Matrix-assisted laser desorption-ionization-time-of-flight mass spectrometry as a reliable proteomic method for characterization of Escherichia coli and Salmonella isolates
Aim: Identification of pathogenic clinical bacterial isolates is mainly dependent on phenotypic and genotypic characteristics of the microorganisms. These conventional methods are costive, time-consuming, and need special skills and training. An alternative, mass spectral (proteomics) analysis metho...
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| Format: | Article |
| Language: | English |
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Veterinary World
2017-09-01
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| Series: | Veterinary World |
| Subjects: | |
| Online Access: | http://www.veterinaryworld.org/Vol.10/September-2017/12.pdf |
| _version_ | 1818589518545027072 |
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| author | Waleed S. Shell Mahmoud Lotfy Sayed Fatma Mohamed Gad Allah Fatma Elzahraa Mohamed Gamal Afaf Ahmed Khder A. A. Samy Abde Hakam M. Ali |
| author_facet | Waleed S. Shell Mahmoud Lotfy Sayed Fatma Mohamed Gad Allah Fatma Elzahraa Mohamed Gamal Afaf Ahmed Khder A. A. Samy Abde Hakam M. Ali |
| author_sort | Waleed S. Shell |
| collection | DOAJ |
| description | Aim: Identification of pathogenic clinical bacterial isolates is mainly dependent on phenotypic and genotypic characteristics of the microorganisms. These conventional methods are costive, time-consuming, and need special skills and training. An alternative, mass spectral (proteomics) analysis method for identification of clinical bacterial isolates has been recognized as a rapid, reliable, and economical method for identification. This study was aimed to evaluate and compare the performance, sensitivity and reliability of traditional bacteriology, phenotypic methods and matrix-assisted laser desorption-ionization-time-of-flight mass spectrometry (MALDI-TOF MS) in the identification of clinical Escherichia coli and Salmonella isolates recovered from chickens.
Materials and Methods: A total of 110 samples (cloacal, liver, spleen, and/or gall bladder) were collected from apparently healthy and diseased chickens showing clinical signs as white chalky diarrhea, pasty vent, and decrease egg production as well as freshly dead chickens which showing postmortem lesions as enlarged liver with congestion and enlarged gall bladder from different poultry farms.
Results: Depending on colonial characteristics and morphological characteristics, E. coli and Salmonella isolates were recovered and detected in only 42 and 35 samples, respectively. Biochemical identification using API 20E identification system revealed that the suspected E. coli isolates were 33 out of 42 of colonial and morphological identified E. coli isolates where Salmonella isolates were represented by 26 out of 35 of colonial and morphological identified Salmonella isolates. Serological identification of isolates revealed that the most predominant E. coli serotypes were O1 and O78 while the most predominant Salmonella serotype of Salmonella was Salmonella Pullorum. All E. coli and Salmonella isolates were examined using MALDI-TOF MS. In agreement with traditional identification, MADI-TOF MS identified all clinical bacterial samples with valid scores as E. coli and Salmonella isolates except two E. coli isolates recovered from apparently healthy and diseased birds, respectively, with recovery rate of 93.9% and 2 Salmonella isolates recovered from apparently healthy and dead birds, respectively, with recovery rate of 92.3%.
Conclusion: Our study demonstrated that Bruker MALDI-TOF MS Biotyper is a reliable rapid and economic tool for the identification of Gram-negative bacteria especially E. coli and Salmonella which could be used as an alternative diagnostic tool for routine identification and differentiation of clinical isolates in the bacteriological laboratory. MALDI-TOF MS need more validation and verification and more study on the performance of direct colony and extraction methods to detect the most sensitive one and also need using more samples to detect sensitivity, reliability, and performance of this type of bacterial identification. |
| first_indexed | 2024-12-16T09:41:55Z |
| format | Article |
| id | doaj.art-db1f5c0d58a54c5cb67a620b08e6672c |
| institution | Directory Open Access Journal |
| issn | 0972-8988 2231-0916 |
| language | English |
| last_indexed | 2024-12-16T09:41:55Z |
| publishDate | 2017-09-01 |
| publisher | Veterinary World |
| record_format | Article |
| series | Veterinary World |
| spelling | doaj.art-db1f5c0d58a54c5cb67a620b08e6672c2022-12-21T22:36:15ZengVeterinary WorldVeterinary World0972-89882231-09162017-09-011091083109310.14202/vetworld.2017.1083-1093Matrix-assisted laser desorption-ionization-time-of-flight mass spectrometry as a reliable proteomic method for characterization of Escherichia coli and Salmonella isolatesWaleed S. Shell0Mahmoud Lotfy Sayed1Fatma Mohamed Gad Allah2Fatma Elzahraa Mohamed Gamal3Afaf Ahmed Khder4A. A. Samy5Abde Hakam M. Ali6Central Laboratory for Evaluation of Veterinary Biologics Abbasaia, Agriculture Research Center, Cairo, Egypt.Central Laboratory for Evaluation of Veterinary Biologics Abbasaia, Agriculture Research Center, Cairo, Egypt.Central Laboratory for Evaluation of Veterinary Biologics Abbasaia, Agriculture Research Center, Cairo, Egypt.Central Laboratory for Evaluation of Veterinary Biologics Abbasaia, Agriculture Research Center, Cairo, Egypt.Central Laboratory for Evaluation of Veterinary Biologics Abbasaia, Agriculture Research Center, Cairo, Egypt.Department of Microbiology and Immunology, National Research Center, Cairo, Egypt.Central Laboratory for Evaluation of Veterinary Biologics Abbasaia, Agriculture Research Center, Cairo, Egypt.Aim: Identification of pathogenic clinical bacterial isolates is mainly dependent on phenotypic and genotypic characteristics of the microorganisms. These conventional methods are costive, time-consuming, and need special skills and training. An alternative, mass spectral (proteomics) analysis method for identification of clinical bacterial isolates has been recognized as a rapid, reliable, and economical method for identification. This study was aimed to evaluate and compare the performance, sensitivity and reliability of traditional bacteriology, phenotypic methods and matrix-assisted laser desorption-ionization-time-of-flight mass spectrometry (MALDI-TOF MS) in the identification of clinical Escherichia coli and Salmonella isolates recovered from chickens. Materials and Methods: A total of 110 samples (cloacal, liver, spleen, and/or gall bladder) were collected from apparently healthy and diseased chickens showing clinical signs as white chalky diarrhea, pasty vent, and decrease egg production as well as freshly dead chickens which showing postmortem lesions as enlarged liver with congestion and enlarged gall bladder from different poultry farms. Results: Depending on colonial characteristics and morphological characteristics, E. coli and Salmonella isolates were recovered and detected in only 42 and 35 samples, respectively. Biochemical identification using API 20E identification system revealed that the suspected E. coli isolates were 33 out of 42 of colonial and morphological identified E. coli isolates where Salmonella isolates were represented by 26 out of 35 of colonial and morphological identified Salmonella isolates. Serological identification of isolates revealed that the most predominant E. coli serotypes were O1 and O78 while the most predominant Salmonella serotype of Salmonella was Salmonella Pullorum. All E. coli and Salmonella isolates were examined using MALDI-TOF MS. In agreement with traditional identification, MADI-TOF MS identified all clinical bacterial samples with valid scores as E. coli and Salmonella isolates except two E. coli isolates recovered from apparently healthy and diseased birds, respectively, with recovery rate of 93.9% and 2 Salmonella isolates recovered from apparently healthy and dead birds, respectively, with recovery rate of 92.3%. Conclusion: Our study demonstrated that Bruker MALDI-TOF MS Biotyper is a reliable rapid and economic tool for the identification of Gram-negative bacteria especially E. coli and Salmonella which could be used as an alternative diagnostic tool for routine identification and differentiation of clinical isolates in the bacteriological laboratory. MALDI-TOF MS need more validation and verification and more study on the performance of direct colony and extraction methods to detect the most sensitive one and also need using more samples to detect sensitivity, reliability, and performance of this type of bacterial identification.http://www.veterinaryworld.org/Vol.10/September-2017/12.pdfABIBruker DaltonicscolibacillosisEscherichia colimatrix-assisted laser desorption ionization time-of-flight mass spectrometrySalmonellaSalmonella pullorum |
| spellingShingle | Waleed S. Shell Mahmoud Lotfy Sayed Fatma Mohamed Gad Allah Fatma Elzahraa Mohamed Gamal Afaf Ahmed Khder A. A. Samy Abde Hakam M. Ali Matrix-assisted laser desorption-ionization-time-of-flight mass spectrometry as a reliable proteomic method for characterization of Escherichia coli and Salmonella isolates Veterinary World ABI Bruker Daltonics colibacillosis Escherichia coli matrix-assisted laser desorption ionization time-of-flight mass spectrometry Salmonella Salmonella pullorum |
| title | Matrix-assisted laser desorption-ionization-time-of-flight mass spectrometry as a reliable proteomic method for characterization of Escherichia coli and Salmonella isolates |
| title_full | Matrix-assisted laser desorption-ionization-time-of-flight mass spectrometry as a reliable proteomic method for characterization of Escherichia coli and Salmonella isolates |
| title_fullStr | Matrix-assisted laser desorption-ionization-time-of-flight mass spectrometry as a reliable proteomic method for characterization of Escherichia coli and Salmonella isolates |
| title_full_unstemmed | Matrix-assisted laser desorption-ionization-time-of-flight mass spectrometry as a reliable proteomic method for characterization of Escherichia coli and Salmonella isolates |
| title_short | Matrix-assisted laser desorption-ionization-time-of-flight mass spectrometry as a reliable proteomic method for characterization of Escherichia coli and Salmonella isolates |
| title_sort | matrix assisted laser desorption ionization time of flight mass spectrometry as a reliable proteomic method for characterization of escherichia coli and salmonella isolates |
| topic | ABI Bruker Daltonics colibacillosis Escherichia coli matrix-assisted laser desorption ionization time-of-flight mass spectrometry Salmonella Salmonella pullorum |
| url | http://www.veterinaryworld.org/Vol.10/September-2017/12.pdf |
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