scholarly journals Mass Spectrometry Based-Proteomic Analysis of Anisakis spp.: A Preliminary Study towards a New Diagnostic Tool

Genes ◽  
2020 ◽  
Vol 11 (6) ◽  
pp. 693 ◽  
Author(s):  
Valeria Marzano ◽  
Stefania Pane ◽  
Gianluca Foglietta ◽  
Stefano Levi Mortera ◽  
Pamela Vernocchi ◽  
...  
Keyword(s):  
Diagnostic Tool ◽  
Protein Extraction ◽  
Shotgun Proteomics ◽  
Diagnostic Tools ◽  
Routine Practice ◽  
Maldi Tof Ms ◽  
Maldi Tof ◽  
Proteomics Approach ◽  
Anisakis Spp ◽  
Tof Ms

Anisakiasis is nowadays a well-known infection, mainly caused by the accidental ingestion of Anisakis larvae, following the consumption of raw or undercooked fishes and cephalopods. Due to the similarity of symptoms with those of common gastrointestinal disorders, this infection is often underestimated, and the need for new specific diagnostic tools is becoming crucial. Given the remarkable impact that MALDI–TOF MS biotyping had in the last decade in clinical routine practice for the recognition of bacterial and fungi strains, a similar scenario could be foreseen for the identification of parasites, such as nematodes. In this work, a MALDI–TOF MS profiling of Anisakis proteome was pursued with a view to constructing a first spectral library for the diagnosis of Anisakis infections. At the same time, a shotgun proteomics approach by LC–ESI–MS/MS was performed on the two main fractions obtained from protein extraction, to evaluate the protein species enriched by the protocol. A set of MALDI–TOF MS signals associated with proteins originating in the ribosomal fraction of the nematode extract was selected as a potential diagnostic tool for the identification of Anisakis spp.

scholarly journals Candida auris Clinical Isolates from South Korea: Identification, Antifungal Susceptibility, and Genotyping

2019 ◽  
Vol 57 (4) ◽  
Author(s):  
Yong Jun Kwon ◽  
Jong Hee Shin ◽  
Seung A Byun ◽  
Min Ji Choi ◽  
Eun Jeong Won ◽  
...  
Keyword(s):  
Antifungal Susceptibility ◽  
Diagnostic Tools ◽  
Candida Auris ◽  
Content Type ◽  
Maldi Tof Ms ◽  
Maldi Tof ◽  
Fluconazole Resistant ◽  
Vitek 2 ◽  
Vitek Ms ◽  
Tof Ms

ABSTRACT Candida auris is an emerging worldwide fungal pathogen. Over the past 20 years, 61 patient isolates of C. auris (4 blood and 57 ear) have been obtained from 13 hospitals in Korea. Here, we reanalyzed those molecularly identified isolates using two matrix-assisted laser desorption ionization–time of flight mass spectrometry (MALDI-TOF MS) systems, including Biotyper and Vitek MS, followed by antifungal susceptibility testing, sequencing of the ERG11 gene, and genotyping. With a research-use-only (RUO) library, 83.6% and 93.4% of the isolates were correctly identified by Biotyper and Vitek MS, respectively. Using an in vitro diagnostic (IVD) library of Vitek MS, 96.7% of the isolates were correctly identified. Fluconazole-resistant isolates made up 62.3% of the isolates, while echinocandin- or multidrug-resistant isolates were not found. Excellent essential (within two dilutions, 96.7%) and categorical agreements (93.4%) between the Clinical and Laboratory Standards Institute (CLSI) and Vitek 2 (AST-YS07 card) methods were observed for fluconazole. Sequencing ERG11 for all 61 isolates revealed that only 3 fluconazole-resistant isolates showed the Erg11p amino acid substitution K143R. All 61 isolates showed identical multilocus sequence typing (MLST). Pulsed-field gel electrophoresis (PFGE) analyses revealed that both blood and ear isolates had the same or similar patterns. These results show that MALDI-TOF MS and Vitek 2 antifungal susceptibility systems can be reliable diagnostic tools for testing C. auris isolates from Korean hospitals. The Erg11p mutation was seldom found among Korean isolates of C. auris, and multidrug resistance was not found. Both MLST and PFGE analyses suggest that these isolates are genetically similar.

scholarly journals Identification of Nocardia species using Autof MS 1000 and Bruker MALDI-TOF MS systems: A comparison

2021 ◽  
Author(s):  
Bing Ma ◽  
Yunqi Tian ◽  
Yungang Han ◽  
Lifang Ban ◽  
Junwen Yang ◽  
...  
Keyword(s):  
Species Identification ◽  
Protein Extraction ◽  
Reference Method ◽  
Invasive Disease ◽  
Species Level ◽  
Maldi Tof Ms ◽  
Maldi Tof ◽  
Flight Mass Spectrometry ◽  
Significant Difference ◽  
Tof Ms

ABSTRACTNocardia is an important cause of clinically invasive disease, but for most clinical laboratories, identification of these isolates to the species level is challenging. Recently, matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS) has been widely used for identification of most bacterial and fungal isolates. In this multicenter study, we evaluated the identification of Nocardia isolates using Autof MS1000 and Bruker Biotyper. A total of 86 non-duplicate Nocardia isolates from 7 hospital laboratories were evaluated. Further, we carried out sequence analysis of 16S rRNA, gyrB, secA1, hsp65, and rpoB genes as a reference method for Nocardia species identification. The 86 isolates were directly spotted on the target plate and plate protein extraction was performed. Data were analyzed by SPSS 19.0. In total, 72 (83.7%) strains (score ≥ 9.0) and 70 (81.4%) strains (score ≥ 2.0) were correctly identified by the Autof MS1000 and Bruker Biotyper systems, respectively, at the species level. There was no significant difference (P > 0.05) between the two systems using the same protein extraction method. In conclusion, the Autof MS 1000 and Bruker MALDI-TOF systems showed no difference in identification of Nocardia spp. to the species level and could meet the most important clinical requirement for species identification.

scholarly journals Usefulness of MALDI-TOF MS as a Diagnostic Tool for the Identification of Streptococcus Species Recovered from Clinical Specimens of Pigs

PLoS ONE ◽  
2017 ◽  
Vol 12 (1) ◽  
pp. e0170784 ◽  
Author(s):  
Marta Pérez-Sancho ◽  
Ana I. Vela ◽  
Teresa García-Seco ◽  
Sergio González ◽  
Lucas Domínguez ◽  
...  
Keyword(s):  
Diagnostic Tool ◽  
Clinical Specimens ◽  
Maldi Tof Ms ◽  
Streptococcus Species ◽  
Maldi Tof ◽  
Tof Ms

scholarly journals Evaluation of Two Protein Extraction Protocols Based on Freezing and Mechanical Disruption for Identifying Nontuberculous Mycobacteria by Matrix-Assisted Laser Desorption Ionization–Time of Flight Mass Spectrometry from Liquid and Solid Cultures

2018 ◽  
Vol 56 (4) ◽  
Author(s):  
David Rodriguez-Temporal ◽  
Daniel Perez-Risco ◽  
Eduardo A. Struzka ◽  
Mireia Mas ◽  
Fernando Alcaide
Keyword(s):  
Protein Extraction ◽  
Nontuberculous Mycobacteria ◽  
Laser Desorption Ionization ◽  
Liquid Media ◽  
Ionization Time ◽  
Maldi Tof Ms ◽  
Maldi Tof ◽  
Flight Mass Spectrometry ◽  
Mechanical Disruption ◽  
Tof Ms

ABSTRACTMatrix-assisted laser desorption ionization–time of flight mass spectrometry (MALDI-TOF MS) has proved to be a useful diagnostic method for identifying conventional bacteria. In the case of mycobacteria, a good protein extraction protocol is essential in order to obtain reliable identification results. To date, no such protocol has been definitively established. The aim of this study was to compare the manufacturer's recommended protein extraction protocol (protocol A) with two novel protocols (protocols B and C), which apply different freezing temperatures and mechanical disruption times using an automatic tissue homogenizer. A total of 302 clinical isolates, comprising 41 nontuberculous mycobacteria (NTM) species, were grown in parallel on solid and liquid media and analyzed: 174 isolates were slow-growing mycobacteria (SGM) and 128 isolates were rapid-growing mycobacteria (RGM). Overall, MALDI-TOF MS identified a higher number of NTM isolates from solid than from liquid media, especially with protocol C (83.4 and 68.2%, respectively;P< 0.05). From solid media, this protein extraction method identified 57.9 and 3.9% more isolates than protocols A (P< 0.001) and B (P< 0.05), respectively. In the case of liquid media, protocol C identified 49.7 and 6.3% more isolates than protocols A and B, respectively (P< 0.001). With regard to the growth rate, MALDI-TOF MS identified more RGM isolates than SGM isolates in all of the protocols studied. In conclusion, the application of freezing and automatic tissue homogenizer improved protein extraction of NTM and boosted identification rates. Consequently, MALDI-TOF MS, which is a cheap and simple method, could be a helpful tool for identifying NTM species in clinical laboratories.

MALDI-ToF/MS as a diagnostic tool for the confirmation of sulfur mustard exposure†**

2001 ◽  
Vol 20 (S1) ◽  
pp. S193-S197 ◽  
Author(s):  
Elvis O. Price ◽  
J. Richard Smith ◽  
Connie R. Clark ◽  
John J. Schlager ◽  
Ming L. Shih
Keyword(s):  
Diagnostic Tool ◽  
Sulfur Mustard ◽  
Maldi Tof Ms ◽  
Maldi Tof ◽  
Tof Ms

MALDI-TOF MS as evolving cancer diagnostic tool: A review

2014 ◽  
Vol 95 ◽  
pp. 245-255 ◽  
Author(s):  
Miguel Angel Merlos Rodrigo ◽  
Ondrej Zitka ◽  
Sona Krizkova ◽  
Amitava Moulick ◽  
Vojtech Adam ◽  
...  
Keyword(s):  
Diagnostic Tool ◽  
Maldi Tof Ms ◽  
Cancer Diagnostic ◽  
Maldi Tof ◽  
Tof Ms

Rapid one-step protein extraction method for the identification of mycobacteria using MALDI-TOF MS

2019 ◽  
Vol 94 (4) ◽  
pp. 355-360 ◽  
Author(s):  
Suwatchareeporn Rotcheewaphan ◽  
Jamie K. Lemon ◽  
Uma U. Desai ◽  
Christina M. Henderson ◽  
Adrian M. Zelazny
Keyword(s):  
Extraction Method ◽  
Protein Extraction ◽  
Maldi Tof Ms ◽  
Maldi Tof ◽  
One Step ◽  
Tof Ms

scholarly journals MALDI-TOF MS and point of care are disruptive diagnostic tools in Africa

2018 ◽  
Vol 26 ◽  
pp. S83-S88 ◽  
Author(s):  
E. Chabriere ◽  
H. Bassène ◽  
M. Drancourt ◽  
C. Sokhna
Keyword(s):  
Point Of Care ◽  
Diagnostic Tools ◽  
Maldi Tof Ms ◽  
Maldi Tof ◽  
Tof Ms

A novel protein extraction method for identification of mycobacteria using MALDI-ToF MS

2015 ◽  
Vol 119 ◽  
pp. 1-3 ◽  
Author(s):  
La'Tonzia L. Adams ◽  
Parichat Salee ◽  
Kim Dionne ◽  
Karen Carroll ◽  
Nicole Parrish
Keyword(s):  
Extraction Method ◽  
Protein Extraction ◽  
Maldi Tof Ms ◽  
Maldi Tof ◽  
Tof Ms ◽  
Novel Protein
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