Analysis Of Saliva Research Articles

THE USE OF MULTI-OMICS TO IDENTIFY PREDICTIVE FACTORS FOR FOOD REACTIONS

<h3>Introduction</h3> The pathophysiology of IgE mediated food reactions (FR) is multifactorial. This multi-omic analyses utilized host and microbial factors to better understand the immunological origins of FR's. <h3>Methods</h3> This longitudinal cohort study included 129 term infants, identified as FR (n=27) or non-FR (n=92) using the Infant Feeding Practices-II survey (confirmed with medical records). Saliva was collected at 6 months for multi-omic analysis of cytokines, mRNA, microRNAs, and the microbiome. <h3>Results</h3> None of the salivary cytokines (CXCL10, IL-8, IL-6, IL-18) displayed differences between groups. However, the Th1/Th2 ratio (IL-8/IL-6) was nominally higher (d = 0.56, p = 0.043, FDR = 0.30) among infants with a food reaction. One mRNA (STAT6, p = 0.039, FDR = 0.17, d = 0.045), two miRNAs (let-7e-5p, p = 0.031, FDR = 0.12, d = 0.51; miR-151a-5p, p = 0.041, FDR = 0.13, d = 0.52), and one bacterial phylum (Cyanobacteria, p = 0.024, FDR = 0.19, d = 0.72) displayed nominal differences between 27 infants with food reactions (FR) and 92 peers with no food reactions (NFR). All four omics features were higher in infants with FR. A logistic regression model employing maternal/infant traits and environmental exposures and four multi-omic features (IL-8/IL-6, miR-151a-5p, STAT6, Cyanobacteria) accounted for an additional 31% of the variance between groups (p = 0.15, AUC = 0.83, sensitivity = 97%, specificity = 48%). <h3>Conclusion</h3> Multi-omic analysis of saliva may enhance understanding of key biological factors that contribute towards developing immune mediated food reactions.

Saliva and plasma steroidome in mare during reproductive stages: A GC-MS/MS study

Precision livestock farming using omics approach to acquire precise and real-time data can help farmers in individual animal management and decision making. Since steroid hormones play a key role for the regulation of reproductive functions, reproduction management could be improved by characterizing the steroidome during reproductive stages. Moreover, saliva collection is a non-invasive, painless, inexpensive and easy sampling method. Thus, this prospective study proposes a steroidomic analysis in mare saliva during reproductive stages, that could help to identify potential biomarkers to accurately detect their reproductive stage in a welfare friendly production system, for real-time decision making at the individual animal level. Correlation between saliva and plasma steroidome was also investigated. Saliva and blood samples from 6 mares were collected in anestrus, in the follicular phase 3 days, 2 days and 1 day before ovulation and the day when ovulation was detected, in the luteal phase 6 days after ovulation and in gestation 18 days after ovulation and insemination. Steroidome analysis was performed by gas chromatography coupled to tandem mass spectrometry (GC-MS/MS). We characterized 25 and 36 steroids in saliva and plasma respectively. Pregnenolone concentrations in saliva during gestation were significantly higher than during anestrus or follicular phase and tended to be higher than during luteal phase. Most of the 5α-reduced metabolites of progesterone showed higher salivary concentrations during the luteal phase 6 days post-ovulation and during gestation 18 days post-ovulation compared to anestrus and follicular phase. These steroids could be potential salivary biomarkers of the reproductive stage of the mare. Further studies with a greater number of animals are in progress to confirm the reliability of these potential candidate biomarkers and to develop field-friendly assays.

Comparative Analysis of Saliva and Plasma Proteins Patterns in Pregnant Cows-Preliminary Studies.

Simple SummaryOne of the most crucial topics about cattle breeding is pregnancy. During this state, there are many changes in protein expression and abundance. These changes find reflection not only in plasma protein patterns but also in saliva, which is easier to obtain than blood. The aim of this study was the analysis of plasma and salivary protein profiles in pregnant cows in order to search for valuable markers of pregnancy status. In this study, the presence of apolipoproteins possibly related to bovine pregnancy was confirmed both in plasma and saliva. This means that saliva can be considered a good source of information about the condition of the organism, including during pregnancy. It is possible that the comparison of salivary and plasma proteomes can be a helpful tool to assess the pregnancy status of cattle, and can be useful for developing rapid tests from saliva.Pregnancy is a physiological state that can be described, from a biochemical point of view, using protein patterns. The present study focused on the comparison of protein patterns between the saliva and plasma of pregnant cows to search for possible markers which are present both in plasma and saliva. Saliva and plasma were collected from healthy, pregnant (3–4 months) and non-pregnant (C; n = 4) cows aged between 4 and 8 years (P; n = 8) from the same farm. Biological material was analyzed using 2D electrophoresis and MS identification. Among identified spots, there were those which could be related to pregnancy (e.g., apolipoproteins I and II in all examined matrices or transforming growth factor-beta-induced protein ig-h3 in albumin-free plasma) as well as those which are responsible for regulating of cellular processes (e.g., pyruvate kinase and aspartate aminotransferase in all examined matrices, or lactate dehydrogenase, phosphoglycerate kinase, and NADH dehydrogenase in plasma). Further identification of common spots and those only specific to saliva as well as the comparison between other periods of pregnancy are necessary; it is already clear that saliva can be considered a valuable diagnostic matrix containing potential markers of physiological and pathological status.

Saliva Analysis of pH and Antioxidant Capacity in Adult Obstructive Sleep Apnea Patients.

Background: Obstructive sleep apnea syndrome (OSAS) may be associated with and activates the stress response system, and variation in the physiological antioxidant capacity of body fluids. Our aim was to evaluate the variation of pH and antioxidant capacity on the saliva of obstructive sleep apnea subjects (OG) compared to a control group (CG). Method: Fifty subjects with moderate/severe OSAS were recruited in Tor Vergata Hospital and compared with 20 healthy subjects CG. The buffer and the antioxidant capacity of the samples were quantified measuring the pH and the percentage of total salivary antioxidant capacity (%TAC), which refers to the reduced glutathione salivary concentration (GSH). Moreover, the protein concentration and the gelatinolytic activity of metalloproteinases were quantified. Results: The data showed that the pH value is slightly more alkaline in OSAS subjects; however, it is not directly related to the severity of OSAS. The %TAC was found to be significantly reduced by 86.2% in the OG. Proteins of saliva from the OG were found 1.5 times more concentrated than in the healthy sample. The gelatinolytic activity of metalloproteinases of healthy and OSA did not show statistically significant changes. Conclusions: The salivary samples from OSAS compared to CG show an alteration of the oxidative state, the pH buffering power, and protein concentrations, inducing conditions that can easily evolve chronic gingivitis. Further investigations are necessary to evaluate the feasibility of using salivary fluid for the early diagnosis of oral or systemic problems in OSAS subjects.

A rapid UHPLC-QDa method for quantification of human salivary amino acid profiles

Abnormal salivary amino acid (AA) levels may indicate dysfunction of the body. Being noninvasive, sampling easily and cost-effective of saliva, a rapid, precise and simple analysis method has become very important for quantitative salivary AA profiles. After one-step to precipitate protein, the resultant extraction was derived with 6-aminoquinolyl-N-hydroxysuccinimidyl carbamate (AQC) within 10 min. Quantitation of AA profile was achieved within 6 min in a single run by ultra-high performance liquid chromatography coupled with a single quadrupole mass spectrometry (UHPLC-QDA detector). The method was validated with acceptable accuracy ranging from 80.33 % to 122.31 %, appropriate linearity with the coefficient (R2) more than 0.991, good intra- and inter-day precision, repeatability and stability (RSD < 15 %). The recoveries at three different spiked concentrations ranged over 79.18 %–125.36 % while the matrix effect was from −19.86 % to 11.95 %. This simple, rapid and robust method was successfully applied to quantify human salivary 30 amino acids, in which the levels of taurine, γ-aminobutyric acid, methionine and tryptophan in healthy people were close to the LOQs. Besides, the levels of histidine and cystine were not able to be measured due to their relatively high LOQs, which were considered as the limitations of this developed method.

Salivary proteomic analysis in asymptomatic and symptomatic SARS-CoV-2 infection: Innate immunity, taste perception and FABP5 proteins make the difference

Background and aimSARS-CoV-2 infection spawns from an asymptomatic condition to a fatal disease. Age, comorbidities, and several blood biomarkers are associated with infection outcome. We searched for biomarkers by untargeted and targeted proteomic analysis of saliva, a source of viral particles and host proteins. MethodsSaliva samples from 19 asymptomatic and 16 symptomatic SARS-CoV-2 infected subjects, and 20 controls were analyzed by LC-MS/MS for untargeted peptidomic (flow through of 10 kDa filter) and proteomic (trypsin digestion of filter retained proteins) profiling. ResultsPeptides from 53 salivary proteins were identified. ADF was detected only in controls, while IL1RA only in infected subjects. PRPs, DSC2, FABP5, his-1, IL1RA, PRH1, STATH, SMR3B, ANXA1, MUC7, ACTN4, IGKV1-33 and TGM3 were significantly different between asymptomatic and symptomatic subjects. Retained proteins were 117, being 11 highly different between asymptomatic and symptomatic (fold change ≥2 or ≤−2). After validation by LC-MS/MS-SRM (selected reaction monitoring analysis), the most significant discriminant proteins at PCA were IL1RA, CYSTB, S100A8, S100A9, CA6, and FABP5. ConclusionsThe differentially abundant proteins involved in innate immunity (S100 proteins), taste (CA6 and cystatins), and viral binding to the host (FABP5), appear to be of interest for use as potential biomarkers and drugs targets.

Influence of different sample preparation strategies on hypothesis-driven shotgun proteomic analysis of human saliva

Abstract This research aimed to find an efficient and repeatable bottom-up proteolytic strategy to process the unstimulated human saliva. The focus is on monitoring immune system activation via the cytokine and interleukin signaling pathways. Carbohydrate metabolism is also being studied as a possible trigger of inflammation and joint damage in the context of the diagnostic procedure of temporomandibular joint disorder. The preparation of clean peptide mixtures for liquid chromatography–mass spectrometry analysis was performed considering different aspects of sample preparation: the filter-aided sample preparation (FASP) with different loadings of salivary proteins, the unfractionated saliva, amylase-depleted, and amylase-enriched salivary fractions. To optimize the efficiency of the FASP method, the protocols with the digestion in the presence of 80% acetonitrile and one-step digestion in the presence of 80% acetonitrile were used, omitting protein reduction and alkylation. The digestion procedures were repeated in the standard in-solution mode. Alternatively, the temperature of 24 and 37°C was examined during the trypsin digestion. DyNet analysis of the hierarchical networks of Gene Ontology terms corresponding to each sample preparation method for the bottom-up assay revealed the wide variability in protein properties. The method can easily be tailored to the specific samples and groups of proteins to be examined.

Simple and rapid method for 336 multiresidual pesticide analysis in saliva, determination of their chemical stabilities, and biomonitoring of farmers

Simultaneous multiresidual pesticide analysis of saliva samples was performed using scaled-down QuEChERS extraction with LC-MS/MS and GC-MS/MS. The optimum extraction procedure using acidified acetonitrile was applicable to 336 pesticides (287 for LC-MS/MS and 49 for GC-MS/MS). To determine pesticide multiresidues in saliva, 100 μL of the sample was extracted with 200 μL of 0.1% formic acid in acetonitrile, and the initial extract was partitioned with 40 mg of MgSO4 and 10 mg of NaCl. The organic supernatants (120 μL) were then mixed with acetonitrile (30 μL) for matrix-matching (4:1, v/v), and the final extract solution was injected into the LC-MS/MS (4 μL) and GC-MS/MS (2 μL) systems. The established analytical method showed a good LOQs between 5 and 25 ng/mL with reliable accuracy/precision values and recovery results (50–140%) for the target pesticides. Under the two different storage conditions, most of the analytes did not undergo chemical changes in the saliva samples, whereas some pesticides were more stable in freeze-thaw processes than those left at room temperature. Biomonitoring of farmers (ten mixers and ten sprayers) was successfully applied using the validated method, and two carbamates (fenobucarb and propamocarb) were determined at trace concentrations (12.5–675.0 ng/mL from 11 positively detected samples).

Method to Diagnose Diabetes through Saliva

A. Background A prevalent long-term condition that has been linked to salivary amylase levels is diabetes mellitus (DM). Recently, salivary amylase diagnostics have been linked to DM. The metabolic alterations that the DM population goes through have an impact on their salivary parameters. Saliva is a special fluid that is necessary for the mouth cavity to operate normally. Saliva collection is less intrusive, simpler, and technically insensitive than blood collection, making diagnosis by saliva analysis potentially helpful. The primary benefit of this approach is that it is a quick diagnostic tool. Establishing a conservative approach to measuring blood sugar in place of venous blood samples can be aided by examining the link between blood glucose levels and its concentration in saliva. Depending on how saliva was collected under fasting or non-fasting settings, there were variations in salivary amylase levels. The type of diabetes, the kind of insulin treatment, or the level of glycemic control determines the variations in salivary amylase. B. Methodology Human saliva is an organic liquid vital created by the salivary organs. Saliva contains a few biomarkers which makes it valuable for multiplexed measures that are being created for point-of-care gadgets, quick tests, or for centralised clinical research facility tasks. The most significant perception is that proteins present in the blood are comparably present in saliva. Saliva-based diagnostics can likewise help in contriving early treatment systems. Salivary glucose focuses were seen as uniquely different in diabetes mellitus. This is on the grounds that the salivary organs act as a filter of blood glucose that are adjusted by hormonal or neural regulation. Since parts of saliva are derived from blood, the grouping of biochemical and immunological parts estimated in saliva could reflect blood levels. This prompt expanded emission of glucose from the ductal cells of the salivary organ, consequently expanding the glucose content in saliva. Salivary glucose can be used as an elective technique for diabetes and as a general evaluating apparatus for pre-diabetes and undiagnosed diabetes features the significant parts of saliva (harmless) and blood (intrusive). There have been a few reports showing biochemical changes in the saliva of diabetic patients. These modifications are related to salivary egg whites, amylase, support limit, electrolytes, glucose focus, IgA, IgG, IgM, lysozyme, peroxidase, and complete protein levels. Sampling, transport, and capacity of saliva are exceptionally straightforward when compared with blood. The entire mouth saliva is a salivary fluid and every one of the additional items incorporate cells from the mouth, nasal bodily fluid, blood from gum bruises, food flotsam and jetsam, and microbiota. For exact proteomic examination, the mucosal transudate furthermore, gingival crevicular are better impressions of the blood constituents. The materials and strategies used to gather saliva might impact the precision of testing. Prompt refrigeration at 4 degrees C would protect tests in the event that freezing is absurd yet support at this temperature ought to be no longer than 2 h prior to freezing at - 20 degrees C. Storage methodology and time from the collection principally influence the examination of the biochemical factors described by temperature instability and microbial development. C. Results &amp; Conclusions Contrasted with the blood, saliva contains a comparable assortment of constituents that can be utilised for the diagnosis of diabetes mellitus. Salivary glucose levels can be analysed as a noninvasive symptomatic. In addition to biochemical and metabolomic analysis, a paper strip-based non-invasive glucose biosensor was effectively created for salivary examination to analyse diabetes. Saliva protein profiling could be an alluring chance to analyse and screen diabetes in future. Therefore, salivary diagnostics has developed into a sophisticated discipline and fills in as a region of the bigger field of molecular diagnostics, presently perceived as a vital participant in biomedical, fundamental, and clinical examination.

Measuring the Content of Endocannabinoid-Like Compounds in Biological Fluids: A Critical Overview of Sample Preparation Methodologies.

Different mass spectrometric techniques have been used over the past decade to quantify endocannabinoids (eCBs) and related lipids. Even with the level of molecular fingerprinting accuracy of an instrument like the most advanced triple quadrupole mass spectrometer, if one is not getting the most optimized sample to the detector in a way that this improved technology can be of use, then advancements can be stymied. Here, our focus is on review and discussion of sample preparation methodologies used to isolate the eCB anandamide and its close congeners N-acyl ethanolamines and structural congeners (i.e., lipo amino acids, lipoamines, N-acyl amides) in biological fluids. Most of our focus will be on the analysis of these lipids in plasma/serum, but we will also discuss how the same techniques can be used for the analysis of saliva and breast milk.

High-level gonosomal mosaicism for a pathogenic non-coding CNV deletion of the lung-specific FOXF1 enhancer in an unaffected mother of an infant with ACDMPV.

Alveolar capillary dysplasia with misalignment of pulmonary veins (ACDMPV) results from haploinsufficiency of the mesenchymal transcription factor FOXF1 gene. To date, only one case of an ACDMPV-causative CNV deletion inherited from a very-low level somatic mosaic mother has been reported. Clinical, histopathological, and molecular studies, including whole genome sequencing, chromosomal microarray analysis, qPCR, and Sanger sequencing, followed by in vitro fertilization (IVF) with preimplantation genetic testing (PGT) were used to study a family with a deceased neonate with ACDMPV. A pathogenic CNV deletion of the lung-specific FOXF1 enhancer in the proband was found to be inherited from an unaffected mother, 36% mosaic for this deletion in her peripheral blood cells. The qPCR analyses of saliva, buccal cells, urine, nail, and hair samples revealed 19%, 18%, 15%, 19%, and 27% variant allele fraction, respectively, indicating a high recurrence risk. Grandparental studies revealed that the deletion arose on the mother's paternal chromosome 16. PGT studies revealed 44% embryos with the deletion, reflecting high-level germline mosaicism. Our data further demonstrate the importance of parental testing in ACDMPV families and reproductive usefulness of IVF with PGT in families with high-level parental gonosomal mosaicism.

Evaluation of salivary visfatin, leptin and oxidative stress markers in obese and non-obese periodontitis patients.

Objectives: The aim of this study is to evaluate leptin, visfatin, nitric oxide(NO), total oxidant capacity(TOC) and antioxidant capacity(TAOC) levels in saliva at obese and non-obese periodontitis patients. Materials and Methods: Total of 78 individuals were included in study, 20 obese periodontal healthy(Group I), 18 obese periodontitis(Group II), 20 non-obese periodontal healthy(Group III) and 20 non-obese periodontitis(Group IV). Periodontal clinical parameters, body mass index(BMI) and waist circumference measurements were recorded.ELISA and Griess methods were used for biochemical analysis of saliva. Results: It was determined that leptin and TAOC levels were higher in periodontal healthy groups than in the periodontitis groups; visfatin, NO and TOC levels were higher in periodontitis groups than periodontally healthy groups(P

Saliva analysis in children with active caries before and after dental treatment

Background: The amount and quality of saliva play important roles in maintaining an intraoral bacterial balance. The quality of saliva is defined by its buffering capacity, viscosity, pH and protein content. The amount of saliva is usually related to the flow rate. Purpose: This study aimed to compare the flow rate, pH, viscosity and buffering capacity of saliva as well as plaque formation in children before and after dental treatment. Methods: Saliva samples were taken from paediatric patients before their treatments and one month after their dental treatments had ended, and these saliva samples were then analysed. For each sample analysis, the GC Saliva-Check Buffer kit (GC Corporation, Tokyo, Japan) was used to evaluate buffering capacity, pH and flow rate, and the GC Saliva-Check Mutans kit (GC Corporation, Tokyo, Japan) was used for the determination of Streptococcus Mutans. GC Tri Plaque ID gel (GC Corporation, Tokyo, Japan) was applied to evaluate plaque maturation. Results: The pre-treatment buffering capacity, pH and viscosity sample values were found to be significantly lower than the post-treatment values (p&lt;0.05). No statistically significant difference was determined in the amount of saliva pre- and post-treatment (p&gt;0.05). When examining plaque maturation, it was determined that all of the post-treatment plaque was pink. Conclusion: This study showed that the pH, viscosity and buffering capacity of saliva had increased significantly post-treatment and that the formation of plaque had decreased in children with active caries after all their dental treatments had been completed.

The Academy of Breastfeeding Medicine27th Annual International MeetingBaltimore, MarylandSeptember 15–18, 2022

The Academy of Breastfeeding Medicine27^th Annual International MeetingBaltimore, MarylandSeptember 15–18, 2022

A novel electrochemical strategy for NT-proBNP detection using IMFET for monitoring heart failure by saliva analysis

Heart failure (HF) is a chronic cardiovascular disease that represents main cause of mortality worldwide, particularly for elderly. N-terminal pro-brain natriuretic peptide (NT-proBNP) was identified as the gold standard biomarker for HF diagnosis and therapy monitoring. Presently, saliva analysis represents an emerging and powerful tool for clinical applications and electrochemical immunosensors have shown their potential in Healthcare applications as selective and reliable systems for detecting clinical biomarkers. This work presents the detection of NT-proBNP in saliva samples by an immunologically modified Field effect Transistor (IMFET). TESUD ((11-triethoxysilyl) undecanal) was used as cross-linker to immobilise anti-NT-proBNP antibody onto the device. Our IMFET that was then tested in different matrices (e.g. phosphate buffered saline (PBS), artificial saliva and human saliva) using electrochemical impedance spectroscopy (EIS), and it resulted selective to NT-proBNP with good sensitivity (detection limit of 0.02 pg/mL) and a wide linear range (0.02–1 pg/mL and 0.5–20 pg/mL). Finally, NT-proBNP concentration in ten saliva samples was determined by performing the standard addition method. An enzyme-linked immunosorbent assay was used for confirming IMFET results, highlighting both IMFET accuracy (analyte recovery of 99 ± 8%) and precision (coefficient of variation always <10%), and supporting the suitability of the device for determining salivary NT-proBNP.

Development of analytical methods to study the salivary metabolome: impact of the sampling.

Advances in metabolomics have allowed the identification and characterization of saliva metabolites that can be used as biomarkers. However, discrepancies can be noted with the content of the same biomarker being increased or decreased for a given disease. Differences in the way saliva is collected, stored, and/or treated could cause these discrepancies. Indeed, there is no standardized method for saliva sampling and analysis. In this work, two chromatographic modes were used, i.e., RP-LC and HILIC both coupled to MS used in positive and negative ionization modes. The analytical conditions were optimized with a mixture of 90 compounds naturally present in saliva, representative of the wide range of molecular mass and polarity of salivary metabolites and being described as having a differential expression in various pathologies. These four methods were applied to the analysis of saliva samples collected by spitting, aspiration, or Salivette® with or without prior rinsing of the mouth. Rinsing had an effect on some metabolite concentrations. As it can induce an additional parameter of variability to the sampling, it seems therefore preferable to use methods without rinsing while effects of these parameters on the metabolites are investigated. Saliva obtained by spitting and aspiration gave statistically equivalent results for 84% of the metabolites studied. Conversely, Salivette® gave different results since the majority of the metabolites chosen for the study were not quantified in the samples. The Salivette® does not seem therefore to be a suitable sampling method for an untargeted analysis of the salivary metabolome, unlike aspiration and spitting.

FTIR spectroscopy as a point of care diagnostic tool for diabetes and periodontitis: A saliva analysis approach

Diabetes mellitus (DM) is a general term for heterogeneous metabolic disorders whose main characteristic is chronic hyperglycemia. Considering that conventional diagnostic methods are currently unable of early DM detection and the number of diabetic patients has been increasing worldwide, there is a clinical need for novel diagnostic approaches. Fourier-transform infrared (FT-IR) spectroscopy technique an alternative non-invasive diagnostic method for real-time evaluation of biofluids such as saliva. This study aims evaluate the feasibility of diagnosing diabetes and periodontitis through saliva samples based on their FT-IR spectra. Our first collection and spectral analysis of samples was a pilot study and comprised a total of 23 patients, 2 healthy, 9 with diabetes and 12 with diabetes and periodontitis. By using weighted KNN as classifier, we have obtained an area under the Receiver Operating Characteristic curve of 0.92 and 0.95 when considering the diabetes or diabetes + periodontitis groups as positive groups, respectively.

502 The genomic and phenotypic landscape of ichthyosis: An analysis of 1000 kindreds

Objective: Clear genotype-phenotype associations in ichthyosis have been difficult to establish. We aimed to expand the genotypic and phenotypic spectra of ichthyosis and delineate genotype-phenotype associations. Design: We recruited an international group of individuals with ichthyosis, including participants of all ages, races, and ethnicities. Genetic analysis of saliva or blood, a phenotyping questionnaire, and standardized clinical photographs were obtained. Results: Among 1000 unrelated individuals, 266 novel variants in 32 genes were identified.

A System-on-Board Integrated Multi-analyte PoC Biosensor for Combined Analysis of Saliva and Exhaled Breath.

The need for oral health monitoring Point of Care (PoC) systems is ever growing. This is effectively highlighted by the ongoing COVID-19 pandemic where the lack of rapid PoC testing has placed an unsustainable burden on centralized laboratory testing. Urgent development has furthered pathogenic nucleic acid and antibody detection in oral samples throat swabs, but without corresponding advancements in biochemical monitoring through oral biosensing. We have recently reported two novel biosensor technologies for detection of high impact hormones: cortisol in saliva by organic electrolyte gated FETs (OEGFETs), and 8-isoprostane in exhaled breath condensate (EBC) using molecularly imprinted electroimpedance spectroscopy biosensors (MIP EIS). In this work, we report a first stage integration of the two biosensors - previously bench-top proven - with a miniaturized semi-hermetically sealed soft-fluidic enclosure, onto a low-power (<300 mW) customized printed circuit board. Our findings established comparable detection thresholds for the miniaturized board-based configuration and a lab-based test setup, and their ability to characterize, calibrate, and operate these small footprint biosensors. Testing with the 8-isoprostane EBC MIP EIS biosensors showed the system-on-board had an effective frequency range of 100-100kHz, comparable to lab bench impedance analyzers. Despite internal impedance increases of 210%, the expected data features are present in the impedance graphs collected with the PCB. The system-on-board experiments using OEGFET aptasensor showed a predictable behavior and comparable sensor detection range and resolution using unadulterated supernatant and serial dilutions of cortisol over a range of 273 μM to 2.73pM. The portable, multi-analyte oral biosensor is a promising prototype for future packaging and clinical validation.

A surface‐enhanced Raman sensor for trace identification and analysis of high‐priority drugs of abuse with portable and handheld Raman devices

AbstractSurface‐enhanced Raman spectroscopy (SERS) is a rapidly emerging technology that offers a fast, extremely sensitive, and quantitative approach to trace chemical detection. Such a technique incorporated into a portable device is attractive to law enforcement and emergency room personnel for rapid accurate on‐site screening of illicit and abused drugs. Toward this, we are developing an analyzer in a hand‐held unit, based on SERS that is integrated with a capillary sensor embedded with activated gold nanoparticles in a porous glass matrix. In this preliminary study, we have used this gold sol–gel capillary to measure aqueous solutions of 14 high‐priority drugs to define sensitivity (lowest measured concentration [LMC] and estimated limit of detection [LOD]), determine concentration dependence and quantification capabilities by constructing calibration curves, examine effects of pH at 3, 7, and 11 on SERS measurements, and perform multicomponent analysis of fentanyl laced drug mixtures and spectral identification on our portable and handheld units. Of particular significance, fentanyl was detected with an LMC of 1 ng/ml and estimated LOD at 0.11 ng/ml, while other representative drugs such as cocaine and phenylcyclidine produced LMCs and LODs at 5.0 and 0.77 ng/ml, and 10.0 and 0.62 ng/ml, respectively. These sub‐nanogram per milliliter detection limits are comparable or lower than previously reported and confirm that this gold sol–gel has the potential to meet the sensitivity requirements for saliva analysis. Most importantly, these sensors can be manufactured easily and cheaply and when integrated with our portable Raman units produce high‐quality spectra and accurate identification within 1.2 s.