MALDI-TOF MS performance compared to direct examination, culture, and 16S rDNA PCR for the rapid diagnosis of bone and joint infections

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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MALDI-TOF MS for rapid diagnosis of Anaerobiospirillum succiniciproducens, an unusual causative agent of bacteraemia in humans. Two case reports and literature review

Anaerobe ◽

10.1016/j.anaerobe.2018.12.003 ◽

2019 ◽

Vol 55 ◽

pp. 130-135 ◽

Cited By ~ 3

Author(s):

Marina Alguacil-Guillen ◽

Luis Ramos-Ruperto ◽

Juan Carlos Ramos Ramos ◽

Angel Robles-Marhuenda ◽

Julio García-Rodriguez ◽

...

Keyword(s):

Literature Review ◽

Causative Agent ◽

Case Reports ◽

Rapid Diagnosis ◽

Maldi Tof Ms ◽

Anaerobiospirillum Succiniciproducens ◽

Maldi Tof ◽

Tof Ms

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

Microbial Pathogenesis ◽

10.1016/j.micpath.2019.04.024 ◽

2019 ◽

Vol 132 ◽

pp. 313-318 ◽

Cited By ~ 3

Author(s):

Miroslava Kačániová ◽

Alīna Klūga ◽

Attila Kántor ◽

Juraj Medo ◽

Jana Žiarovská ◽

...

Keyword(s):

16S Rdna ◽

16S Rdna Sequencing ◽

Maldi Tof Ms ◽

Maldi Tof ◽

Rdna Sequencing ◽

Pseudomonas Species ◽

Tof Ms

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Porównanie metod identyfikacji bakterii Lactobacillus

Zywnosc Nauka Technologia Jakosc/Food Science Technology Quality ◽

10.15193/zntj/2021/127/377 ◽

2021 ◽

Vol 127 (2) ◽

pp. 49-60

Author(s):

Joanna Bucka-Kolendo ◽

Barbara Sokołowska

Keyword(s):

16S Rdna ◽

Maldi Tof Ms ◽

Maldi Tof ◽

Tof Ms

Szerokie zastosowanie bakterii kwasu mlekowego (LAB) w różnych gałęziach przemysłu spożywczego, w biotechnologii i w medycynie powoduje, że właściwa identyfikacja i ocena ich zróżnicowania wewnątrzgatunkowego jest bardzo istotna. Dobór odpowiednich technik molekularnych analizy powinien uwzględniać dużą dokładność, powtarzalności i typowalność metody. Celem pracy była ocena możliwości różnicowania 12 szczepów Lactobacillus przy użyciu metod powszechnie stosowanych w laboratoriach: sekwencjonowania genu 16S rDNA, genu pheS oraz MALDI-TOF MS. Na podstawie otrzymanych wyników analiz stwierdzono, że gen pheS w badanych szczepach charakteryzował się wysokim poziomem homologii (98 %) i niską siłą dyskryminacyjną. W dwóch niezależnych analizach MALDI-TOF MS uzyskano taki sam wynik dla 10 szczepów: siedmiu – L. brevis (DSM 6235, 102, 103, 489, 863, 975, 3/16/1), dwóch – L. plantarum (1178, 133) i jednego – L. curvatus 557. Po przeprowadzeniu analizy sekwencjonowania genu 16S rDNA wyniki potwierdziły się natomiast tylko w odniesieniu do pięciu szczepów (DSM 6235, 3/16/1 i 489) z dwunastu przebadanych. Porównanie wyników było podstawą do wnioskowania, że dla wybranych szczepów LAB największą wartość różnicującą miała analiza bazująca na sekwencjonowaniu genu 16S rDNA.

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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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Rapid direct detection of pathogens for diagnosis of joint infections by MALDI-TOF MS after liquid enrichment in the BacT/Alert blood culture system

PLoS ONE ◽

10.1371/journal.pone.0243790 ◽

2020 ◽

Vol 15 (12) ◽

pp. e0243790

Author(s):

Christine Noll ◽

Azadda Nasruddin-Yekta ◽

Pia Sternisek ◽

Michael Weig ◽

Uwe Groß ◽

...

Keyword(s):

Synovial Fluid ◽

Blood Culture ◽

Direct Detection ◽

Direct Identification ◽

Maldi Tof Ms ◽

Maldi Tof ◽

Flight Mass Spectrometry ◽

Joint Infections ◽

Solid Media ◽

Tof Ms

Pathogen identification is a critical step during diagnosis of infectious diseases. Matrix-Assisted Laser Desorption/Ionization Time-Of-Flight mass spectrometry (MALDI-TOF-MS) has become the gold standard for identification of microorganisms cultured on solid media in microbiology laboratories. Direct identification of microbes from liquid specimen, circumventing the need for the additional overnight cultivation step, has been successfully established for blood culture, urine and liquor. Here, we evaluate the ability of MALDI-TOF MS for direct identification of pathogens in synovial fluid after liquid enrichment in BacT/Alert blood culture bottles. Influence of synovial specimen quality on direct species identification with the MALDI BioTyper/Sepsityper was tested with samples inoculated from pretested native synovia with concomitant inoculation of blood or pus, or highly viscous fluid. Here, we achieved >90% concordance with culture on solid medium, and only mixed-species samples posed significant problems. Performance in routine diagnostics was tested prospectively on bottles inoculated by treating physicians on ward. There, we achieved >70% concordance with culture on solid media. The major contributors to test failure were the absence of a measurable mass signal and mixed-specimen samples. The Sepsityper workflow worked well on samples derived from BacT/Alert blood culture bottles inoculated with synovial fluid, giving concordant results to identification from solid media. Host remnant material in the inoculum, such as blood or pus, had no detrimental effect on identification score values of the BioTyper system after processing with the Sepsityper workflow, and neither had the initial viscosity of the synovial sample.

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