Clinical chemistry最新文献

{"title":"A-088 Clinically Significant Errors Using the Diazyme Total Bile Acid Assay on the Roche c502 Analyzer: Investigating a Result Error Identifies a Novel Large-Scale Testing Error Caused by Reagent Carryover","authors":"L M Leonard, M A Nicklas, D R Block, N A Baumann","doi":"10.1093/clinchem/hvae106.087","DOIUrl":"https://doi.org/10.1093/clinchem/hvae106.087","url":null,"abstract":"Background Thoroughly investigating single patient result errors may identify systemic, large-scale testing errors. The laboratory observed an event where a revision to a Total Bile Acid (TBA, Diazyme Laboratories, Inc.) patient result following a 22S quality control (QC) failure displayed a larger than expected difference (52 to 5mcmol/L). TBA is primarily used for diagnosis and monitoring of intrahepatic cholestasis of pregnancy with results >10mcmol/L considered elevated. Retrospective review of result revisions following QC failures (3/15/2017-5/30/2023) revealed that 50%(20/40) yielded result differences of >10mcmol/L(>20SD based on assay imprecision). The aim of this study was to apply a systematic approach to i) estimate the rate of TBA errors, ii) ensure accurate result reporting during investigation period, iii) perform root cause analysis (RCA), and iv) determine corrective and preventative action(s). Methods Residual samples from TBA testing (6/24-7/5/2023) were retested(n=158), differences >+/-3.0mcmol/L or 15% were confirmed on an alternate analyzer, and reports were revised. Automated repeat testing of TBA samples >10mcmol/L was operationalized 7/6/2023. Initial and repeat TBA results were compared(n=448) and results differing by >+/-20% were remeasured on an alternate analyzer. RCA was conducted using a fishbone diagram. NaOH reagent probe washes were implemented for TBA and the error rate was re-assessed. Assays run prior to TBA samples with errors (n=15) were identified. To assess reagent carryover a residual serum pool (TBA ∼5mcmol/L) was aliquoted into 5 tubes in a sample rack. The first sample in the rack was programmed to run an assay suspected of causing carryover followed by 4 TBA measurements. Mean±standard deviation(SD) TBA concentrations were calculated. TBA was also measured in the liquid reagent for amylase, lipase, acetylcholinesterase (ACE) and fructosamine. Results Initial TBA retesting yielded no errors when initial TBA was ≤10mcmol/L(n=51). For samples with TBA >10mcmol/L(n=107), 9(8.4%) had differences exceeding criteria with 8/9 being revised to ≤10mcmol/L. Analysis of automated patient repeat data showed a 3.8%(17/448) error rate when initial TBA >10mcmol/L. After NaOH reagent probe washes were implemented, the error rate decreased to 0.7%(3/448). Assays run directly before an erroneously high TBA result included: lactate, fructosamine, soluble transferrin receptor(STFR), lipase, and ACE. A serum TBA pool(mean±SD=5.1±0.4 mcmol/L,n=15) measured 37.6±1.2mcmol/L(n=3) and 6.7±0.7mcmol/L(n=3) after lipase and fructosamine, respectively. No other assays demonstrated carryover. TBA in lipase reagent compartment B and C was 653 and 649mcmol/L, respectively, and 653mcmol/L in fructosamine reagent compartment B. Conclusions A single patient TBA result revision was investigated and led to identification of reagent carryover causing erroneously high TBA results on the","PeriodicalId":10690,"journal":{"name":"Clinical chemistry","volume":"9 1","pages":""},"PeriodicalIF":9.3,"publicationDate":"2024-10-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142369123","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"A-083 Performance evaluation of Sentinel Diagnostics’ ACE assay on the Abbott Alinity c system and ARCHITECT c16000 System","authors":"S Brambilla, C Assalini, E Frana, R Lucini, D Novati","doi":"10.1093/clinchem/hvae106.082","DOIUrl":"https://doi.org/10.1093/clinchem/hvae106.082","url":null,"abstract":"Background The analytical performances of the Angiotensin Converting Enzyme (ACE) assay for quantitative determination of ACE activity in human serum and plasma have been evaluated on the Abbott Alinity c system and ARCHITECT c16000 System. ACE (dipeptidyl carboxypeptidase) is a glycoprotein peptidyldipeptide hydrolase that cleaves histidylleucine dipeptide from angiotensin I, a relatively inactive decapeptide. The latter is converted to the potent vasoconstrictor, angiotensin II. ACE also inactivates bradykinin. Elevated levels of ACE activity occur in serum of patients with active sarcoidosis, and occasionally in premature infants with respiratory distress syndrome, in adults with tuberculosis, Gaucher’s disease, leprosy, and in many other pathologic conditions involving lung and liver diseases. Methods ACE hydrolyses urylacryloylphenylalanine-glycylglycine (FAPGG) to furylacryloylphenylalanine (FAP) and glycylglycine. Hydrolysis of FAPGG results in a decrease in absorbance at 340 nm. The ACE activity in the sample is determined by comparing the sample reaction rate to that obtained with the Sentinel ACE calibrator. Performances evaluation have been done following the latest CLSI guidelines protocols available. Results Summary of the analytical performances for the tested assay is reported in table below. Conclusions Analytical performances of Sentinel Diagnostics’ ACE assay demonstrate that quantitative determination of ACE activity in human serum and plasma are suitable for the routine measurement of the analyte on Abbott Alinity c system and ARCHITECT c16000 System.","PeriodicalId":10690,"journal":{"name":"Clinical chemistry","volume":"53 1","pages":""},"PeriodicalIF":9.3,"publicationDate":"2024-10-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142368788","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"B-016 Evaluation of pyridoxamine phosphate (PAMP) for AST/ALT reagents with recombinant human apoenzymes","authors":"T Oba, M Miyachi, Y Taketani, M Mizuguchi","doi":"10.1093/clinchem/hvae106.380","DOIUrl":"https://doi.org/10.1093/clinchem/hvae106.380","url":null,"abstract":"Background ApoAST/ALT, which have lost a coenzyme such as pyridoxal phosphate (PALP), are normally present at low levels in serum. To form the holoenzyme, IFCC recommends adding PALP into AST/ALT reagents. However, commercial reagents often do not use PALP due to its instability. In recent years, pyridoxamine phosphate (PAMP), a more stable coenzyme, has been marketed. However, the differences in holo formation ability and reagent stability between their coenzymes are not well understood. In this study, we focused on the preparation of PAMP and recombinant human ApoAST/ALT (rhApoAST/ALT), followed by fundamental investigations into their use as AST/ALT reagents. Methods PAMP was synthesized from pyridoxamine through an enzymatic reaction using pyridoxal kinase. The reaction product was purified to achieve a purity of >99% in HPLC. rhApoAST/ALT were expressed in Escherichia coli and purified with chromatography. AST/ALT assay was performed on a Hitachi 7180 automatic analyzer. PALP reagents for AST/ALT were designed to correspond with the final concentration of the IFCC method. PAMP reagents were prepared with PAMP instead of PALP. To evaluate the holo formation ability of PALP and PAMP reagents, we measured a dilution series of rhApoAST/ALT. To assess stability, the reagents were stored at 11°C for 3 months and then measured same. The reagents were subjected to content analysis by HPLC. Results Similar results were shown between coenzymes in the evaluation of holo formation ability, and high correlations were observed. As for the stability test, the values of PALP reagents decreased after 3 months, while the values of PAMP reagents did not. Content analysis showed that a spontaneous transamination occurred in PALP reagents. These results were observed at both AST and ALT reagents. Conclusions In AST/ALT reagents, both PAMP and PALP can activate apoenzymes. While PAMP is stable, PALP causes a decrease in reagent performance due to a spontaneous transamination.","PeriodicalId":10690,"journal":{"name":"Clinical chemistry","volume":"56 1","pages":""},"PeriodicalIF":9.3,"publicationDate":"2024-10-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142362939","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"B-194 Validation of a Diagnostic Test for the p.R337H Variant in the TP53 Gene by Nanopore Sequencing: Development of a Simplified Analysis Pipeline","authors":"M Andrade, A Belmok, L Silva, A Vieira, C Sousa, L Velasco, R Jácomo","doi":"10.1093/clinchem/hvae106.554","DOIUrl":"https://doi.org/10.1093/clinchem/hvae106.554","url":null,"abstract":"Background The genetic variant p.R337H in the TP53 gene is a pathogenic variant associated with Li-Fraumeni Syndrome and an increased risk of developing various types of cancer, including breast cancer. This variant is present in about 1 in every 300 individuals in the Southern region of Brazil, likely due to a founder effect. Amplicon sequencing via the Sanger methodology remains prevalent for the genotyping of such variants. The spectrum of applications for methods aimed at identifying human genetic variants spans from the exploration of pathogenic mutations to pharmacogenetic studies. Nevertheless, the use of conventional NGS techniques, such as Illumina, for this application is still financially costly. Third-generation sequencing by nanopores has low implementation costs and has seen expanded use recently, particularly due to genomic surveillance studies conducted during the pandemic, and after the release of the Q20+ chemistry, it may be an interesting alternative for human variant research. Thus, the present study aims to validate a diagnostic test for researching this variant through nanopore sequencing, including the development of a simplified analysis pipeline. Methods 24 blood samples with previously known results underwent nucleic acid extraction using the Maxwell extractor (Promega) with the Maxwell RSC Blood DNA kit. The extraction eluate was used in a PCR reaction. The exon 10 region of the TP53 gene was amplified using specific primers. Amplification confirmation was performed by agarose gel electrophoresis. Each sample received a barcode through a ligation reaction using NEB Blunt/TA Ligase Master Mix (NEB) according to the manufacturer's instructions. The sequencing library was prepared using the SQK-NBD114.24 kit (Nanopore) and sequenced on a flongle R10.1.4 using the Flongle Sequencing Expansion kit (EXP-FSE002). Sequencing was configured for 4 hours in the MinION control software (MinKnow). Basecalling was performed using Guppy_barcoder in simplex and duplex modes using the super accuracy algorithm. Reads were demultiplexed using a pipeline adapted from RAMPART and mapped to the reference using the minimap2 assembler implemented in Geneious. A mapping and variant calling workflow was created in Geneious software for batch analysis. Results The 24 PCR reactions produced amplicons of the expected size. The Rampart tool was adapted for exon 10 of the TP53 gene and allowed real-time monitoring of coverage per sample. Approximately 90 thousand reads were generated in total, with an average of 3600 reads per sample. The analyses showed that simplex and duplex basecalling methods are effective for analyzing the presence of the variant in question, however, simplex generated greater coverage. Comparison with Sanger sequencing revealed an accuracy of 100%. Conclusions Our results show that genotyping of the p.R337H variant in the TP53 gene can be performed using nanopore sequencing with the new Q20+ chemistry. The coverage obtained f","PeriodicalId":10690,"journal":{"name":"Clinical chemistry","volume":"33 1","pages":""},"PeriodicalIF":9.3,"publicationDate":"2024-10-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142369176","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"B-015 Analytical Verification of MR-proADM Biomarker in Human EDTA Plasma on Immunoassay Analyzer LIAISON® XL of Diasorin","authors":"R Ferrer-Costa, S Garriga-Edo Saraia, L Castillo Ribelles, T Balboa Lagunero, J C Ruiz-Rodríguez","doi":"10.1093/clinchem/hvae106.379","DOIUrl":"https://doi.org/10.1093/clinchem/hvae106.379","url":null,"abstract":"Background Adrenomedullin (ADM) is a potent vasodilator peptide of 52-aminoacid belonging to the calcitonin family of peptides. Midregional pro-ADM (MR-ProADM) is a precursor for ADM that circulates is in a 1:1 ratio with it, and proportionally represents its levels and activity. Increased ADM circulating concentrations have been described for several disease states, including dysfunction of the cardiovascular system and sepsis. However, reliable ADM quantification has been hampered by the short half-life, the existence of a binding protein, and its physical properties. Hence, quantificacion of MR-proADM represents a more suitable option. The biomarker MR-proADM has been shown to be an accurate prognostic marker for outcome prediction in patients with lower respiratory infections, sepsis, urinary tract infections, and kidney disease. The objective of this study was the verification of the metrological characteristics and the verification of the linearity of quantitative measurement range, as declared by the manufacturer, of the LIAISON® Brahms MR-proADMTM on Liaison® XL (DiaSorin). Methods The verification was performed with two levels of quality controls (QC) from the manufacturer (x̄L=1,19 nmol/L and x̄H=5,48 nmol/L), and residual EDTA plasma samples from patients with an active sepsis code. The evaluation included the assessment of: • Detection capability (including limit of quantification (LoQ) and limit of detection (LoD), measuring three samples at the manufacturer's stated concentration (0.21 nmol/L and 0.09 nmol/L, respectively) in duplicate within 4 runs). • Precision (CV, %) and accuracy (bias, %) within, measuring 10 replicates of each QC level; and between run, measuring 5 replicates for each QC level within 5 runs. • Linearity of the quantitative range of measurement (six intermediate samples prepared by mixing a high concentration sample- XH=9.64 nmol/L- and a low concentration sample, XL=0.59 nmol/L; measured in triplicate). All studies were performed in accordance with the recommendations of the Clinical Laboratory Standards Institute guidelines. Statistical analysis was performed using Microsoft Excel®, 2016. Results All results with diluted samples presented a signal (RLU) higher than that detected in samples without analyte, so the LoD was verified. The precision at the LoQ concentration was 26.7% (acceptance criteria was set as <20%). CV for within-run precision were 4.3% and 2.0% and for between-run precision 6.8% and 7.9% for low (L1) and high (L2) control level, respectively. The average bias was 3.60% for L1 and 4.23% for L2, meeting acceptance criteria (<8%). Linearity was confirmed between 0.59 and 9.64 nmol/L (R2 = 0.995). The biases between the expected and the obtained value were ≤7% through the quantitative range of measurement, except for the lowest concentration sample (close to the quantitation limit). Conclusions Our study showed good compliance with the metrological characteristics declared by the ","PeriodicalId":10690,"journal":{"name":"Clinical chemistry","volume":"2 1","pages":""},"PeriodicalIF":9.3,"publicationDate":"2024-10-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142362937","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"B-014 Performance evaluation of the automated cell image analyzer DI-60 for leukocyte differential count","authors":"S Chang, E Kim, Y Kim, D Won","doi":"10.1093/clinchem/hvae106.378","DOIUrl":"https://doi.org/10.1093/clinchem/hvae106.378","url":null,"abstract":"Background The Sysmex DI-60 system (Sysmex, kobe, Japan) is an automated cell image analyzer that captures cell images and analyzes them. The aim of this study was to evaluate the performance of DI-60 for leukocyte differential count in comparison with manual differential count or XN-20. Methods A total of 205 samples were analyzed. The agreement between DI-60 pre-classification and post-verification by medical technicians was determined. The correlation and ability to identify clinically important abnormal cells of DI-60 post-verification were evaluated. Results The overall agreement of DI-60 pre-classification was 84.7% (Table 1). The correlation between DI-60 post-verification and manual differential counts were exellent (r2 > 0.85) for neutrophil, lymphocyte, monocyte and eosinophil, but was very low for basophil (r2 = 0.3759). The correlation between DI-60 post-verification and leukocyte differential counts by XN-20 were exellent (r2 > 0.90) for neutrophil, lymphocyte and eosinophil. However, monocyte and basophil showed a relatively low correlation of r2 = 0.6343 and r2 = 0.3118, respectively but was very low for basophil (r2 0.3759). The ability of DI-60 post-verification to identify clinically important abnormal cells including blast, promyelocyte, myelocyte, metamyelocyte and NRBC demonstrated excellent efficiency (80.8 to 96.3%) except for NRBC (51.0%). Overall sensitivity was 63.7-88.2%, and specificity was 84.3-95.5% excluding NRBC (43.7%). Conclusions DI-60 showed excellent pre-classification and generally high correlation compared to manual leukocyte differential counts. Although additional verification with PBS review by experienced medical technician may still be required, the performance DI-60 makes it an efficient screening tool in clinical laboratories.","PeriodicalId":10690,"journal":{"name":"Clinical chemistry","volume":"14 1","pages":""},"PeriodicalIF":9.3,"publicationDate":"2024-10-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142362941","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"A-368 Feasibility of a Five-Minute Sample-to-Answer Basic Metabolic Panel for Point-of-Care Testing on the Fluxergy Analyzer","authors":"E Charrault, X Zhen, E Dumitrescu, P Prakash, E Pineda, D Caron, B Hernandez, P Lamberg, K Sathyamoorthy, S Gandhi, A Singh, T Patel, R Heltsley","doi":"10.1093/clinchem/hvae106.362","DOIUrl":"https://doi.org/10.1093/clinchem/hvae106.362","url":null,"abstract":"Background Adoption of point-of-care (POC) platforms has been limited by their high cost and limited menu (e.g., molecular only). Fluxergy has developed a cost-effective multimodal POC platform capable of performing the most common clinical chemistry, immunochemistry, hematology, molecular tests in CLIA-waived settings. This method comparison study describes the performance of the Fluxergy’s feasibility stage basic metabolic panel (BMP) with contrived samples compared to the Siemen’s epoc® Blood Analysis System. Methods Fluxergy Analyzer: The Fluxergy Analyzer is a rapid and cost-effective multimodality platform that can perform lab-quality PCR, clinical chemistry, immunochemistry, and cytometry in a single instrument and if required, in a single cartridge. Fluxergy BMP: Fluxergy BMP rapidly quantifies key electrolytes (K+, Na+, Ca2+, Cl-) and metabolites (glucose, creatinine, and BUN) from whole blood in under five minutes on the Fluxergy Analyzer, utilizing the Fluxergy test cards with screen-printed electrochemical sensor. Feasibility Study: A method comparison study was conducted to assess the performance of the feasibility stage Fluxergy BMP using contrived samples generated by spiking known concentrations of K+, Na+, Ca2+, Cl-, and glucose into buffer solutions. Samples were tested in triplicate with both the Fluxergy BMP and Siemen’s epoc® Blood Analysis System at three different concentrations across the reportable range of the epoc® Blood Analysis System. Results The feasibility stage Fluxergy BMP demonstrated strong correlation across the reportable range for K+, Na+, Ca2+, Cl-, and glucose versus the epoc® Blood Analysis System (Table 1). The error percentage for all samples tested except one glucose sample (9.5 mM) was less than 10%. The coefficient of variance for all samples tested except one Ca2+ sample (0.44 mM) was less than ten percent. Conclusions This feasibility data demonstrates the Fluxergy Analyzer is well-suited to perform rapid and accurate metabolic testing from whole blood samples at the POC.","PeriodicalId":10690,"journal":{"name":"Clinical chemistry","volume":"23 1","pages":""},"PeriodicalIF":9.3,"publicationDate":"2024-10-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142362966","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"A-171 Novel Fluorometric Assay for Pediatric Anti-Factor Xa Testing: Minimizing Bilirubin and Hemolysis Interference in Whole Blood","authors":"V Pamula, M Basmajian, A Ullal, L Nichols, S Adhikari, M Moser, S Emani, S Emani","doi":"10.1093/clinchem/hvae106.169","DOIUrl":"https://doi.org/10.1093/clinchem/hvae106.169","url":null,"abstract":"Background High bilirubin levels are common in pediatric patients and are noted in patients undergoing extracorporeal membrane oxygenation (ECMO). Acute anticoagulation therapy necessitates frequent monitoring of anti-Factor Xa activity (aFXa), and high bilirubin levels cause interference in chromogenic assays. Moreover, large volumes of blood lead to iatrogenic anemia in pediatric and neonatal patients. We developed a fluorescence assay to measure aFXa activity on a near-patient digital microfluidic (DMF) platform using low volume whole blood samples (< 50 μL) and present the results of interference of bilirubin and hemolysis on aFXa activity. Methods aFXa assay was performed on a DMF cartridge wherein 50 μL whole blood was loaded into the cartridge which separates plasma droplets through agglutination within the cartridge followed by incubation with a droplet containing exogenous FXa and a fluorogenic substrate. The fluorescence is inversely proportional to the concentration of heparin in the sample. All required reagents for the aFXa assay are dried within the cartridge, allowing for integrated sample preparation and assay automation for point of care use. To test the interference of bilirubin, we spiked into whole blood different concentrations of unconjugated bilirubin, ranging from 0-40 mg/dL. We also tested hemoglobin interference by testing 4 different concentrations of hemolysate (0-1,000 mg/dL) on the aFXa assay. All experiments were performed with 6 replicates of each concentration. Results Across each bilirubin concentration, we saw uniform measurements of aFXa activity (see Table). We observed < 5% CV in aFXa activity with 0, 13.33 and 26.67 mg/dL bilirubin levels. All measured unfractionated heparin values for hemolysate interference were <5% CV. Conclusions Our interference results suggest that high bilirubin and hemolysate levels do not interfere with our novel aFXa fluorescence assay. Further clinical studies are underway to establish clinical performance.","PeriodicalId":10690,"journal":{"name":"Clinical chemistry","volume":"14 1","pages":""},"PeriodicalIF":9.3,"publicationDate":"2024-10-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142369092","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"A-257 Revolutionizing Fungal Infection Diagnosis: A Sensitive, Cost-Effective, and Time-Efficient Solution","authors":"K Ramakrishnan, M Maung, V Ezike, P Poudel, R Senthilvelan, C Cui, J Mccabe, S Sackeyfio, A Senejani, E Kirkor, S Sinha","doi":"10.1093/clinchem/hvae106.254","DOIUrl":"https://doi.org/10.1093/clinchem/hvae106.254","url":null,"abstract":"Background Fungal infection detection is essential for optimizing therapeutic strategies, preventing complications, enhancing overall health, and alleviating the financial burden, and preventing transmission. According to the CDC, fungal illness resulted in 75,000 hospital admissions and 9 million outpatient visits annually, with 7,199 deaths predicted in the United States in 2021. The rising prevalence of fungal infections, influenced by factors such as population expansion and evolving treatment strategies poses a growing challenge to achieving an early and accurate diagnosis. There is the crucial need for improved detection methods to promptly identify various fungal infections, including Candida species, spanning from skin issues to potentially fatal systemic diseases, ultimately reducing associated morbidity and mortality. Severe and profound infections with life-threatening potential are common in individuals with compromised immune systems and those who are hospitalized. This risk is notably elevated among patients with organ transplants, immunosuppressive treatments, diabetes, recent broad-spectrum antibiotic usage, catheter utilization, and extended hospitalization periods. Our advancement seeks to introduce innovative methods leveraging carbon-based nanomaterials for the detection of fungal infections, offering distinct advantages including cost-effectiveness, ease of operation, and time efficiency compared to conventional techniques. Methods Our technology utilizes carbon-based nano materials, a promising advancement in nanotechnology to detect fungal infections, especially Candida manifestations. The samples are brought into contact with pre-probed carbon-based nano materials sensors maintained at a specific annealing temperature, facilitating nucleic acid hybridization. Subsequently, alterations in the carbon nanotube's electrical signal are observed, indicating a change in conductivity. This change can allow for the measurement of a voltage difference across a continuous flow of electricity, correlating with the concentration of targets present in the sample, thereby enabling the detection of specific microorganisms. No change was noticed in electrical properties of carbon-based nano material when negative experiments were conducted. Results The initial findings showed the presence of specific microorganisms within the sample in less than 15 minutes with carbon-based nano materials sensors. Comparative analysis of the electrical signal magnitude values obtained from carbon nanotube measurements against the corresponding CT values from quantitative PCR (qPCR) provides insights into the correlation between the electrical readouts and molecular quantification, offering a comprehensive evaluation of the nanotube-based detection system's performance in diagnostics procedure. Conclusions Unlike conventional blood culture and qPCR, our technology, delivers outcomes with heightened sensitivity, cost-effectiveness, and time efficiency. Our t","PeriodicalId":10690,"journal":{"name":"Clinical chemistry","volume":"25 1","pages":""},"PeriodicalIF":9.3,"publicationDate":"2024-10-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142369122","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"A-064 Improved Insulin production in Advanced glycation end-products (AGEs)-induced RIN-m5F pancreatic β-cells by using a Flavonoid-Rich Medicinal Plant, Kaempferia parviflora treatment","authors":"K Chumroonsiri, A Chiabchalard","doi":"10.1093/clinchem/hvae106.063","DOIUrl":"https://doi.org/10.1093/clinchem/hvae106.063","url":null,"abstract":"Background Advanced glycation end-products (AGEs) are glycated proteins that react with reducing sugar which causes the damages of function and cellular signaling of insulin production in insulinoma cells. AGEs also contribute to hyperglycemia-induced insulin resistance in type 2 diabetes. AGEs gradually form during the chronic high blood glucose condition, which is a major risk factor and harmful to β-cells function and survival, and could alter the insulin signaling and downregulate the insulin-responsive transcription factors including PDX-1, Glut-2, GCK, and Pre-INS. Kaempferia parviflora, also known as black ginger and flavonoid-rich medicinal plant, has beneficial properties for type 2 diabetes patients as a counteracts the detrimental effects of AGEs and improves the insulin production in pancreatic β-cells. Thus, we hypothesize that treated β-cells with K. parviflora, which is the most effective flavonoid-containing plant might protect against the damage of AGEs-induced insulin impairment, and these effects are also responsible for improved insulin synthesis and the essential transcription factors in pancreatic β-cells. Methods Rat pancreatic β-cell line RIN-m5F (ATCC CRL-2058) were maintained and cultured in RPMI 1640 medium containing 10% (v/v) FBS, 1% penicillin-streptomycin at 37◦C in a humidified atmosphere containing 5% CO2. RIN-m5F cells were pre-treated with the K. parviflora at concentrations 0, 25, and 50 µg/ml for 48 hours, and afterward, exposure to 250 µg/ml AGEs treatment. To quantify the bioactive compound of K. parviflora, total phenolic and flavonoid content assays were performed and the transcriptional changes of insulin-related genes (PDX-1, Glut-2, GCK, and Pre-INS) were determined by using real-time qRT-PCR. Results K. parviflora has shown a high phenolic content and flavonoid content was 50.43 ± 1.74, 26.66 ± 1.18 mg GAE/g Dry weight of sample, respectively. For the transcriptional changes of insulin-related genes, the levels of gene expression in the regulators of insulin secreting pathway (PDX-1, Glut-2, GCK, and Pre-INS) were significantly downregulated after exposure to AGEs-treated, while the pre-treatment of K. parviflora was showed statistically difference. PDX-1, Glut-2, GCK, and Pre-INS expressions were significantly upregulated after pre-treated with 50 µg/ml K. parviflora. However, under the AGEs-treated alone condition, the transcriptional levels were significantly decreased as compared to the control (untreated). Taken together, these results suggest K. parviflora could be a potential natural medicine to ameliorate the damage of insulin production in AGEs-induced β-cell damage and highlight promising alternatives for therapeutic avenue against diabetes. Conclusions Treated RIN-m5F pancreatic β-cells with K. parviflora, a flavonoid-rich medicinal plant, could upregulate the insulin-responsive transcription factors and improve the insulin production in damaging AGEs-induced pancreatic β-cells. Our findings","PeriodicalId":10690,"journal":{"name":"Clinical chemistry","volume":"77 1","pages":""},"PeriodicalIF":9.3,"publicationDate":"2024-10-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142368790","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}