Identification of Lysinibacillus fusiformis Isolated from Cosmetic Samples Using MALDI-TOF MS and 16S rRNA Sequencing Methods

2018 ◽  
Vol 101 (6) ◽  
pp. 1757-1762 ◽  
Author(s):  
Irshad M Sulaiman ◽  
Ying-Hsin Hsieh ◽  
Emily Jacobs ◽  
Nancy Miranda ◽  
Steven Simpson ◽  
...  
Keyword(s):  
16S Rrna ◽  
16S Rrna Sequencing ◽  
Gram Positive ◽  
Maldi Tof Ms ◽  
Lysinibacillus Fusiformis ◽  
Maldi Tof ◽  
Microbiological Methods ◽  
Rrna Sequencing ◽  
Level Identification ◽  
Tof Ms

Abstract Background: Lysinibacillus fusiformis is a Gram-positive, rod-shaped bacterium that can cause tropical ulcers, severe sepsis, and respiratory illnesses in humans. Objective: In this study, we analyzed cosmetic samples for the presence of human pathogenic microorganisms. Methods: Five unopened jars of exfoliating cream were examined initially by microbiological methods. Afterward, matrix-assisted laser desorption ionization-time-of-flight (MALDI-TOF) MS and 16S ribosomal RNA (rRNA) sequencing techniques were applied to characterize the recovered isolates. Results: Of the eight recovered Gram-positive bacterial subs, the VITEK® MS could provide genus-level identification to five subs and species-level identification to two subs (L. fusiformis with a 99.9% confidence value); one sub was unidentified. Subsequently, the deoxyriboneucleic acid sequencing of the 16S rRNA gene was done on an ABI 3500XL Genetic Analyzer for the confirmation of species identification. An analysis of sequencing data revealed a complete absence of genetic variation among the eight subs sequenced at this locus and confirmed the eight bacterial subs to be L. fusiformis, as their respective 16S rRNA sequences were identical to the available sequence in public domain (GenBank accession No. KU179364). Conclusions: Our results suggest that the VITEK MS and the 16S rRNA sequencing can be used for the identification of human pathogenic bacteria of public health importance. Highlights: We characterized eight isolates of Lysinibacillus spp. from cosmetics by MALDI-TOF MS and 16S rRNA sequence analyses.

scholarly journals MALDI-TOF MS versus 16S rRNA sequencing: Minor discrepancy between tools in identification of Bacteroides isolates

2018 ◽  
Vol 65 (2) ◽  
pp. 173-181
Author(s):  
Károly Péter Sárvári ◽  
József Sóki ◽  
Miklós Iván ◽  
Cecilia Miszti ◽  
Krisztina Latkóczy ◽  
...  
Keyword(s):  
16S Rrna ◽  
16S Rrna Sequencing ◽  
Maldi Tof Ms ◽  
Maldi Tof ◽  
Rrna Sequencing ◽  
Tof Ms

Identification and classification of seafood-borne pathogenic and spoilage bacteria: 16S rRNA sequencing versus MALDI-TOF MS fingerprinting

2013 ◽  
Vol 34 (6) ◽  
pp. 877-887 ◽  
Author(s):  
Karola Böhme ◽  
Inmaculada C. Fernández-No ◽  
Manuel Pazos ◽  
José M. Gallardo ◽  
Jorge Barros-Velázquez ◽  
...  
Keyword(s):  
16S Rrna ◽  
16S Rrna Sequencing ◽  
Maldi Tof Ms ◽  
Maldi Tof ◽  
Spoilage Bacteria ◽  
Rrna Sequencing ◽  
Tof Ms

Anaerobic blood culture isolates in a Norwegian university hospital: identification by MALDI-TOF MS vs 16S rRNA sequencing and antimicrobial susceptibility profiles

Apmis ◽  
2015 ◽  
Vol 123 (9) ◽  
pp. 749-758 ◽  
Author(s):  
Nina Handal ◽  
Silje Bakken Jørgensen ◽  
Hege Smith Tunsjø ◽  
Bjørn Odd Johnsen ◽  
Truls Michael Leegaard
Keyword(s):  
Blood Culture ◽  
16S Rrna ◽  
Antimicrobial Susceptibility ◽  
16S Rrna Sequencing ◽  
University Hospital ◽  
Maldi Tof Ms ◽  
Maldi Tof ◽  
Rrna Sequencing ◽  
Susceptibility Profiles ◽  
Tof Ms

scholarly journals Multicenter Evaluation of Rapid BACpro® II for the Accurate Identification of Microorganisms Directly from Blood Cultures Using MALDI-TOF MS

Diagnostics ◽  
2021 ◽  
Vol 11 (12) ◽  
pp. 2251
Author(s):  
Marina Oviaño ◽  
André Ingebretsen ◽  
Anne K. Steffensen ◽  
Antony Croxatto ◽  
Guy Prod’hom ◽  
...  
Keyword(s):  
Blood Cultures ◽  
Species Level ◽  
Correct Identification ◽  
Gram Positive ◽  
Gram Negative ◽  
Maldi Tof Ms ◽  
Gram Positive Bacteria ◽  
Maldi Tof ◽  
Level Identification ◽  
Tof Ms

The identification of microorganisms directly from blood cultures using MALDI-TOF MS has been shown to be the most impacting application of this methodology. In this study, a novel commercial method was evaluated in four clinical microbiology laboratories. Positive blood culture samples (n = 801) were processed using a rapid BACpro® II kit and then compared with the routine gold standard. A subset of monomicrobial BCs (n = 560) were analyzed in parallel with a Sepsityper® Kit (Bruker Daltonics, Bremen, Germany) and compared with the rapid BACpro® II kit. In addition, this kit was also compared with two different in-house methods. Overall, 80.0% of the monomicrobial isolates (609/761; 95% CI 71.5–88.5) were correctly identified by the rapid BACpro® II kit at the species level (92.3% of the Gram negative and 72.4% of the Gram positive bacteria). The comparison with the Sepsityper® Kit showed that the rapid BACpro® II kit generated higher rates of correct species-level identification for all categories (p > 0.0001), except for yeasts identified with score values > 1.7. It also proved superior to the ammonium chloride method (p > 0.0001), but the differential centrifugation method allowed for higher rates of correct identification for Gram negative bacteria (p > 0.1). The percentage of accurate species-level identification of Gram positive bacteria was particularly noteworthy in comparison with other commercial and in-house methods.

scholarly journals Evaluation of the rapidBACpro® II kit for the rapid identification of microorganisms directly from blood cultures using MALDI-TOF MS

2021 ◽  
Author(s):  
Marina Oviaño ◽  
André Ingebretsen ◽  
Anne K Steffensen ◽  
Antony Croxatto ◽  
Guy Prod’hom ◽  
...  
Keyword(s):  
Blood Cultures ◽  
Species Level ◽  
Correct Identification ◽  
Gram Positive ◽  
Gram Negative ◽  
Maldi Tof Ms ◽  
Gram Positive Bacteria ◽  
Maldi Tof ◽  
Level Identification ◽  
Tof Ms

AbstractObjectivesIdentification of microorganisms directly from blood cultures (BCs) using MALDI-TOF MS has shown to be the application with most impact in this methodology. In this study, a novel commercial method, the rapidBACpro® II, was evaluated in four clinical microbiology laboratories.MethodsPositive blood culture samples (n=801) were processed using the rapidBACpro® II kit and then compared with routine gold standard. A subset of monomicrobial BCs (n=560) were analyzed in parallel with the Sepsityper® kit (Bruker Daltonics, Bremen, Germany) and compared with the rapidBACpro® II kit. In addition, the rapidBACpro® II kit was also compared with two different in-house methods.ResultsOverall, 80.0% of the monomicrobial isolates (609/761) were correctly identified by the rapidBACpro® II kit at the species level (92.3% of the Gram negative and 72.4% of the Gram positive bacteria). The comparison with the Sepsityper® kit yielded higher rates of correct species-level identification provided by the rapidBACpro® II kit for all categories (p>0.0001) except for yeasts identified with score values >1.7. It also proved superior to the ammonium chloride method (p>0.0001) but the differential centrifugation method allowed higher rates of correct identification for Gram negative bacteria (p>0.1).ConclusionsThe rapidBACpro® II kit allowed a high rate of microorganisms correctly identified. The percentage of accurate species-level identification of Gram positive bacteria was particularly noteworthy in comparison with other commercial and in-house methods. This fact was especially interesting in the case of Staphylococcus sp. and Streptococcus sp. in order to elucidate their clinical impact, for example in device-associated bacteremia.

scholarly journals Matrix-assisted Laser Desorption Ionization-Time-of-Flight Mass Spectrometry (MALDI-TOF MS) as a Reliable Tool to Identify Species of Catalase-negative Gram-positive Cocci not Belonging to the Streptococcus Genus

2016 ◽  
Vol 10 (1) ◽  
pp. 202-208 ◽  
Author(s):  
Marisa Almuzara ◽  
Claudia Barberis ◽  
Viviana Rojas Velázquez ◽  
Maria Soledad Ramirez ◽  
Angela Famiglietti ◽  
...  
Keyword(s):  
Extraction Methods ◽  
Species Level ◽  
Ionization Time ◽  
Maldi Tof Ms ◽  
Genus Level ◽  
Maldi Tof ◽  
Flight Mass Spectrometry ◽  
Level Identification ◽  
Gram Positive Cocci ◽  
Tof Ms

Objective:To evaluate the performance of matrix-assisted laser desorption ionization-time-of-flight mass spectrometry (MALDI-TOF MS) by using 190 Catalase-negative Gram-Positive Cocci (GPC) clinical isolates.Methods:All isolates were identified by conventional phenotypic tests following the proposed scheme by Ruoff and Christensen and MALDI-TOF MS (Bruker Daltonics, BD, Bremen, Germany). Two different extraction methods (direct transfer formic acid method on spot and ethanol formic acid extraction method) and different cut-offs for genus/specie level identification were used. The score cut-offs recommended by the manufacturer (≥ 2.000 for species-level, 1.700 to 1.999 for genus level and <1.700 no reliable identification) and lower cut-off scores (≥1.500 for genus level, ≥ 1.700 for species-level and score <1.500 no reliable identification) were considered for identification. A minimum difference of 10% between the top and next closest score was required for a different genus or species.MALDI-TOF MS identification was considered correct when the result obtained from MS database agreed with the phenotypic identification result.When both methods gave discordant results, the 16S rDNA orsodAgenes sequencing was considered as the gold standard identification method. The results obtained by MS concordant with genes sequencing, although discordant with conventional phenotyping, were considered correct. MS results discordant with 16S orsodA identification were considered incorrect.Results:Using the score cut-offs recommended by the manufacturer, 97.37% and 81.05% were correctly identified to genus and species level, respectively. On the other hand, using lower cut-off scores for identification, 97.89% and 94.21% isolates were correctly identified to genus and species level respectively by MALDI-TOF MS and no significant differences between the results obtained with two extraction methods were obtained.Conclusion:The results obtained suggest that MALDI-TOF MS has the potential of being an accurate tool for Catalase-negative GPC identification even for those species with difficult diagnosis asHelcococcus,Abiotrophia,Granulicatella, among others. Nevertheless, expansion of the library, especially including more strains with different spectra on the same species might overcome potential “intraspecies” variability problems. Moreover, a decrease of the identification scores for species and genus-level identification must be considered since it may improve the MALDI-TOF MS accuracy.

scholarly journals Discrimination of Streptococcus equi subsp. equi and Streptococcus equi subsp. zooepidemicus using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry

2017 ◽  
Vol 29 (5) ◽  
pp. 622-627 ◽  
Author(s):  
Rinosh J. Mani ◽  
Anil J. Thachil ◽  
Akhilesh Ramachandran
Keyword(s):  
Laser Desorption Ionization ◽  
Biochemical System ◽  
Ionization Time ◽  
Maldi Tof Ms ◽  
Maldi Tof ◽  
Flight Mass Spectrometry ◽  
Subspecies Level ◽  
Streptococcus Equi ◽  
Level Identification ◽  
Tof Ms

Accurate and timely identification of infectious etiologies is of great significance in veterinary microbiology, especially for critical diseases such as strangles, a highly contagious disease of horses caused by Streptococcus equi subsp. equi. We evaluated a matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) platform for use in species- and subspecies-level identification of S. equi isolates from horses and compared it with an automated biochemical system. We used 25 clinical isolates each of S. equi subsp. equi and S. equi subsp. zooepidemicus. Using the MALDI-TOF MS platform, it was possible to correctly identify all 50 isolates to the species level. Unique mass peaks were identified in the bacterial peptide mass spectra generated by MALDI-TOF MS, which can be used for accurate subspecies-level identification of S. equi. Mass peaks (mass/charge, m/ z) 6,751.9 ± 1.4 (mean ± standard deviation) and 5,958.1 ± 1.3 were found to be unique to S. equi subsp. equi and S. equi subsp. zooepidemicus, respectively. The automated biochemical system correctly identified 47 of 50 of the isolates to the species level as S. equi, whereas at the subspecies level, 24 of 25 S. equi subsp. equi isolates and 22 of 25 S. equi subsp. zooepidemicus isolates were correctly identified. Our results indicate that MALDI-TOF MS can be used for accurate species- and subspecies-level identification of S. equi.

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