Chemiluminescent Enzyme Immunoassay Method Research Articles

Diagnostic Performance of CLEIA Versus FEIA for KL-6 Peripheral and Alveolar Concentrations in Fibrotic Interstitial Lung Diseases: A Multicentre Study.

Interstitial lung diseases (ILD) is a group of lung disorders characterized by interstitial lung thickening due to inflammatory and fibrotic processes. Krebs von den Lungen-6 (KL-6) is a molecule secreted by damaged type II alveolar pneumocytes in the alveolar space. The goal of the present study was to compare two detection methods of KL-6 in both bronchoalveolar lavage (BAL) and serum from ILD patients at the moment of diagnosis. Patients with suspicious of ILD and followed at two Italian referral centres for rare lung diseases were included in the study. BAL fluid and serum were collected and analysed by chemiluminescent enzyme immunoassay (CLEIA) and fluorescent enzyme immunoassay (FEIA) methods provided by Tosoh Biosciences. A total of 158 (mean age ± standard deviation, 61.5 ± 13.7, 65 females) patients were enrolled. A total of, 36 had diagnosis of idiopathic pulmonary fibrosis (IPF), 74 sarcoidosis, 15 connective tissue disease-ILD (CTD-ILD) and 33 other ILD. Diagnostic agreement between two methods was demonstrated for both BAL (r = 0.707, p < 0.0001) and serum (r = 0.816, p < 0.0001). BAL KL-6 values were lower than serum (p < 0.0001). IPF patients had higher serum KL-6 concentration than other ILDs (p = 0.0294), while BAL KL-6 values were lower in IPF than in non-IPF (p = 0.0023). This study explored KL-6 concentrations through the CLEIA method in serum and BAL of patients with various ILDs, showing significant differences of biomarkers concentrations between IPF and other non-IPF ILDs. Our findings are promising as they provided further knowledge concerning KL-6 expression across different ILDs and may suggest its utility in differential diagnosis.

Investigating the cut-off values of captopril challenge test for primary aldosteronism using the novel chemiluminescent enzyme immunoassay method: a retrospective cohort study

The measurement evolution enabled more accurate evaluation of aldosterone production in hypertensive patients. However, the cut-off values for novel assays have been not sufficiently validated. The present study was undertaken to validate the novel chemiluminescent enzyme immunoassay for aldosterone in conjunction with other methods. Moreover, we also aimed to establish a new cut-off value for primary aldosteronism in the captopril challenge test using the novel assay. First, we collected 390 plasma samples, in which aldosterone levels measured using liquid chromatography-mass spectrometry ranged between 0.18 and 1346 ng/dL. The novel chemiluminescent enzyme immunoassay showed identical correlation of plasma aldosterone with liquid chromatography-mass spectrometry, in contrast to conventional radioimmunoassay. Further, we enrolled 299 and 39 patients with primary aldosteronism and essential hypertension, respectively. Plasma aldosterone concentrations measured using the novel assay were lower than those measured by radioimmunoassay, which resulted in decreased aldosterone-to-renin ratios. Subsequently, positive results of the captopril challenge test based on radioimmunoassay turned into “negative” based on the novel assay in 45% patients with primary aldosteronism, using the conventional cut-off value (aldosterone-to-renin activity ratio > 20 ng/dL per ng/mL/h). Receiver operating characteristic curve analysis demonstrated that aldosterone-to-renin activity ratios > 8.2 ng/dL per ng/mL/h in the novel assay was compatible with the conventional diagnosis (sensitivity, 0.874; specificity, 0.980). Our study indicates the great measurement accuracy of the novel chemiluminescent enzyme immunoassay for aldosterone, and the importance of measurement-adjusted cut-offs in the diagnosis of primary aldosteronism.

Analytical Performance of a Novel Latex Turbidimetric Immunoassay, "Nanopia TARC", for TARC/CCL17 Measurement: A Retrospective Observational Study.

Thymus- and activation-regulated chemokine (TARC, also known as CCL17) is used as a biomarker for atopic dermatitis. The methods currently used for its measurement are complex, time-consuming, and require large machinery, warranting the need for a method that is simple, has a quick turnaround time, and requires less complex machinery. We evaluated the analytical performance of a novel latex turbidimetric immunoassay method, "Nanopia TARC", on 174 residual serum samples from patients with skin or allergic diseases. This evaluation included the assessment of the limit of blank/detection/quantification (LOB/D/Q), precision, accuracy, linearity, interference, and commutability between Nanopia TARC and "HISCL TARC", based on the chemiluminescent enzyme immunoassay (CLEIA) method. The LOB/D/Q values were 13, 57, and 141 pg/mL, respectively. The coefficient of variation of the repeatability was 0.9-3.8%, and that of the intermediate precision was 2.1-5.4%. The total error of the accuracy was 1.9-13.4%. The linearity was 141 and 19,804 pg/mL for TARC. The correlation coefficient between Nanopia TARC and HISCL TARC determined using the Passing-Bablok regression analysis was 0.999. Furthermore, the concordance of diagnostic criteria with AD was 92%. Nanopia TARC was confirmed to have the same analytical performance for TARC measurement as the existing CLEIA method.

JS-HR-9: A NEW ERA OF PROGRESS IN PRIMARY ALDOSTERONISM TREATMENT, A NEW ALDOSTERONE ASSAY, AND A CLINICAL PRACTICE GUIDELINE

Primary aldosteronism (PA), one of the mineralocorticoid receptor (MR)-associated hypertension, is a highly prevalent secondary hypertension that accounts for 5–15% of all hypertension and is a severe hypertension that is prone to cerebrovascular and cardiovascular disease. Therefore, it is expected that the pathophysiology and treatment strategies for PA will be elucidated. In recent years, the major changes in PA have been in treatment and diagnosis in Japan. Bilateral or non-surgical unilateral PA is treated with mineralocorticoid receptor antagonist (MRA). Esaxerenone, a non-steroidal MRA, is now widely used in Japan for hypertension including PA. It has been reported that add-on treatment using esaxerenone with maximal tolerable doses of a renin-angiotensin system inhibitor reduced the urinary albumin creatinine ratio in patients with type 2 diabetes mellitus (ESAX-DN), suggesting a renoprotective effect against diabetic nephropathy. Several clinical studies of another non-steroidal MRA, finerenone(FIDELIO-DKD, FIGARO-DKD, and FIDELITY), have shown its renoprotective effect against diabetic nephropathy as well as cardiovascular events, particularly hospitalization for heart failure in patients with type 2 diabetes and chronic kidney disease. Although there are no reports of clinical studies on finerenone for PA, finerenone may be used in the future for type 2 diabetes patients with PA, as obesity, glucose intolerance, and sleep apnea are common complications in patients with PA. Secondly, radiofrequency ablation of macroscopic adrenal tumors has been reported as an alternative treatment for PA to lower blood pressure and plasma aldosterone levels. This treatment has been covered by health insurance providers in Japan since April 2022, but long-term outcomes need to be validated. In diagnosis, serum and urine aldosterone has been measured in Japan since early 2021 by the two-step sandwich chemiluminescent enzyme immunoassay (CLEIA) method. This new CLEIA method correlates well with LC-MS/MS values and is expected to be more accurate. As CLEIA serum aldosterone levels are lower than those measured with a previous radioimmunoassay, the Japan Endocrine Society issued the 2021 Primary Aldosteronism Clinical Guidelines, which included a provisional positive value for the diagnosis of PA. More than ever, the diagnosis of PA needs to be carefully considered. This CLEIA method is currently available only in Japan. This presentation will cover the latest treatments, diagnostic procedures, and other hot topics in PA as described above.

PS-C21-10: A LC-MS/MS-EQUIVALENT VALUE OF ALDOSTERONE MEASUREMENT BY A NEW CHEMILUMINESCENT ENZYME IMMUNOASSAY USING A TWO-STEP SANDWICH METHOD

Primary aldosteronism (PA) is the most frequent cause of secondary hypertension. Patients with PA have an increased risk of cardiovascular events, such as cerebral infarction, myocardial infarction, and chronic kidney disease, compared to those with essential hypertension, and early diagnosis makes PA a treatable disease. In clinical practice, accurate measurement of plasma aldosterone concentration (PAC) is crucial because entire diagnostic tests critically depend on aldosterone concentrations. Recently, we developed a new chemiluminescent enzyme immunoassay (CLEIA) using a two step sandwich method to measure aldosterone concentrations. We measured PAC in blood samples from patients using a radioimmunoassay (RIA) and the CLEIA (with current and newly improved reagents) as well as liquid chromatography tandem mass spectrometry (LC MS/MS). Based on the results of the Passing Bablok regression analysis, the aldosterone levels measured using CLEIA with the new reagents and those measured by LC MS/MS were found to be significantly correlated. However, aldosterone levels were lower with the improved CLEIA method than with RIA and CLEIA using the current reagent. Since April 2021, the plasma aldosterone concentration has been measured by CLEIA in Japan. As a provisional response due to changes of the measurement method, the Japan Endocrine Society Clinical Practice Guideline for the Diagnosis and Management of Primary Aldosteronism 2021 was announced. This time, we will report on the new measuring method.

An autopsy case of fatal insulin preparation overdose

Insulin preparations, a group of drugs used to treat diabetes, are of great forensic interest because they are difficult to detect in blood while they are sometimes intentionally administered in large doses for suicide or murder. In this study, we validated a quantification method for insulin glargine and insulin lispro using liquid chromatography-tandem mass spectrometry (LC-MS/MS) in the blood sample from a case of suicidal overdose with insulin. Case: A woman in her 30s had a history of mental illness and diabetes. A day before her death, she informed her partner that she had self-administered large doses of insulin preparations and prescription drugs. Bovine insulin was used as the internal standard (IS). The blood sample (250 μL) was added to the diluent and IS solution, followed by 250 μL each of hexane and N,N-dimethylformamide, and the mixture was centrifuged (10000 g × 10 min). The supernatant was collected and 900 μL of 5% ammonium solution was added. The extract was analyzed using Nexera 30A, LCMS8060 (Shimadzu), L-column3 ODS C18 metal-free column (150 × 2.1 mm, 3 μm), ionization method: electrospray ionization+, multiple reaction monitoring conditions: insulin glargine [M + 6H]6+ 1011.5 > 1179.4, insulin lispro[M + 6H]6+ 968.9 > 217.0, bovine insulin (IS) [M + 6H]6+ 956.3 > 1120.9. The autopsy revealed no evidence of fatal injuries. Coronary artery stenosis was observed as a finding of endogenous disease, but there were no significant histopathological changes in the myocardium. Biochemical tests revealed the following: blood glucose level, 6 mg/dL; glucose in the vitreous region, 6 mg/dL; glycated hemoglobin (HbA1c), 11.2%; blood insulin level estimated using the chemiluminescent enzyme immunoassay method, 1.03 μU/mL (6.27 pmol/mL); and C-peptide concentration, 0.6 ng/mL (16.8 pmol/L). A drug screening test and simple quantification using LC-MS/MS detected many prescription drugs, but none of them were at intoxicating concentrations. The insulin analysis showed insulin glargine concentrations of 429 μU/mL in femoral blood and 1362 μU/mL in cardiac blood, and insulin lispro was detected in femoral and cardiac blood below the lower limit of quantification (<50 μU/mL). The cardiac/peripheral blood ratio, a measure of postmortem redistribution, was 3.2. Although the therapeutic ranges of insulin glargine and insulin lispro have not been defined, the femoral and cardiac blood concentrations of insulin lispro in this case was lower than the serum concentration (155 μU/mL) listed in the package insert. However, while the femoral and cardiac blood insulin glargine concentrations were considerably higher than the serum concentration (15 μU/mL) listed in the package insert. Considering that insulin is easily degraded in whole blood due to hemolysis, it is possible that insulin concentrations were even higher at the time of administration and death than those observed in autopsy specimens on the third postmortem day. However, since insulin lispro is a fast-acting analog of insulin (1–3 hours), it may have contributed to hypoglycemia but was not the direct cause of death, and hypoglycemia due to insulin glargine overdose was the most likely cause of death in this case. We quantified insulin lispro and insulin glargine in a case of suicide to determine insulin glargine's involvement in causing death. In future, it is necessary to examine postmortem degradation and redistribution of insulin and consider the appropriate concentration in postmortem samples of insulin poisoning.

Antibody Sustainability in SARS-CoV-2 Healthcare Professionals' Patient Cohort

In this study, it was aimed to evaluate one-year follow-up of severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) specific antibodies formed against the virus binding site, in a coronavirus disease-2019 (COVID-19) positive case cohort (n= 413) between the period March 2020 to December 2020 in Manisa Celal Bayar University Hospital, until July 2021. SARS-CoV-2 antibodies were determined by the chemiluminescent enzyme immunoassay (CLIA) method. Values of 1.0 and above were considered positive. Chi-square tests and Joinpoint regression analysis (version 4.7.0) were used in the statistical analyses. The mean age of the participants was 34.9 ± 9.3 and 60.2% of them were women. Between 21-30 days after the diagnosis of COVID-19, total antibody level was above the threshold value in 72.2% (n= 126) of the participants, while this rate increased to 79.1% (n= 240) in 31-60 day interval. In the following period, this rate decreased to 38.8% (n= 108) in days 211st to 240th. Antibody response could not be detected in 76 (20.7%) of 367 employees who have initially been followed up. The percentage of total antibody positivity prevalence ranged from 98.9% to 96.1% in the 31-210th day after diagnosis, in the follow-up of 291 employees whose total antibody positivity was detected after diagnosis. According to the results of the Joinpoint regression analysis, after the diagnosis of COVID-19, the curve showing the percentage of antibody positivity was broken at two points: The first breaking point was observed in 181-210th days (6-7 months) (p= 0.069), and the second breaking point was in 271-300th days (9-10 months) (p< 0.001). As a result, the highest antibody positivity rates were detected after the 30th day of the disease onset and antibody positivity was maintained in the first seven months after diagnosis; the antibody positivity rate decreased to 25% at the end of the first year.

Türkiye’de bir üçüncü basamak hastanede sağlık çalışanlarında SARS-CoV-2 seroprevalansı

Objective: The aim of this study is to investigate the previous four months (March-July 2020) SARS-CoV-2 infection rate, seroprevalence and the variables affecting these in HCWs in a university hospital. Methods: The present study is a SARS-CoV-2 seroprevalence study on HCWs working in a tertiary hospital during the first stage (March-July 2020) of the outbreak in Turkey. The presence of IgM and IgG antibodies against the spike structure of the virus was investigated by the chemiluminescent enzyme immunoassay (CLIA) method using the commercial antibody kit (COV2T, Siemens®, Tarrytown, NY, US). Participants’ socio-demographic characteristics, health status, lifestyle, risky occupational and social and personal protective equipment (PPE) usage were independent variables of the study. Chi-square test and Fisher’s exact test were used in univariate analyzes, and accepted type 1 error value was 0.05. The analyzes were made using the SPSS 23.0 package program. Results: 1177 out of a total of 1702 health workers participated in the study. Participation rate was 69.1% . The mean age of the study group was 35.3 ± 9.8 and 62.7% were females. SARS-CoV-2 infection rate detected by nucleic acid amplification test (NAAT-PCR) or antibody test (Elisa) was (18/1177) 1.5%; The seroprevalence of SARS-CoV-2 was 1.01%. 17% of the entire SARS-CoV-2 cases were asymptomatic. The highest infection prevalence was significantly higher in auxiliary health workers (3.7%) compared to other groups. The presence of symptoms HCW’s and their family members that did not exist before in the last 15 days, being overweight or obese and consulting as contacted person in survelliance unit were significantly related to having SARS-CoV-2 infection (p

Conformity of cleia and clia HBsAg quantitative assay with qPCR gold standard in hepatitis B patients

Quantitative HBsAg assay is an alternative marker of potential viremia and monitoring response to antiviral treatment. Various methods have been developed to assess HBsAg quantification. This cross-sectional observational research aims to understand the performance of quantitative HBsAg assay with CLEIA (Chemiluminescence Enzyme Immunoassay) and CLIA (Chemiluminescence Immunoassay) methods and their compliance with qPCR. Serum samples were obtained from 69 Hepatitis B patients at the Outpatient Clinic. Quantitative HBsAg assay was performed with Sysmex HISCL-5000 and Mindray CL-900i, HBV DNA by using GeneXpert® HBV Viral Load. The results showed that the HBsAg value of the CLEIA method ranged from 9.77 to &gt;2,500 IU/mL, while the CLIA method ranged from 19.97 to &gt;50,000 IU/mL. Analysis of the CLEIA and CLIA methods showed agreement ((LoA: -5014.01 – 1815.33) with the relationship between the two methods showed a positive correlation (p &lt;0.05). There is a positive correlation between HBsAg results using CLEIA method and HBV DNA (p &lt;0.05) and also between the HBsAg CLIA method and HBV DNA (p &lt;0.05).

Krebs von den Lungen-6 as Disease Severity Marker for COVID-19 Patients: Analytical Verification and Quality Assessment of the Tosoh AIA-360 Compared to Lumipulse G600II.

Background: Krebs von den Lungen-6 (KL-6) has been proposed as a disease severity marker of COVID-19. All research articles reported the KL-6 assay detected through Fujirebio reagents by Lumipulse G600/G1200 instrument. In the present study, KL-6 assay was analysed through Tosoh AIA-360 and compared with analytical results by Lumipulse G600 in a population of COVID-19 patients. Materials and methods: Sixty-four patients (median age, IQR 67 (58–76) years), all hospitalized for COVID-19 interstitial pneumonia at Siena COVID Unit. KL-6 was measured by two methods, chemiluminescence enzyme immunoassay (CLEIA) and fluorescent enzyme immunoassay (FEIA) method by Lumipulse G600 II and AIA 360 systems, respectively. Results: KL-6 concentrations evaluated by Lumipulse G600II were significantly higher in severe than those in non-severe patients (p < 0.0001) as well as evaluating by AIA360 (p < 0.0001). Receiver operating curve (ROC) curve analysis showed that KL-6 concentrations, by Lumipuse G600II, distinguished severe from non-severe COVID-19 patients with an area under the curve (AUC) of 99.8% and the best cut-off value was 448 U/mL. AUROC between severe and non-severe COVID-19 patients using T0 KL-6 concentrations by AIA360 was 97.4% and the best cut-off value was 398 U/mL. According to T0 KL-6 concentrations in COVID-19 patients, Bland–Altman difference analysis revealed a mean bias of 78 ± 174.8; while using T1 KL-6 concentrations in COVID-19 patients, Bland–Altman difference analysis revealed a mean bias of 48 ± 126 (95% limits of agreement −199–295) between the Lumipulse G600 II and the AIA360 systems. Conclusions: In conclusion, our study demonstrated that CLEIA and FEIA methods for serum KL-6 detection are comparable and reliable. KL-6 was confirmed as an easily detectable and effective biomarker to identify severe COVID-19 patients.

Antibody Responses to BNT162b2 Vaccination in Japan: Monitoring Vaccine Efficacy by Measuring IgG Antibodies against the Receptor-Binding Domain of SARS-CoV-2.

ABSTRACTTo fight severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), which causes coronavirus disease 2019 (COVID-19), mass vaccination has begun in many countries. To investigate the usefulness of a serological assay to predict vaccine efficacy, we analyzed the levels of IgG, IgM, and IgA against the receptor-binding domain (RBD) of SARS-CoV-2 in the sera from BNT162b2 vaccinated individuals in Japan. This study included 219 individuals who received two doses of BNT162b2. The levels of IgG, IgM, and IgA against RBD were measured by enzyme-linked immunosorbent assay before and after the first and second vaccination, respectively. The relationship between antibody levels and several factors, including age, gender, and hypertension were analyzed. Virus-neutralizing activity in sera was measured to determine the correlation with the levels of antibodies. A chemiluminescent enzyme immunoassay (CLEIA) method to measure IgG against RBD was developed and validated for the clinical setting. The levels of all antibody isotypes were increased after vaccination. Among them, RBD-IgG was dramatically increased after the second vaccination. The IgG levels in females were significantly higher than in males. There was a negative correlation between age and IgG levels in males. The IgG levels significantly correlated with the neutralizing activity. The CLEIA assay measuring IgG against RBD showed a reliable performance and a high correlation with neutralizing activity. Monitoring of IgG against RBD is a powerful tool to predict the efficacy of SARS-CoV-2 vaccination and provides useful information in considering a personalized vaccination strategy for COVID-19.IMPORTANCE Mass vaccination campaigns using mRNA vaccines against SARS-CoV-2 have begun in many countries. Serological assays to detect antibody production may be a useful tool to monitor the efficacy of SARS-CoV-2 vaccination in individuals. Here, we reported the induction of antibody isotype responses after the first and second dose of the BNT162b2 vaccine in a well-defined cohort of employees in Japan. We also reported that age, gender, and hypertension are associated with differences in antibody response after vaccination. This study not only provides valuable information with respect to antibody responses after BNT162b2 vaccination in the Japanese population but also the usefulness of serological assays for monitoring vaccine efficacy in clinical laboratories to determine a personalized vaccination strategy for COVID-19.

Хемилюминесценция функционализированной поверхности графена

To analyze the modification of the functionalized surface of graphene by protein molecules, a chemiluminescent enzyme immunoassay method was proposed. Using the example of functionalized graphene (FG) purification, the possibilities of chemiluminescent control of the state of its surface are shown. Methods for purifying FG from protein molecules with the restoration of the ability to resorb protein molecules are discussed. It has been shown that the FG surface can be cleaned from sorbed proteins, and a biosensor can be designed again based on such purified graphene, including with a different specificity. Therefore, the graphene sensor can be used repeatedly.

Stimulation of sCD14-ST (Presepsin) Secretion By Peripheral Blood Mononuclear Cells after Candida Albicans Lysate and Lipopolysaccharide Exposure

Background:Сluster of differentiation 14 (CD14) is a specific high-affinity receptor for lipopolysaccharides (LPSs) on monocytes, macrophages and granulocytes. Soluble CD14 subtype (sCD14-ST, Presepsin) is a shortened soluble N-terminal fragment of CD14. Levels of sCD14-ST have a strong correlation with severe bacterial infection which can be used as diagnostic tool. However, CD14 recognize not only LPS, but also a variety of ligands associated also with Gram-positive bacteria, fungi, and viruses. Here we report data about impact of C.albicans antigens on monocytes and presepsin level in vitro .Donors and Methods:Peripheral Blood Mononuclear Cells (PBMCs) from 19 healthy volunteers (12 women,7 men, age 20-38, median 28 years) were isolated on media gradient Fiqoll-Histopaque (p=1.077 g/ml) by standard protocol. Fraction of monocytes in PMBC was defined by light scatter properties on flow cytometry (BD FACS Canto II, USA). PBMCs with 0.5 x 106 of monocytes in RPMI-1640 (with 10% of autologous serum) were incubated in 5% CO2, 370C with LPS (GIEM, Moscow, Russia) at the final concentration - 1 µcg/ml (“LPS” tube); with Candida albicans lysates (Sanum-Kehlbeck GmbH & Co, Germany) at the final concentration 200 ng/ml (“ C.albicans ” tube) and control tube with RPMI-1640 (with 10% of autologous serum) and without PMBC (“Serum” tube). Twenty-four hours and 48 hours after stimulation a 150 µL-samples were collected and centrifuged. Presepsin concentration were measured using a chemiluminescent enzyme immunoassay method, performed on PATHFAST analyzer (Mitsubishi Chemical Medience Corporation, Tokyo, Japan). Mann-Whitney U test was used for nonparametric data analysis between two groups. Wilcoxon signed-rank test was used to compare repeated measures. A p-value less than 0.05 was considered as significant.Results:Median (with ranges) of sCD14-ST concentration 24 hours after beginning of stimulation was 59,8 pg/ml (29,7-140), 84 pg/ml (38,8-133) and 34,6 pg/ml (18,5-81,8) for “LPS”, “ C.albicans ” and “Serum” tube respectively. 48 hours after beginning of stimulation sCD14-ST concentration increased to 85,8 pg/ml (45-146), 114 pg/ml (62,3-198) and 40,8 pg/ml (25,3-74,8) also for “LPS”, “ C.albicans ” and “Serum” tube respectively. According to our data (see Figure 1), the level of sCD14-ST in supernatant after 24 and 48 hours in « C.albicans » tube was significantly higher than in «LPS» (p <0.05) and in serum tube (p<0.0001).Conclusion:Here we report data about impact of C.albicans antigens on monocytes and sCD14-ST level in vitro . Obtained data show us that not only bacterial infection can lead to increased level of sCD14-ST (Presepsin) and lead us to preposition that extra-high level of sCD14-ST (Presepsin) can be found in patients with severe fungal infections. [Display omitted] DisclosuresNo relevant conflicts of interest to declare.

PIVKA-II: A biomarker for diagnosing and monitoring patients with pancreatic adenocarcinoma

BackgroundPancreatic adenocarcinoma (PDAC) is an incurable cancer without adequate tumor markers. Our previous study has showed a better diagnostic performance of Protein Induced by Vitamin K Absence II (PIVKA-II) compared to currently used PDAC biomarkers. To corroborate our previous data with a larger sample size and to assess a possible role of PIVKA-II in predicting surgical success. Additionally, to further evaluate the hypothesis of a direct PIVKA-II production by PDAC cells, we examined PIVKA-II tissue expression in a case of PDAC using immunofluorescence.MethodsWe enrolled 76 newly diagnosed PDAC patients and selected 11 patients to determine PIVKA-II levels also after surgical resection. An immunofluorescence (IF) study of PIVKA-II tissue expression was carried out in one of them. PIVKA-II serum values were measured by chemiluminescent enzyme immunoassay method (CLEIA) on LUMIPULSE G1200 (Fujirebio-Europe, Belgium).ResultsPIVKA-II serum levels were above the cut-off at baseline in 71 patients (94%) with a median value of 464 mAU/Ml (range 27–40783 mAU/mL); the sensitivity and specificity were 78.67% and 90.67% respectively. Patients with pre-operative PIVKA-II positivity showed a significant decrease (P < 0.015) of median PIVKA-II serum concentrations after surgery: 820 (91–40783) mAU/mL at diagnosis vs 123 (31–4666) mAU/mL post-operatively. IF assay on PDAC sections demonstrated PIVKA-II expression in cancer cells.ConclusionThese data are the first showing a decreased PIVKA-II serum levels after surgery in PDAC patients and reporting PIVKA-II expression in PDAC tissue. Further studies are needed to confirm these findings and to determine PIVKA-II usefulness in diagnosing and monitoring PDAC patients.

Development of a New Chemiluminescent Enzyme Immunoassay Using a Two-Step Sandwich Method for Measuring Aldosterone Concentrations.

In the present study, we developed a new chemiluminescent enzyme immunoassay (CLEIA) using a two-step sandwich method to measure aldosterone concentrations. We investigated serum and plasma aldosterone concentrations in 75 blood samples from 27 patients using a radioimmunoassay (RIA) and the CLEIA (with current and newly improved reagents) as well as liquid chromatography-tandem mass spectrometry (LC-MS/MS). Based on the results of the Passing–Bablok regression analysis, the aldosterone levels measured using CLEIA with the new reagents and those measured by LC-MS/MS were found to be significantly correlated (slope, 0.984; intercept, 0.2). However, aldosterone levels varied depending on the measurement method (i.e., CLEIA with the new reagent, CLEIA with the current reagent, and RIA). Aldosterone levels were lower with the improved CLEIA method than with RIA and CLEIA using the current reagent. Therefore, the cutoff values of the screening test as well as those of the confirmatory test for primary aldosteronism (PA) should be adjusted to follow current clinical practice guidelines for PA. The formula that can be used to obtain the aldosterone level (pg/mL) when using CLEIA with the new reagent is 0.765 × RIA (pg/mL) − 33.7. This formula will enable PA cutoff values to be set for provisional screening and confirmatory tests.

Prevalence of total hepatitis A\xa0antibody among 5 to 7\xa0years old children and their mothers in Cambodia

This study determined the prevalence of total hepatitis A antibody (anti-HAV) among 5–7 years old children and their mothers in the whole Cambodia, using a nationwide study, and examined the differences between the two cohorts. A total of 4535 dried blood spot-driven (DBS) samples (2021 mothers and their 2514 children of 5–7 years old) and the concomitant 922 whole blood samples (subset of the whole participants) were collected using a multistage random sampling strategy throughout Cambodia in 2017. Total anti-HAV was detected using the chemiluminescence enzyme immunoassay method. Compared to gold standard whole blood samples, the sensitivity and specificity of DBS mediated anti-HAV detection were 94.8% and 98%, respectively. Total anti-HAV prevalence among mothers was 91.2% (95%CI: 90.0–92.5%), and that of their children was 31.5% (95%CI: 29.7–33.3%). In our study, the low prevalence of total anti-HAV among children indicates the country’s improvement of safe water and food supply, hygiene and sanitation. If the hygiene and sanitation are consistently improved in Cambodia, the prevalence might be no longer increased when the children become adults.

Evaluation of Toxoplasma gondii IgM and IgG Seropositivities in Serum Samples Sent from Pediatric and Adult Hematology/Oncology Outpatient Clinics

Toxoplasmosis, despite its mild course in immunocompetent individuals, it may have a more severe course in immunosuppressed patients such as hematology and oncology patients. This study aimed to contribute to the importance of that issue by evaluating the seropositivity rates of Toxoplasma gondii (T. gondii) in Hematology and Oncology patients and sharing the data obtained.&#x0D; Serum samples sent from pediatric and adult Hematology/Oncology outpatient clinic to the Microbiology Laboratory between January 2017 and August 2019 were analysed using Architect Toxo IgG and IgM Reagent Kit with chemiluminescent microparticular enzyme immunoassay method on Architect system.&#x0D; A total of 673 patient samples belonging to 131 pediatric patients and 542 adult patients were analysed in the study. Median age of the study group was 29 (range: 0 - 83) and 53.5% of them were male. While T. gondii IgM positivity was 8% IgG positivity was 28.7%. T. gondii IgG seropositivities in male and female patients were 27.2% and 30.4% respectively (p = 0.44). T. gondii IgM seropositivities in pediatric and adult patients were 6.9% and 8.3% respectively and IgG seropositivities were 24.4% and 29.7% (p = 0.78 and p = 0.46 respectively). T. gondii IgM positivity was found as 8% and IgG positivity as 28.7% in the study group. In conclusion, data obtained from the study were in compliance with both domestic and foreign data and the highest rate of IgM and IgG seropositivity was found in patients with lymphoma.

To measure and compare the serum levels of homocysteine in Pre-eclamptic and normotensive pregnancies in a tertiary care hospital

Materials and Methods: The present prospective case control study was conducted in the Department of Obstetrics and Gynecology, SVS Medical College and Hospital, MBNR. Pregnant women with preeclampsia were considered as cases and women without any medical or other obstetric and fetal complications were selected as controls. Serum homocysteine was estimated on 2 ml of serum by competitive chemiluminescent enzyme immunoassay method.

Diagnostic performance of Mac-2 binding protein glycosylation isomer (M2BPGi) in screening liver fibrosis in health checkups.

BackgroundMild‐to‐moderate fibrosis is rarely diagnosed because the disease is asymptomatic in the early stage. The serum level of Mac‐2 binding protein glycosylation isomer (M2BPGi) has been found to increase with the severity of liver fibrosis. The aim of this study was to determine the diagnostic performance of M2BPGi in screening liver fibrosis using magnetic resonance elastography (MRE) as a reference standard and to compare it with using the aspartate aminotransferase‐to‐platelet ratio (APRI) and the Fibrosis‐4 index (FIB‐4) in health checkups.MethodsThis cross‐sectional study consecutively selected subjects at health examinations who underwent MRE and M2BPGi testing at eight health promotion centers in Korea between January and September 2019. The serum M2BPGi level was measured using the chemiluminescence enzyme immunoassay method. The measured levels were indexed using the cutoff index (COI). COI values of M2BPGi were compared with the MRE results.ResultsThe median (interquartile) values of COI for fibrosis stages F0 (normal liver stiffness), F1 (mild fibrosis), F2 (significant fibrosis), and ≥F3 (advanced fibrosis) were 0.49 (0.34‐0.61), 0.48 (0.38‐0.68), 0.64 (0.43‐1.03), and 1.01 (0.75‐1.77), respectively (P < .0001). The AUCs of the COI for the screening of fibrosis stage ≥F1, ≥F2, and ≥F3 were 0.591, 0.698, and 0.853, respectively. Using a threshold of 0.75 for COI to exclude advanced fibrosis had a sensitivity, specificity, and negative predictive value of 80.0%, 77.9%, and 98.9%, respectively. The AUC for excluding advanced fibrosis was better for M2BPGi than for FIB‐4 and APRI.ConclusionSerum M2BPGi was useful for screening significant and advanced fibrosis in health checkups.

Clinical significance of serum PSA in breast cancer patients

BackgroundRecent preclinical data suggest that androgen receptor (AR) signaling plays a significant role in subsets of breast cancer. Clinical trials testing AR-targeting therapies in breast cancer have been conducted. Assessment of AR-signal in breast cancer tissue maybe useful for treatment selections. Prostate specific antigen (PSA) is the product of an androgen-responsive gene. Serum PSA (sPSA) can be detected in women by a highly sensitive assay although the concentration is much lower than that observed in males. We investigated if sPSA reflects tumor biology, including AR signaling in breast cancer patients.MethodsIn this study, 132 healthy controls and 144 breast cancer patients were enrolled. sPSA was evaluated by the chemiluminescent enzyme immunoassay (CLEIA) method. Correlations between sPSA and the various clinicopathological factors were analyzed.ResultsIn post-menopausal state, sPSA detection rate was significantly higher in breast cancer patients compared with controls (27.4% vs 11.3%: p = 0.0090), but not in the whole cohort (29.2% vs 25.8%: p = 0.5265) or pre-menopausal subgroup (37.0% vs 42.6%: p = 0.6231). In post-menopausal breast cancer cases, higher sPSA value was associated with clinic-pathological factors including the expression of AR protein in primary legion. In a correlation analysis of quantitative data limited to post-menopausal metastatic breast cancer (MBC), sPSA was positively, albeit weakly correlated with clinic-pathological features including serum testosterone levels and AR positivity.ConclusionsOur data suggest that sPSA may reflect tumor biological properties including AR activity in post-menopausal breast cancer.