Comparing MALDI-ToFMass Spectrometry with Molecular and Biochemical Methods in Identifying Enterococcus Faecium and Enterococcus FaecalisIsolated from Clinical Samples
Abstract Background and Objective: Enterococci are Gram-positive members of human gastrointestinal flora,in Dairy products and environment. they have emerged as important causes of opportunistic nosocomial infections in recent years. In this study we aimed to investigat and compare the efficiency of...
| Main Authors: | , , |
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| Format: | Article |
| Language: | English |
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Golestan University of Medical Sciences
2013-04-01
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| Series: | Medical Laboratory Journal |
| Subjects: | |
| Online Access: | http://mlj.goums.ac.ir/article-1-251-en.html |
| _version_ | 1823943421964845056 |
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| author | h SamadiKafil m MohebatiMobarez m ForouzandehMoghadam |
| author_facet | h SamadiKafil m MohebatiMobarez m ForouzandehMoghadam |
| author_sort | h SamadiKafil |
| collection | DOAJ |
| description | Abstract
Background and Objective: Enterococci are Gram-positive members of human gastrointestinal flora,in Dairy products and environment. they have emerged as important causes of opportunistic nosocomial infections in recent years. In this study we aimed to investigat and compare the efficiency of MALDI-TOFmass spectroscopy method through Biochemical and Molecular methods for detecting Enterococcus faecalis and Enterococcus faecium.
Materials and Methods:seventhy five clinical samples were collected for biochemical, molecular and mass spectroscopy investigations. Samples were treated with Esculin hydrolysis, Catalase, Pyrrolidonylaminopeptidase, 6.5% NaCl solution, motility, 0.04% Tellurite, L-Arabinose and Sorbitol. Using specific primesallele specific PCR was used.The samples were then analyzed by MALDI-TOF mass spectroscopy and Biotyper 3 software.
Results:Enterococcus faecium andEnterococcus faecaliswere detected in thirty and forty two samples, respectively whereas three samples showed both bacterial infections. Using biochemical analysis, two E.faecium isolates were Arabinose negative and one E. faecalis isolates was Telliurite negative. All sampleswere showed correct bands in PCR results but twoof them didn't show clear bands(on agarose gel). In mass spectroscopy analysis all strains were correctly detected and well defined.
Conclusion: According to our results, MALDI-TOF mass spectrometry in comparison with Molecular and Biochemical Methodscould be a reliable and accurate method that can easily and quickly identify and differentiate Enterococcus faecium and Enterococcus faecalisin clinical samples.
Key words:Enterococcus faecalis, Enterococcus faecium, MALDI-TOFmass spectrometry,PCR |
| first_indexed | 2024-12-17T07:58:45Z |
| format | Article |
| id | doaj.art-be5a3755949a45cb922ea02ede2cf889 |
| institution | Directory Open Access Journal |
| issn | 2538-4449 |
| language | English |
| last_indexed | 2024-12-17T07:58:45Z |
| publishDate | 2013-04-01 |
| publisher | Golestan University of Medical Sciences |
| record_format | Article |
| series | Medical Laboratory Journal |
| spelling | doaj.art-be5a3755949a45cb922ea02ede2cf8892022-12-21T21:57:37ZengGolestan University of Medical SciencesMedical Laboratory Journal2538-44492013-04-017116Comparing MALDI-ToFMass Spectrometry with Molecular and Biochemical Methods in Identifying Enterococcus Faecium and Enterococcus FaecalisIsolated from Clinical Samplesh SamadiKafil0m MohebatiMobarez1m ForouzandehMoghadam2 ,Department of Medical Bacteriology, Faculty of Medical Sciences, Tarbiatmodares University Department of Medical Bacteriology, Faculty of Medical Sciences, Tarbiatmodares University Department of Biotechnology, Faculty of Medical Sciences, TarbiatModares University, Tehran, Iran Abstract Background and Objective: Enterococci are Gram-positive members of human gastrointestinal flora,in Dairy products and environment. they have emerged as important causes of opportunistic nosocomial infections in recent years. In this study we aimed to investigat and compare the efficiency of MALDI-TOFmass spectroscopy method through Biochemical and Molecular methods for detecting Enterococcus faecalis and Enterococcus faecium. Materials and Methods:seventhy five clinical samples were collected for biochemical, molecular and mass spectroscopy investigations. Samples were treated with Esculin hydrolysis, Catalase, Pyrrolidonylaminopeptidase, 6.5% NaCl solution, motility, 0.04% Tellurite, L-Arabinose and Sorbitol. Using specific primesallele specific PCR was used.The samples were then analyzed by MALDI-TOF mass spectroscopy and Biotyper 3 software. Results:Enterococcus faecium andEnterococcus faecaliswere detected in thirty and forty two samples, respectively whereas three samples showed both bacterial infections. Using biochemical analysis, two E.faecium isolates were Arabinose negative and one E. faecalis isolates was Telliurite negative. All sampleswere showed correct bands in PCR results but twoof them didn't show clear bands(on agarose gel). In mass spectroscopy analysis all strains were correctly detected and well defined. Conclusion: According to our results, MALDI-TOF mass spectrometry in comparison with Molecular and Biochemical Methodscould be a reliable and accurate method that can easily and quickly identify and differentiate Enterococcus faecium and Enterococcus faecalisin clinical samples. Key words:Enterococcus faecalis, Enterococcus faecium, MALDI-TOFmass spectrometry,PCRhttp://mlj.goums.ac.ir/article-1-251-en.htmlenterococcus faecalisenterococcus faeciummaldi-tofmass spectrometrypcr |
| spellingShingle | h SamadiKafil m MohebatiMobarez m ForouzandehMoghadam Comparing MALDI-ToFMass Spectrometry with Molecular and Biochemical Methods in Identifying Enterococcus Faecium and Enterococcus FaecalisIsolated from Clinical Samples Medical Laboratory Journal enterococcus faecalis enterococcus faecium maldi-tofmass spectrometry pcr |
| title | Comparing MALDI-ToFMass Spectrometry with Molecular and Biochemical Methods in Identifying Enterococcus Faecium and Enterococcus FaecalisIsolated from Clinical Samples |
| title_full | Comparing MALDI-ToFMass Spectrometry with Molecular and Biochemical Methods in Identifying Enterococcus Faecium and Enterococcus FaecalisIsolated from Clinical Samples |
| title_fullStr | Comparing MALDI-ToFMass Spectrometry with Molecular and Biochemical Methods in Identifying Enterococcus Faecium and Enterococcus FaecalisIsolated from Clinical Samples |
| title_full_unstemmed | Comparing MALDI-ToFMass Spectrometry with Molecular and Biochemical Methods in Identifying Enterococcus Faecium and Enterococcus FaecalisIsolated from Clinical Samples |
| title_short | Comparing MALDI-ToFMass Spectrometry with Molecular and Biochemical Methods in Identifying Enterococcus Faecium and Enterococcus FaecalisIsolated from Clinical Samples |
| title_sort | comparing maldi tofmass spectrometry with molecular and biochemical methods in identifying enterococcus faecium and enterococcus faecalisisolated from clinical samples |
| topic | enterococcus faecalis enterococcus faecium maldi-tofmass spectrometry pcr |
| url | http://mlj.goums.ac.ir/article-1-251-en.html |
| work_keys_str_mv | AT hsamadikafil comparingmalditofmassspectrometrywithmolecularandbiochemicalmethodsinidentifyingenterococcusfaeciumandenterococcusfaecalisisolatedfromclinicalsamples AT mmohebatimobarez comparingmalditofmassspectrometrywithmolecularandbiochemicalmethodsinidentifyingenterococcusfaeciumandenterococcusfaecalisisolatedfromclinicalsamples AT mforouzandehmoghadam comparingmalditofmassspectrometrywithmolecularandbiochemicalmethodsinidentifyingenterococcusfaeciumandenterococcusfaecalisisolatedfromclinicalsamples |