Limits...
Matrix-assisted laser desorption ionization-time of flight mass spectrometry based identification of Edwardsiella ictaluri isolated from Vietnamese striped catfish ( Pangasius hypothalamus )

View Article: PubMed Central - PubMed

ABSTRACT

Edwardsiella (E.) ictaluri is a major bacterial pathogen that affects commercially farmed striped catfish (Pangasius hypothalamus) in Vietnam. In a previous study, 19 strains of E. ictaluri collected from striped catfish were biochemically identified with an API-20E system. Here, the same 19 strains were used to assess the ability of matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS; applied using a MALDI Biotyper) to conduct rapid, easy and accurate identification of E. ictaluri. MALDI-TOF MS could directly detect the specific peptide patterns of cultured E. ictaluri colonies with high (> 2.0, indicating species-level identification) scores. MALDI Biotyper 3.0 software revealed that all of the strains examined in this study possessed highly similar peptide peak patterns. In addition, electrophoresis (SDS-PAGE) and subsequent immuno-blotting using a specific chicken antibody (IgY) against E. ictaluri revealed that the isolates had highly similar protein profiles and antigenic banding profiles. The results of this study suggest that E. ictaluri isolated from striped catfish in Vietnam have homologous protein compositions. This is important, because it indicates that MALDI-TOF MS analysis could potentially outperform the conventional methods of identifying E. ictaluri.

No MeSH data available.


Electrophoresis (SDS-PAGE) (A) and Western blot (B) analysis of whole cell lysates from E. ictaluri isolated from striped catfish. Black arrows indicate consistent bands of whole cell lysates.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC5037306&req=5

Figure 2: Electrophoresis (SDS-PAGE) (A) and Western blot (B) analysis of whole cell lysates from E. ictaluri isolated from striped catfish. Black arrows indicate consistent bands of whole cell lysates.

Mentions: SDS-PAGE was used to evaluate the between-strain diversity of the protein profiles. Coomassie Brilliant Blue R-250 staining demonstrated that the resolved protein bands were quite similar in molecular weight at 17–62 kDa, except for 23 kDa, for which some isolates possessed high or low intensity. There were no distinguishable bands < 17 kDa (panel A in Fig. 2).


Matrix-assisted laser desorption ionization-time of flight mass spectrometry based identification of Edwardsiella ictaluri isolated from Vietnamese striped catfish ( Pangasius hypothalamus )
Electrophoresis (SDS-PAGE) (A) and Western blot (B) analysis of whole cell lysates from E. ictaluri isolated from striped catfish. Black arrows indicate consistent bands of whole cell lysates.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC5037306&req=5

Figure 2: Electrophoresis (SDS-PAGE) (A) and Western blot (B) analysis of whole cell lysates from E. ictaluri isolated from striped catfish. Black arrows indicate consistent bands of whole cell lysates.
Mentions: SDS-PAGE was used to evaluate the between-strain diversity of the protein profiles. Coomassie Brilliant Blue R-250 staining demonstrated that the resolved protein bands were quite similar in molecular weight at 17–62 kDa, except for 23 kDa, for which some isolates possessed high or low intensity. There were no distinguishable bands < 17 kDa (panel A in Fig. 2).

View Article: PubMed Central - PubMed

ABSTRACT

Edwardsiella (E.) ictaluri is a major bacterial pathogen that affects commercially farmed striped catfish (Pangasius hypothalamus) in Vietnam. In a previous study, 19 strains of E. ictaluri collected from striped catfish were biochemically identified with an API-20E system. Here, the same 19 strains were used to assess the ability of matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS; applied using a MALDI Biotyper) to conduct rapid, easy and accurate identification of E. ictaluri. MALDI-TOF MS could directly detect the specific peptide patterns of cultured E. ictaluri colonies with high (&gt; 2.0, indicating species-level identification) scores. MALDI Biotyper 3.0 software revealed that all of the strains examined in this study possessed highly similar peptide peak patterns. In addition, electrophoresis (SDS-PAGE) and subsequent immuno-blotting using a specific chicken antibody (IgY) against E. ictaluri revealed that the isolates had highly similar protein profiles and antigenic banding profiles. The results of this study suggest that E. ictaluri isolated from striped catfish in Vietnam have homologous protein compositions. This is important, because it indicates that MALDI-TOF MS analysis could potentially outperform the conventional methods of identifying E. ictaluri.

No MeSH data available.