Rapid and Accurate Detection of <i>Escherichia coli</i> and <i>Klebsiella pneumoniae</i> Strains Susceptible/Resistant to Cotrimoxazole through Evaluation of Cell Elongation
Trimethoprim-sulfamethoxazole is a well-known antibiotic that inhibits folic acid synthesis, a topic of renewed interest. Since resistant strains are increasingly more common, an early and accurate discrimination of susceptibility may assure confident therapy. Two morphological assays were performed...
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doaj-6c6c311a1a0d421ebac43b359db5f3c92021-07-01T00:15:38ZengMDPI AGAntibiotics2079-63822021-06-011072072010.3390/antibiotics10060720Rapid and Accurate Detection of <i>Escherichia coli</i> and <i>Klebsiella pneumoniae</i> Strains Susceptible/Resistant to Cotrimoxazole through Evaluation of Cell ElongationIsidoro López0Fátima Otero1Rebeca Guillén2María del Carmen Fernández3Germán Bou4Jaime Gosálvez5José Luis Fernández6Genetics Unit, INIBIC-Complejo Hospitalario Universitario A Coruña (CHUAC), 15006 A Coruña, SpainGenetics Unit, INIBIC-Complejo Hospitalario Universitario A Coruña (CHUAC), 15006 A Coruña, SpainGenetics Unit, INIBIC-Complejo Hospitalario Universitario A Coruña (CHUAC), 15006 A Coruña, SpainMicrobiology Division, INIBIC-Complejo Hospitalario Universitario A Coruña (CHUAC), 15006 A Coruña, SpainMicrobiology Division, INIBIC-Complejo Hospitalario Universitario A Coruña (CHUAC), 15006 A Coruña, SpainGenetics Unit, Facultad de Biología, Universidad Autónoma de Madrid, 28049 Madrid, SpainGenetics Unit, INIBIC-Complejo Hospitalario Universitario A Coruña (CHUAC), 15006 A Coruña, SpainTrimethoprim-sulfamethoxazole is a well-known antibiotic that inhibits folic acid synthesis, a topic of renewed interest. Since resistant strains are increasingly more common, an early and accurate discrimination of susceptibility may assure confident therapy. Two morphological assays were performed in <i>Escherichia coli</i> (<i>n</i> = 50; 27 non-susceptible) and <i>Klebsiella pneumoniae</i> (<i>n</i> = 52; 18 non-susceptible). First, the strains were incubated with the CLSI breakpoint of cotrimoxazole for 150 min, which induced cell lengthening in the susceptible strains. Second, the bacteria were incubated with mitomycin C (MMC) (0.5 mg/L) for 120 min to induce a SOS-linked cell enlargement higher than that obtained by cotrimoxazole alone. When cotrimoxazole was added 30 min before MMC, the inhibition of folic acid synthesis in the susceptible strain resulted in the suppression of MMC-induced extra elongation. In the non-susceptible strains, folic acid synthesis continued despite the antibiotic, so that the MMC-induced extra cell lengthening could not be impeded. Whereas the first assay resulted in five false negatives and four false positives of resistance, the results of the second assay matched those of the conventional antibiogram. This simple morphological procedure is performed in 2 h and 45 min and may allow a rapid selection of useful and relatively inexpensive therapy, thereby preserving the newer broad-spectrum antibiotics.https://www.mdpi.com/2079-6382/10/6/720cotrimoxazoletrimethoprim-sulfamethoxazoleantibiotic resistancecell lengthrapid assayGram-negative |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Isidoro López Fátima Otero Rebeca Guillén María del Carmen Fernández Germán Bou Jaime Gosálvez José Luis Fernández |
spellingShingle |
Isidoro López Fátima Otero Rebeca Guillén María del Carmen Fernández Germán Bou Jaime Gosálvez José Luis Fernández Rapid and Accurate Detection of <i>Escherichia coli</i> and <i>Klebsiella pneumoniae</i> Strains Susceptible/Resistant to Cotrimoxazole through Evaluation of Cell Elongation Antibiotics cotrimoxazole trimethoprim-sulfamethoxazole antibiotic resistance cell length rapid assay Gram-negative |
author_facet |
Isidoro López Fátima Otero Rebeca Guillén María del Carmen Fernández Germán Bou Jaime Gosálvez José Luis Fernández |
author_sort |
Isidoro López |
title |
Rapid and Accurate Detection of <i>Escherichia coli</i> and <i>Klebsiella pneumoniae</i> Strains Susceptible/Resistant to Cotrimoxazole through Evaluation of Cell Elongation |
title_short |
Rapid and Accurate Detection of <i>Escherichia coli</i> and <i>Klebsiella pneumoniae</i> Strains Susceptible/Resistant to Cotrimoxazole through Evaluation of Cell Elongation |
title_full |
Rapid and Accurate Detection of <i>Escherichia coli</i> and <i>Klebsiella pneumoniae</i> Strains Susceptible/Resistant to Cotrimoxazole through Evaluation of Cell Elongation |
title_fullStr |
Rapid and Accurate Detection of <i>Escherichia coli</i> and <i>Klebsiella pneumoniae</i> Strains Susceptible/Resistant to Cotrimoxazole through Evaluation of Cell Elongation |
title_full_unstemmed |
Rapid and Accurate Detection of <i>Escherichia coli</i> and <i>Klebsiella pneumoniae</i> Strains Susceptible/Resistant to Cotrimoxazole through Evaluation of Cell Elongation |
title_sort |
rapid and accurate detection of <i>escherichia coli</i> and <i>klebsiella pneumoniae</i> strains susceptible/resistant to cotrimoxazole through evaluation of cell elongation |
publisher |
MDPI AG |
series |
Antibiotics |
issn |
2079-6382 |
publishDate |
2021-06-01 |
description |
Trimethoprim-sulfamethoxazole is a well-known antibiotic that inhibits folic acid synthesis, a topic of renewed interest. Since resistant strains are increasingly more common, an early and accurate discrimination of susceptibility may assure confident therapy. Two morphological assays were performed in <i>Escherichia coli</i> (<i>n</i> = 50; 27 non-susceptible) and <i>Klebsiella pneumoniae</i> (<i>n</i> = 52; 18 non-susceptible). First, the strains were incubated with the CLSI breakpoint of cotrimoxazole for 150 min, which induced cell lengthening in the susceptible strains. Second, the bacteria were incubated with mitomycin C (MMC) (0.5 mg/L) for 120 min to induce a SOS-linked cell enlargement higher than that obtained by cotrimoxazole alone. When cotrimoxazole was added 30 min before MMC, the inhibition of folic acid synthesis in the susceptible strain resulted in the suppression of MMC-induced extra elongation. In the non-susceptible strains, folic acid synthesis continued despite the antibiotic, so that the MMC-induced extra cell lengthening could not be impeded. Whereas the first assay resulted in five false negatives and four false positives of resistance, the results of the second assay matched those of the conventional antibiogram. This simple morphological procedure is performed in 2 h and 45 min and may allow a rapid selection of useful and relatively inexpensive therapy, thereby preserving the newer broad-spectrum antibiotics. |
topic |
cotrimoxazole trimethoprim-sulfamethoxazole antibiotic resistance cell length rapid assay Gram-negative |
url |
https://www.mdpi.com/2079-6382/10/6/720 |
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