Study of human neutrophil peptides in saliva by matrix‐assisted laser desorption/ionization time‐of‐flight mass spectrometry

Yang, Minghui; Lo, Li‐Hua; Chen, Yi-Hsuan; Shiea, Jentaie; Wu, Pei-Chang; Tyan, Yu‐Chang; Jong, Yuh‐Jyh

doi:10.1002/rcm.4241

Cited by 10 publications

(7 citation statements)

References 70 publications

(61 reference statements)

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“…In addition, we did not include a HNP3 standard in the experimental setting, due to difficulty in finding a highly purified peptide suitable for MS analysis. Still, HNP3 shares the same amino‐acid sequence with HNP1 except one, and the pattern of molecular masses that we detect is comparable with the mass spectra of HNPs previously published 29,30 …”

Section: Discussionsupporting

confidence: 87%

Detection of α‐defensin in synovial fluids by matrix‐assisted laser desorption/ionization time‐of‐flight mass spectrometry as an innovative and cost‐effective assay for improved definition of periprosthetic joint infections

Petrucca

Santino

Gentile

et al. 2020

Rapid Comm Mass Spectrometry

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Rationale:Detection of α-defensins in synovial fluid is gaining more and more interest in the field of correct diagnosis of periprosthetic joint infections (PJIs). At present, they can be assessed by a quantitative enzyme-linked immunosorbent assay which is expensive and time-consuming and by a qualitative lateral flow immunoassay which is rapid but quite expensive and whose clinical sensitivity is debated. Thus, developing an alternative rapid, accurate, and low-cost assay for α-defensins is important to make α-defensins actionable as novel key clinical markers. Methods: Synovial fluid (SF) samples were obtained from 18 patients undergoing revision of primary joint arthroplasty. Of these, eight met the 2013 Musculoskeletal Infection Society (MSIS) criteria for PJIs, the remaining were classified as aseptic failure. Microbiological analysis and Synovasure assays were carried out on all samples. Sample preparation and the matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry (MS) settings were adjusted to detect human neutrophil peptide (HNP)-1, -2 and -3 and to obtain optimal results in term of sensitivity and stability. Results: MALDI-TOF MS was able to detect HNPs in SF from septic patients. No signals for HNPs were detected in SF from aseptic failure. The limits of detection (LOD) were 2.5 and 1.25 μg/mL for HNP-2 and HNP-1, respectively. The turnaround time of the analysis is 20 min, and SF samples are stable at −20 C for up to 3 days.Assay sensitivity, specificity, and positive and negative predictive values (PPV and NPV) were 100% for all parameters. On the same SF samples, the Synovasure assay showed lower sensitivity specificity, and PPV and NPV of 87.5%, 90%, 87.5% and 90%, respectively. Microbiological analysis of SF confirmed the presence of bacteria only in SF MSIS-positive patients. Conclusions:The reported MALDI-TOF MS assay was able to detect and differentiate HNPs in SF samples and showed a slightly better diagnostic accuracy than the Synovasure assay.

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Section: Discussionsupporting

confidence: 87%

Detection of α‐defensin in synovial fluids by matrix‐assisted laser desorption/ionization time‐of‐flight mass spectrometry as an innovative and cost‐effective assay for improved definition of periprosthetic joint infections

Petrucca

Santino

Gentile

et al. 2020

Rapid Comm Mass Spectrometry

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“…The unknown peaks were matched with the peaks obtained from standard Streptococcus mutans BCRC 10793 whose source was from a dental infection site too. 10 These peptide molecules have cationic antimicrobial peptides that are the products of activated and deteriorating neutrophils. Streptococci are important components of the human oral bacterial ora.…”

Section: Resultsmentioning

confidence: 99%

“…HNPs are abundant peptides in saliva. 10 These peptide molecules have cationic antimicrobial peptides that are the products of activated and deteriorating neutrophils. 11 HNPs play very important roles in the innate immune response; they are among the most important components for intrinsic immunity against viruses, fungi and bacteria.…”

Section: Resultsmentioning

confidence: 99%

A brief case study demonstrating the applicability of MALDI mass spectrometry for detecting bacteria in dental samples

Gopal

2015

RSC Adv.

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Samples collected from a gum infection site from a volunteer during surgery were directly analyzed using matrix assisted laser desorption mass spectrometry (MALDI-MS). This paper reports for the first time, the use of MALDI-MS for direct, rapid and sensitive analysis of bacteria from dental samples. The successful detection was due to the extraction of the sample in a microvolume of extractant (water) followed by preconcentration via vortexing coupled with centrifugation.A significant peptide cluster has been identified at the infection site, belonging to Human neutrophil peptides (HNPs). We were able to identify the presence of Streptococcus mutans in the dental samples, using MALDI-MS. This is a bacterium widely reported to be responsible for dental caries at dental infection sites. This approach for the first time reports and validates the unequivocal use of MALDI-MS in dental analysis yielding adequate information in a rapid way (only several minutes) leading to culture free detection of bacteria. This demonstrates that MALDI-MS can be applied as a rapid and sensitive tool for rapid clinical mass spectrometry (RC-MS) in future.

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“…Apart from its physical cleansing and buffering capacity, saliva is also comprised of a number of host immune factors 91,93,235,236 . Recently, families of cationic antimicrobial peptides have been detected in saliva, including the α-defensins (HNP-1-3), the βdefensins (HBD-1-3), the cathelicidin LL-37 and histatins 122,[237][238][239] . These peptides possess broad spectrum antimicrobial activity to various microorganisms.…”

Section: Elisamentioning

confidence: 99%

“…They are expressed from various sources, including salivary gland acini and ducts, oral epithelium and neutrophils. Their levels in saliva are variable among individuals 116,122,237,239,266 .…”

Section: Omentioning

confidence: 99%

The role of salivary antimicrobial peptides in shaping Streptococcus mutans ecology

Phattarataratip¹

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Antimicrobial peptides are among the repertoire of host innate immune defenses. In mucosal immunity, the health-disease balance can be greatly modulated by the interplay between host immune factors and colonized microflora. Microbial ecology within dental plaque is constantly shaped by environmental factors present within the oral cavity. Several antimicrobial peptides are detected in saliva and their bactericidal activities against oral bacteria, including Streptococcus mutans, the primary etiologic agent of dental caries, have been clearly demonstrated. However, the role of these antimicrobial peptides in S .mutans ecology and host caries experience is not welldefined. We hypothesized that various strains of S. mutans possess different inherent susceptibility/resistance profiles to host salivary antimicrobial peptides and that hostspecific quantities of these peptides may influence plaque colonization by particular S. mutans strains. S. mutans strains from subjects with variable caries experience were tested for susceptibility to a panel of antimicrobial peptides, including HNP-1-3, HBD-2-3 and LL-37, revealing that the susceptibilities of S. mutans to these peptides were strain-specific. S. mutans strains from high caries subjects showed greater resistance to these peptides at varying concentrations than those from caries-free subjects. In addition, when combinations of these peptides were tested, they showed either additive or synergistic interaction against S. mutans. Determinations of the salivary levels of these peptides showed that their concentrations were highly variable among subjects with no correlation to host caries experience. However, positive relationships between the salivary concentrations of HNP-encouragement. I am also grateful for kind hospitality and generosity of Dr. Ronald Ettinger and his wife, Sonia, who always provide the warmest support for us, Thai students. Importantly, I would like to thank my mom, dad, sisters, brother and friends in Thailand, including Ann, Oat, Antt and Earth, who have been there for me throughout this long process. I could not have done this without them. Last but not least, I would like to thank the Royal Thai Government Scholarship for providing me the financial support for my graduate study and giving me the opportunity to study here in Iowa.

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Study of human neutrophil peptides in saliva by matrix‐assisted laser desorption/ionization time‐of‐flight mass spectrometry

Cited by 10 publications

References 70 publications

Detection of α‐defensin in synovial fluids by matrix‐assisted laser desorption/ionization time‐of‐flight mass spectrometry as an innovative and cost‐effective assay for improved definition of periprosthetic joint infections

Detection of α‐defensin in synovial fluids by matrix‐assisted laser desorption/ionization time‐of‐flight mass spectrometry as an innovative and cost‐effective assay for improved definition of periprosthetic joint infections

A brief case study demonstrating the applicability of MALDI mass spectrometry for detecting bacteria in dental samples

The role of salivary antimicrobial peptides in shaping Streptococcus mutans ecology

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