Reference Standards for C-Peptide in Korean Population: A Korean Endocrine Hormone Reference Standard Data Center Study.

Background: The Korean Endocrine Hormone Reference Standard Data Center (KEHRS DC) has created reference standards (RSs) for endocrine hormones since 2020. This study is the first of its kind, wherein the KEHRS DC established RSs for serum Cpeptide levels in a healthy Korean population. Methods: Healthy Korean adults were recruited from May 2021 to September 2023. After excluding participants according to our criteria, serum samples were collected; each participant could then choose between fasting glucose only or fasting glucose plus an oral glucose tolerance test (OGTT). If their sample showed high glucose (≥100 mg/dL) or hemoglobin A1c (HbA1c) (≥5.70%), their C-peptide levels were excluded from analyzing the RSs. Results: A total of 1,532 participants were recruited; however, only the data of 1,050 participants were analyzed after excluding those whose samples showed hyperglycemia or high HbA1c. Post-30-minute OGTT data from 342 subjects and post-120-minute OGTT data from 351 subjects were used. The means±2 standard deviations and expanded uncertainties of fasting, post-30-minute and 120-minute OGTT C-peptide levels were 1.26±0.82 and 0.34-3.18, 4.74±3.57 and 1.14-8.33, and 4.85±3.58 and 1.25-8.34 ng/mL, respectively. Serum C-peptide levels correlated with obesity, serum glucose levels, and HbA1c levels. Conclusion: The RSs for serum C-peptide levels established in this study are expected to be useful in both clinical and related fields.

Culture-Free Quantification of Bacteria Using Digital Fluorescence Imaging in a Tunable Magnetic Capturing Cartridge for Onsite Food Testing.

Accurate, onsite detection of pathogenic bacteria from food matrices is required to rapidly respond to pathogen outbreaks. However, accurately detecting whole-cell bacteria in large sample volumes without an enrichment step remains a challenge. Therefore, bacterial samples must be concentrated, identified, and quantified. We developed a tunable magnetic capturing cartridge (TMCC) and combined it with a portable digital fluorescence reader for quick, onsite, quantitative detection of Staphylococcus aureus. The TMCC platform integrates an absorption pad impregnated with water-soluble polyvinyl alcohol (PVA) with an injection-molded polycarbonate (PC) plate that has a hard magnet on its back and an acrylonitrile-butadiene-styrene case. An S. aureus-specific antibody conjugated with magnetic nanoparticles was used to concentrate bacteria from a large-volume sample and capture bacteria within the TMCC. The retention time for capturing bacteria on the TMCC was adjusted by controlling the concentration and volume of the PVA solution. Concentrated bacterial samples bound to target-specific aptamer probes conjugated with quantum dots were loaded into the TMCC for a controlled time, followed by attachment of the bacteria to the PC plate and removal of unbound aptamer probes with wash buffer. The captured bacteria were quantified using a digital fluorescence reader equipped with an embedded program that automatically counts fluorescently tagged bacteria. The bacterial count made using the TMCC was comparable to a standard plate count (R2 = 0.9898), with assay sensitivity and specificity of 94.3 and 100%, respectively.

Sensitive electrochemical immunosensor to detect prohibitin 2, a potential blood cancer biomarker.

Blood cancers are difficult to cure completely and frequently show a poor prognosis. Recently, prohibitin 2 (PHB2) has been shown to be a potential biomarker for blood cancers. Sandwich ELISA can be used as a reference method for quantitative analysis of PHB2; however, ELISA can be challenging for early diagnosis and continuous monitoring method due to the need for large sample volumes (25 µL <), technical expertise, complex procedure, relative high cost, and non-portability. Thus, this study developed a sensitive and time efficient electrochemical immunosensor for detecting PHB2 from a blood cancer patient. It is a simple and portable platform consisting of a disposable electrode and blood sample volume of 4 µL. The sensor uses a gold nanostructured electrode and square wave voltammetry (SWV) measurement of a horseradish peroxidase (HRP) label to amplify the electrochemical signal. The immunosensor could quantitatively detect PHB2 with high sensitivity (limit of detection [LoD] = 0.04 ng/mL) and satisfactory reproducibility (relative standard deviation [RSD] <5.2%). The sensor achieved an LoD of 0.63 ng/mL with satisfactory recovery (89.1-104.7%) and reproducibility (RSD <6.4%) with PHB2 spiked into white blood cell (WBC) lysates. When the sensor was compared to a reference ELISA to determine the PHB2 concentrations in WBC lysate samples from healthy patients and those with blood cancer, the correlation coefficient (R2) was 0.996. A 3.3-fold difference was detected in the measured PHB2 concentration between blood cancer patients and healthy individuals. Accordingly, this study suggests a sensitive and accurate analytical method for quantitatively detecting the PHB2 in blood samples.

Rapid bacteria-detection platform based on magnetophoretic concentration, dielectrophoretic separation, and impedimetric detection.

Most biosensors employ small sample quantities (less than 100 µL) for bacteria detection, thereby resulting in inaccurate low-concentration measurements. Detection performed using small sample volumes with low bacteria concentration may produce false-negative results. Therefore, sample pretreatment plays a critical role in accurate bacteria detection. This paper presents an impedimetric bacteria-detection sensor integrated with bacteria concentration and separation devices for rapid bacteria detection. Post conjugation using magnetic particles (MPs), the MP-conjugated bacteria (MP/Bac) are concentrated via magnetophoresis by a factor exceeding 100. In addition, MP/Bac are separated from MPs via dielectrophoresis to prevent occurrence of signal errors caused by MPs not conjugated with bacteria. Subsequently, concentrated MP/Bac are captured on a sensor electrode, and bacteria concentration is detected by measuring signal changes caused by the impedance difference between bacteria and the medium. The performance of the proposed bacteria-detection device was evaluated using a 5-mL homogenized cabbage sample injected with Staphylococcus aureus at 30 mL/h flow rate. The observed signal change was measured for 10 min using a sample with a concentration of 5-5 × 103 CFU/mL and was found to be approximately 0.34 mV at 50 CFU/mL; the limit of detection was 36 CFU/mL. These results confirm that the proposed device can detect low bacteria concentrations in food samples.

Marker Pen Device with Concentration Gradient Nib for Antibiotic Susceptibility Testing.

BACKGROUND: Pen-based devices have emerged as useful tools for measuring pH and glucose, and for fabricating microchannels and microarrays. Pen-based devices take advantage of flexible patterning, inexpensive costs, and small volumes, thereby saving time and increasing efficiency. We have developed a gradient nib marker pen device that generated simultaneously different antibiotic concentrations in bacteria antibiotic susceptibility testing (AST). METHODS: The device can deposit on the target surface with the antibiotic gradient. The designed polyester fiber nibs are a highly uniform porosity with unidirectional orientation and produce a visible gradient pattern. RESULTS: We have demonstrated and quantitatively analyzed bacterial growth after antibiotic marking. The antibiotic marking produces an inhibition zone of bacterial growth. The inhibition zones of bacterial growth are captured and converted to 8-bit grayscale images, and then quantified by gray values using the Image J program. A profile of the inhibition zone showed different gray values in response to bacterial viability. CONCLUSION: The gradient nib marker pen device can be used to determine the quantitative antibiotic concentration based on the relationship between gray values and bacterial density conveniently without requiring a series of dilution tubes, including nutrient medium, and diversely diluted antibiotics.