Comparison of MALDI-TOF MS Biotyper and 16S rDNA sequencing for the identification of Pseudomonas species isolated from fish

Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) was compared to conventional biochemical testing methods and nucleic acid analyses (16S rDNA sequencing, hippurate hydrolysis gene testing, whole genome sequencing [WGS]) for species identification of Campylobacter isolates obtained from chickens ( Gallus gallus domesticus, n = 8), American crows ( Corvus brachyrhynchos, n = 17), a mallard duck ( Anas platyrhynchos, n = 1), and a western scrub-jay ( Aphelocoma californica, n = 1). The test results for all 27 isolates were in 100% agreement between MALDI-TOF MS, the combined results of 16S rDNA sequencing, and the hippurate hydrolysis gene PCR ( p = 0.0027, kappa = 1). Likewise, the identifications derived from WGS from a subset of 14 isolates were in 100% agreement with the MALDI-TOF MS identification. In contrast, biochemical testing misclassified 5 isolates of C. jejuni as C. coli, and 16S rDNA sequencing alone was not able to differentiate between C. coli and C. jejuni for 11 sequences ( p = 0.1573, kappa = 0.0857) when compared to MALDI-TOF MS and WGS. No agreement was observed between MALDI-TOF MS dendrograms and the phylogenetic relationships revealed by rDNA sequencing or WGS. Our results confirm that MALDI-TOF MS is a fast and reliable method for identifying Campylobacter isolates to the species level from wild birds and chickens, but not for elucidating phylogenetic relationships among Campylobacter isolates.

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Contraception: Influence on Vaginal Microbiota and Identification of Vaginal Lactobacilli Using MALDI-TOF MS and 16S rDNA Sequencing

The Open Microbiology Journal ◽

10.2174/1874285801812010218 ◽

2018 ◽

Vol 12 (1) ◽

pp. 218-229 ◽

Cited By ~ 3

Author(s):

Sonia E. Fosch ◽

Cecilia A. Ficoseco ◽

Antonella Marchesi ◽

Silvina Cocucci ◽

Maria E. F. Nader-Macias ◽

...

Keyword(s):

16S Rdna ◽

Oral Contraceptive Pill ◽

Vaginal Microbiota ◽

16S Rdna Sequencing ◽

Contraceptive Methods ◽

Maldi Tof Ms ◽

Maldi Tof ◽

Vaginal Lactobacilli ◽

Rdna Sequencing ◽

Tof Ms

Background:The vaginal microbiome is influenced by a wide variety of factors, including contraceptive methods.Aim:To evaluate the effect of contraceptive methods on vaginal microbiota and to compare MALDI-TOF MS and 16S rDNA sequencing for lactobacilli identification.Patients and Methods:One hundred and one (101) women consulting for birth control were included in a prospective study. Their vaginal content was sampled and analyzed once before they started using the contraceptive method of their choice, and twice after the initiation of contraception, at three months (94/101 women attended) and at six months (89/101 women attended). The relative frequencies of yeasts and trichomonas were analyzed. MALDI-TOF MS and 16S rDNA sequence analysis were applied for the identification of lactobacilli in their vaginal microbiota. The following contraceptive methods were assessed: Combined Oral Contraceptive Pill (COCP), Condom (CON) and The Rhythm Method (RHYT). McNemar’s statistical test was applied.Results:A statistically significant association between COCP and normal microbiota was observed after three months (p< 0.01) and after six months (p< 0.0001), when the vaginal microbiota was modified. At six months, inflammatory reaction was detected in 3/7 women in the CON group, while 6/7 patients using RHYT showed the same state. Yeast colonization increased with the COCP. Identification of lactobacilli by MALDI-TOF MS analysis compared to 16S rDNA sequencing yielded 92.9% concordant results.Lactobacillus gasseriandL. crispatuswere the predominant species.Conclusion:The pattern of vaginal states was significantly modified. Hormone administration apparently corrected the alterations and retained a normal vaginal state. MALDI-TOF MS has the potential of being an accurate tool for the identification of vaginal lactobacilli speciesL. murinuswas for the first time isolated from the vagina.

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Rapid species-level identification of vaginal and oral lactobacilli using MALDI-TOF MS analysis and 16S rDNA sequencing

BMC Microbiology ◽

10.1186/s12866-014-0312-5 ◽

2014 ◽

Vol 14 (1) ◽

Cited By ~ 34

Author(s):

Annette Carola Anderson ◽

Mohamed Sanunu ◽

Christian Schneider ◽

Andreas Clad ◽

Lamprini Karygianni ◽

...

Keyword(s):

16S Rdna ◽

Species Level ◽

16S Rdna Sequencing ◽

Maldi Tof Ms ◽

Maldi Tof ◽

Ms Analysis ◽

Rdna Sequencing ◽

Level Identification ◽

Tof Ms

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Identification of airborne bacteria by 16S rDNA sequencing, MALDI-TOF MS and the MIDI microbial identification system

Aerobiologia ◽

10.1007/s10453-015-9363-9 ◽

2015 ◽

Vol 31 (3) ◽

pp. 271-281 ◽

Cited By ~ 9

Author(s):

Else Marie Fykse ◽

Torbjörn Tjärnhage ◽

Tarmo Humppi ◽

Vilde Sørvik Eggen ◽

Andre Ingebretsen ◽

...

Keyword(s):

16S Rdna ◽

Identification System ◽

Airborne Bacteria ◽

16S Rdna Sequencing ◽

Maldi Tof Ms ◽

Microbial Identification ◽

Maldi Tof ◽

Rdna Sequencing ◽

Microbial Identification System ◽

Tof Ms

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Rapid Identification of Microorganisms Isolated from Pickled Vegetables by MALDI-TOF Mass Spectrometry

Advanced Materials Research ◽

10.4028/www.scientific.net/amr.712-715.494 ◽

2013 ◽

Vol 712-715 ◽

pp. 494-497

Author(s):

Zhi Lei Tan ◽

Yu Qiao Wei ◽

Shuang Liang ◽

Ran Zhang ◽

Miao Liu ◽

...

Keyword(s):

Mass Spectrometry ◽

16S Rdna ◽

Leuconostoc Mesenteroides ◽

16S Rdna Sequencing ◽

Maldi Tof Ms ◽

Maldi Tof ◽

Rdna Sequencing ◽

Pickled Vegetables ◽

Food Bacteria ◽

Tof Ms

Matrix-assisted laser desorption/ionization time-off light mass spectrometry (MALDI-TOF MS) is increasingly used as a microbial diagnostic method for species identification of pathogens. However, MALDI-TOF MS identification of food bacteria was seldom reported. Ten strains isolated from different pickled vegetables were rapid identified by MALDI-TOF MS. The results of MALDI-TOF MS were confirmed by 16S rDNA sequencing method. Different score values in MALDI-TOF MS revealed nineLeuconostoc mesenteroides and oneStaphylococcus.Identification success at the species and genus levels was 90% (9/10) and 100% (10/10), respectively.16S rDNA sequencing results showed that nine stains were identified asLeuconostoc mesenteroides and one wasStaphylococcus saprophyticus.Results obtained demonstrate that MALDI-TOFMS is a promising method for discriminating and identifying food bacteria.

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Influence of High Hydrostatic Pressure on the Identification of Lactobacillus by MALDI-TOF MS- Preliminary Study

Microorganisms ◽

10.3390/microorganisms8060813 ◽

2020 ◽

Vol 8 (6) ◽

pp. 813

Author(s):

Joanna Bucka-Kolendo ◽

Barbara Sokołowska ◽

Stanisław Winiarczyk

Keyword(s):

Mass Spectrum ◽

Hydrostatic Pressure ◽

16S Rdna ◽

High Hydrostatic Pressure ◽

Protein Profile ◽

16S Rdna Sequencing ◽

Maldi Tof Ms ◽

Maldi Tof ◽

Rdna Sequencing ◽

Tof Ms

In the present study, we assessed the ability of MALDI-TOF MS (matrix-assisted laser desorption ionization time-of-flight mass spectrometry) to identify microbial strains subjected to high hydrostatic pressure (HHP) as a stress factor. Protein changes induced by HHP can affect the identification of microorganisms when the identification technique is based on the protein profile. We evaluated two methods, namely MALDI-TOF MS and 16S rDNA sequencing, as a valuable tool to identify Lactobacillus species isolated from spoiled food, juices and beers. The data obtained from the protein mass fingerprint analysis of some of the lactobacilli strains showed differences in unpressured and pressured mass spectrum profiles (MSPs), which influenced the results of the identification. Four out of 13 strains (30%) showed different MSP results for unpressured and pressured samples and these results did not overlap with the 16S rDNA identification results. The 16S rDNA sequencing method revealed that five unpressured strains (38%) and four pressured strains (40%) were identified correctly by MALDI-TOF MS. Both methods showed compatible results in 38% of unpressured strains and in 30% of pressured strains. Stress factors, cultivation methods or the natural environment from which the bacteria were derived can affect their protein profile and thus change the mass spectrum. It is necessary to expand the database with a wide range of mass spectra dedicated to a high-throughput study of the microorganisms derived from different environments.

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Comparison of VITEK2, MALDI-TOF MS, and 16S rDNA sequencing for identification of Myroides odoratus and Myroides odoratimimus

Diagnostic Microbiology and Infectious Disease ◽

10.1016/j.diagmicrobio.2014.02.002 ◽

2014 ◽

Vol 79 (2) ◽

pp. 155-159 ◽

Cited By ~ 25

Author(s):

Percy Schröttner ◽

Wolfram W. Rudolph ◽

Bodo R. Eing ◽

Sebastian Bertram ◽

Florian Gunzer

Keyword(s):

16S Rdna ◽

16S Rdna Sequencing ◽

Maldi Tof Ms ◽

Maldi Tof ◽

Rdna Sequencing ◽

Myroides Odoratimimus ◽

Tof Ms

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MALDI-TOF Mass Spectrometry Fingerprinting Performance Versus 16S rDNA Sequencing to Identify Bacterial Microflora From Seafood Products and Sea Water Samples

Frontiers in Marine Science ◽

10.3389/fmars.2021.650116 ◽

2021 ◽

Vol 8 ◽

Author(s):

Thomas Brauge ◽

Sylvain Trigueros ◽

Arnaud Briet ◽

Sabine Debuiche ◽

Guylaine Leleu ◽

...

Keyword(s):

Mass Spectrometry ◽

16S Rdna ◽

Sea Water ◽

Species Level ◽

16S Rdna Sequencing ◽

Genus Level ◽

Maldi Tof ◽

Rdna Sequencing ◽

Seafood Products ◽

Tof Ms

We evaluated the performance of matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) associated with the Bruker BioTyperTM V7.0.0 database for the identification of 713 bacterial strains isolated from seafood products and sea water samples (ANSES B3PA collection) under culture conditions that may have been significantly different from those used to create the reference spectrum vs. the 16S rDNA sequencing. We identified 78.8% of seafood isolates with 46.7% at the species level (Bruker score above 2) and 21.2% (Bruker score between 1.7 and 2) at the genus level by the two identification methods, except for 3.8% of isolates with a difference of identification between the two methods (Bruker score between 1.7 and 2). There were 41.9% isolates (Bruker score below 1.7) with the identification at the genus level. We identified 94.4% of seafood isolates with 16S rDNA sequencing. The MALDI-TOF allowed a better strain identification to the species level contrary to the 16s rDNA sequencing, which allowed an identification mainly to the genus level. MALDI-TOF MS in association with the Bruker database and 16S rDNA sequencing are powerful tools to identify a wide variety of bacteria from seafood but require further identification by biochemical, molecular technique or other conventional tests.

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