for the CTX-M-1 group (28). For sequencing of the CMY-2 gene, primers described by Bauernfeind et al were used (32).…”

Section: Patient and Isolatesmentioning

confidence: 99%

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Antibiotic Trapping by Plasmid-Encoded CMY-2 β-Lactamase Combined with Reduced Outer Membrane Permeability as a Mechanism of Carbapenem Resistance in Escherichia coli

Goessens

Bij

Boxtel

et al. 2013

Antimicrob Agents Chemother

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A liver transplant patient was admitted with cholangitis, for which meropenem therapy was started. Initial cultures showed a carbapenem-susceptible (CS) Escherichia coli strain, but during admission, a carbapenem-resistant (CR) E. coli strain was isolated. Analysis of the outer membrane protein profiles showed that both CS and CR E. coli lacked the porins OmpF and OmpC. Furthermore, PCR and sequence analysis revealed that both CS and CR E. coli possessed bla CTX-M-15 and bla OXA-1 . The CR E. coli strain additionally harbored bla CMY-2 and demonstrated a >15-fold increase in ␤-lactamase activity against nitrocefin, but no hydrolysis of meropenem was detected. However, nitrocefin hydrolysis appeared strongly inhibited by meropenem. Furthermore, the CMY-2 enzyme demonstrated lower electrophoretic mobility after its incubation either in vitro or in vivo with meropenem, indicative of its covalent modification with meropenem. The presence of the acyl-enzyme complex was confirmed by mass spectrometry. By transformation of the CMY-2-encoding plasmid into various E. coli strains, it was established that both porin deficiency and high-level expression of the enzyme were needed to confer meropenem resistance. In conclusion, carbapenem resistance emerged by a combination of elevated ␤-lactamase production and lack of porin expression. Due to the reduced outer membrane permeability, only small amounts of meropenem can enter the periplasm, where they are trapped but not degraded by the large amount of the ␤-lactamase. This study, therefore, provides evidence that the mechanism of "trapping" by CMY-2 ␤-lactamase plays a role in carbapenem resistance.

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Section: Patient and Isolatesmentioning

confidence: 99%

Section: Patient and Isolatesmentioning

confidence: 99%

Antibiotic Trapping by Plasmid-Encoded CMY-2 β-Lactamase Combined with Reduced Outer Membrane Permeability as a Mechanism of Carbapenem Resistance in Escherichia coli

Goessens

Bij

Boxtel

et al. 2013

Antimicrob Agents Chemother

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“…CTX-M types have been isolated and these have been divided into 5 clusters on the basis of amino-acid sequences: CTX-M-1, CTX-M-2, CTX-M-8, CTX-M-9, and CTX-M-25 [5]. In many countries CTX-M blactamases are the most prevalent type of ESBL, such as in Switzerland (5), Turkey (6), and Indonesia (7).…”

Section: Introductionmentioning

confidence: 99%

High prevalence of CTX-M-15 and first report of CTX-M-3, CTX-M-22, CTX-M-28 and plasmid-mediated AmpC beta-lactamase producing Enterobacteriaceae causing urinary tract infections in Bosnia and Herzegovina in hospital and community settings

Ibrahimagić

Bedenić

Kamberović

et al. 2015

Journal of Infection and Chemotherapy

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“…5 Di Asia pada tahun 1999 diperkirakan bahwa sekitar 12-24% isolat Eschericia coli positif ESBL, sementara untuk Klebsiella pneumoniae penghasil ESBL jumlahnya kurang dari 5%. 6 Persentase serupa ditemukan pada penelitian di RSP Dr. Soetomo Surabaya pada tahun 2010, 7 sementara penelitian di Bandung pada tahun 2012 didapatkan angka yang sangat tinggi, yaitu 72% isolat Klebsiella pneumoniae sampel pasien yang menghasilkan ESBL. 8 Penggunaan antibiotik yang sering di masyarakat pun dapat turut meningkatkan risiko penyebaran ESBL.…”

Section: Pendahuluanunclassified

“…Extended-spectrum beta-lactamase dihasilkan sebagian besar oleh bakteri gram negatif dari famili Enterobacteriaceae, terutama pada spesies K. pneumoniae dan E. coli. 7 Enterobacteriaceae merupakan salah satu bakteri yang paling sering mengakibatkan penyakit pada manusia, seperti infeksi pada sistem saraf pusat, saluran napas bawah, aliran darah, saluran kemih, dan saluran pencernaan. Bakteri pada famili ini mempunyai kemampuan untuk menyebar dengan mudah di antara manusia dan dapat memperoleh materi genetik melalui transfer gen horizontal yang kebanyakan dimediasi oleh plasmid.…”

Section: Pendahuluanunclassified

Loop-Mediated Isothermal Amplification untuk Mendeteksi Gen blaTEM Sebagai Penyandi Extended-Spectrum Beta-Lactamase pada Isolat Enterobacteriaceae

Wilopo

Sudigdoadi²,

Sahiratmadja³

et al. 2015

mkb

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AbstrakExtended-spectrum beta-lactamase (ESBL) adalah suatu jenis enzim beta-laktamase yang mampu menghidrolisis penisilin, sefalosprin, dan monobaktam yang dapat dihambat oleh asam klavulanat. Enzim ini disandi oleh banyak gen, salah satunya adalah bla TEM . Untuk mengamplifikasi gen bla TEM selain digunakan metode polymerase chain reaction (PCR) dapat pula dilakukan metode loop-mediated isothermal amplification (LAMP) yang membutuhkan peralatan lebih sederhana dengan prosedur yang cepat dan pembacaan hasil yang lebih mudah. Penelitian ini merupakan uji diagnostik yang bertujuan menilai sensitivitas dan spesifisitas metode LAMP serta melihat kesesuaian hasil antara metode LAMP dan metode PCR dalam mendeteksi gen bla TEM Loop-Mediated Isothermal Amplification for Detecting bla TEM Gene that Encodes Extended-Spectrum Beta-Lactamase in Enterobacteriaceae IsolatesAbstract Extended-spectrum beta-lactamase (ESBL) is a beta-lactamase enzyme that is capable of hydrolyzing penicillin, cephalosporin, and monobactam, and can be inhibited by clavulanic acid. This enzyme is encoded by multiple genes, one of them is blaTEM. Polymerase chain reaction (PCR) is one of the DNA amplification methods that are frequently used; however, there are other methods that can be used including, among others, loop-mediated isothermal amplification (LAMP). LAMP requires simple equipment with quicker and easy-to-read results compared to PCR. This study was a diagnostic test to explore the sensitivity and specificity of LAMP method compared to PCR in detecting blaTEM gene. Furthermore, the concordance between LAMP and PCR methods was assessed. A total of 92 Enterobacteriaceae isolates were examined by PCR and LAMP methods and compared. The result showed that the LAMP method had a sensitivity of 91.4% and a specificity of 91.2% with a concordance value (kappa) of 85.4%. In conclusion, LAMP method has a good validity and a very good conformity compared to the PCR method. Therefore, LAMP method can be used as an alternative diagnostic test, especially in limited settings. [MKB. 2015;47(4):242-9]

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Molecular characterization of extended-spectrum β-lactamases in clinical Escherichia coli and Klebsiella pneumoniae isolates from Surabaya, Indonesia

Abstract: Indonesia is another developing country affected by the emergence and spread of bacterial strains harbouring ESBL genes, including the CTX-M-15-producing B2-E. coli O25b-ST131 clone.

Cited by 53 publications

References 8 publications

Antibiotic Trapping by Plasmid-Encoded CMY-2 β-Lactamase Combined with Reduced Outer Membrane Permeability as a Mechanism of Carbapenem Resistance in Escherichia coli

Antibiotic Trapping by Plasmid-Encoded CMY-2 β-Lactamase Combined with Reduced Outer Membrane Permeability as a Mechanism of Carbapenem Resistance in Escherichia coli

High prevalence of CTX-M-15 and first report of CTX-M-3, CTX-M-22, CTX-M-28 and plasmid-mediated AmpC beta-lactamase producing Enterobacteriaceae causing urinary tract infections in Bosnia and Herzegovina in hospital and community settings

Loop-Mediated Isothermal Amplification untuk Mendeteksi Gen blaTEM Sebagai Penyandi Extended-Spectrum Beta-Lactamase pada Isolat Enterobacteriaceae

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