Chemiluminescence Immunoassay Kit Research Articles

265 PROGNOSTIC ROLE OF VITAMIN D DEFICIENCY IN STEMI PATIENTS UNDERGOING PRIMARY PERCUTANEOUS CORONARY INTERVENTION

Abstract Background and Aims The pandemic proportion of vitamin D deficiency has raised particular concerns on its association with mortality and cardiovascular disease (CVD). However, the independent and causative role of hypovitaminosis D in CVD is still debated. Few data have been reported, so far, among STEMI patients undergoing primary percutaneous coronary intervention (pPCI), that was therefore the aim of the present study. Methods A consecutive cohort of patients admitted for STEMI treated with pPCI were included. The levels of 25(OH)D were assessed at admission by chemiluminescence immunoassay kit LIAISON® Vitamin D assay (Diasorin Inc). Hypovitaminosis D was defined for 25(OH)D < 10 ng/ml. The primary study endpoint was net adverse cardiovascular events (a composite of death, recurrent MI and target vessel revascularization) at the longest available follow-up. Secondary endpoints were overall mortality, major bleedings or the individual components of the primary endpoint. Results We included 136 patients divided according to tertiles values of 25(OH)D (< 12.98 ng/ml, n=44; 12.98-22.1 ng/ml, n=41; ≥ 22.1 ng/ml, n=51). The prevalence of renal failure (p=0.047) and coronary calcifications inversely related with vitamin D. Lower vitamin D levels were associated with “no reflow” (10.3% vs 6.7% vs 0%, p=0.04), with increased use of adenosine. At a mean follow-up of 305± 217 days (mean + SD), the primary endpoint (NACE) was slightly increased in patients with lower vitamin D values (adjusted HR [95% CI] = 3.07 [0.85-11.1], p = 0.09). Similar results were observed for ischemic events (MACE: adjusted HR [95% CI] = 2.78 [0.68-11.48], p = 0.16). However, when considering separately patients with severe hypovitaminosis D (<10 ng/ml), we found a significant increase in the risk of NACE and MACE (adjusted HR [95% CI] = 5.17 [1.42-18.8], p = 0.01 and HR = 4.75 [1.14-19.7], p = 0.03, respectively). Conclusion The present study shows that among patients with STEMI undergoing pPCI, extremely low levels of vitamin D are independently associated with impaired outcomes. Future dedicated studies will shed light on the prognostic implications of hypovitaminosis D in these patients and the potential therapeutic perspectives.

Pediatric Continuous Reference Intervals of Serum Insulin-like Growth Factor 1 Levels in a Healthy Chinese Children Population – Based on PRINCE Study

ObjectivesWe aimed to establish age- and sex-dependent reference intervals for insulin-like growth factor 1 (IGF-1) based on the measurements of healthy Chinese children from the pediatric reference intervals in China study and to investigate whether body mass index (BMI) and height affect IGF-1 levels. MethodsA total of 3753 individuals with eligible blood specimens resampled from the pediatric reference intervals in China population were enrolled as reference individuals. IGF-1 levels were measured using a chemiluminescent immunoassay kit. The lower limit and upper limit values of the reference individuals were calculated by defining the 2.5th and 97.5th percentiles. The skewness-median-coefficient of variation method was used to calculate the standard deviation score (SDS) of serum IGF-1, and cubic spline curves were applied to depict a smoothed curve for each age- and sex-specific stratification of the L, M, and S parameters. ResultsSerum IGF-1 levels increased with age from the age of 1 year, peaking at around the age of 13 years in girls and 15 years in boys and then began to decline (both P <.001). Before 14 years, IGF-1 levels were higher in girls than in boys at the same age, and the difference was statistically significant (P < .05), but there was no significant difference in the IGF-1 levels between girls and boys aged 14 to 16 and 18 years. The Spearman correlation coefficients of height SDS, weight SDS, and BMI SDS with IGF-1 SDS were 0.29, 0.33, and 0.20, respectively (P < .001). ConclusionThis study established age- and sex-specific normative IGF-1 data for Chinese children and adolescents between the ages of 1 and 19 years. The BMI and height SDS had no effect on IGF-1 levels in healthy children.

BP180NC16a autoantibody positivity may predict low-risk mucous membrane pemphigoid.

The objective of the present study was to explore the clinical characteristics of BP180NC16a autoantibody-positive mucous membrane pemphigoid. Data from 22 patients were collected by retrospective chart review. For the detection of BP180NC16a autoantibody, a commercially available enzyme-linked immunosorbent assay or chemiluminescent enzyme immunoassay kit was used. The patients were classified into the following clinical phenotypes: low-risk patients who had lesions only in the oral mucosa or in the oral mucosa and skin and high-risk patients who had lesions in the oral mucosa and any of the following sites: the eyes, upper respiratory tract, or esophagus. Eleven of the 22 patients (50%) were BP180NC16a-positive at the time of diagnosis. All the positive patients were in the low-risk group. There was a significant difference in the incidence of low-risk mucous membrane pemphigoid between BP180NC16a autoantibody-positive and BP180NC16a autoantibody-negative patients (p=0.004). Patients' age, disease duration, and oral disease activity scores were not significantly different between the two groups. BP180NC16a reactivity was associated with the clinical phenotype characterized by only oral mucosal lesions or combined oral mucosal and skin lesions. BP180NC16a autoantibody might be useful as a serum marker to predict low-risk mucous membrane pemphigoid.

Detection of Circulating VZV-Glycoprotein E-Specific Antibodies by Chemiluminescent Immunoassay (CLIA) for Varicella-Zoster Diagnosis.

Varicella and herpes zoster are mild symptoms-associated diseases caused by varicella–zoster virus (VZV). They often cause severe complications (disseminated zoster), leading to death when diagnoses and treatment are delayed. However, most commercial VZV diagnostic tests have low sensitivity, and the most sensitive tests are unevenly available worldwide. Here, we developed and validated a highly sensitive VZV diagnostic kit based on the chemiluminescent immunoassay (CLIA) approach. VZV-glycoprotein E (gE) was used to develop a CLIA diagnostic approach for detecting VZV-specific IgA, IgG, and IgM. The kit was tested with 62 blood samples from 29 VZV-patients classified by standard ELISA into true-positive and equivocal groups and 453 blood samples from VZV-negative individuals. The diagnostic accuracy of the CLIA kit was evaluated by receiver-operating characteristic (ROC) analysis. The relationships of immunoglobulin-isotype levels between the two groups and with patient age ranges were analyzed. Overall, the developed CLIA-based diagnostic kit demonstrated the detection of VZV-specific immunoglobulin titers depending on sample dilution. From the ELISA-based true-positive patient samples, the diagnostic approach showed sensitivities of 95.2%, 95.2%, and 97.6% and specificities of 98.0%, 100%, and 98.9% for the detection of VZV-gE-specific IgA, IgG, and IgM, respectively. Combining IgM to IgG and IgA detection improved diagnostic accuracy. Comparative analyses on diagnosing patients with equivocal results displaying very low immunoglobulin titers revealed that the CLIA-based diagnostic approach is overall more sensitive than ELISA. In the presence of typical VZV symptoms, CLIA-based detection of high titer of IgM and low titer of IgA/IgG suggested the equivocal patients experienced primary VZV infection. Furthermore, while no difference in IgA/IgG level was found regarding patient age, IgM level was significantly higher in young adults. The CLIA approach-based detection kit for diagnosing VZV-gE-specific IgA, IgG, and IgM is simple, suitable for high-throughput routine analysis situations, and provides enhanced specificity compared to ELISA.

Relationship between vitamin D and cholesterol levels in STEMI patients undergoing primary percutaneous coronary intervention

Background and aimsSpecial interest has been raised on vitamin D association with the metabolic profile, potentially interfering with lipid parameters and lipid-lowering therapies. The aim of the present study was to assess the impact of vitamin D on the cholesterol levels among patients with ST-segment elevation myocardial infarction (STEMI) undergoing primary PCI. Methods and resultsA consecutive cohort of 450 patients admitted for STEMI treated with pPCI were retrospectively identified and divided according to tertiles values of 25(OH). The levels of 25(OH)D were assessed at admission by chemiluminescence immunoassay kit LIAISON®Vitamin D assay (Diasorin Inc).Lower vitamin D was associated to a higher use of diuretics (p = 0.03), lower prevalence of lesions on bifurcations (p = 0.001) and smaller diameter of the target coronary vessel (p = 0.03), but higher coronary calcifications (p = 0.007). Total and LDL cholesterol levels were significantly increased in patients with lower vitamin D (p = 0.05 and p = 0.005), inversely relating with total cholesterol (r = −0.09, p = 0.06) and LDL-C (r = −013, p = 0.007), and directly with HDL-C (r = 0.16, p = 0.001). Results were not affected by statin therapy, with a significant relationship being confirmed for atherogenic lipids, but not for HDL-C in statin treated patients.In fact, at multivariate analysis, vitamin D in lower tertiles emerged as an independent predictor of LDL-C elevated or above the target (adjusted OR [95%CI] = 2.6 [1.51–4.44], p = 0.001). ConclusionThe present study shows that among patients with STEMI undergoing primary revascularization, lower levels of vitamin D are independently associated with a more atherogenic lipid profile. Similar results were observed in statin treated or naïve patients.

Randomized Controlled Study of the Effects of DHEA on the Outcome of IVF in Endometriosis.

Objective To investigate the effect of dehydroepiandrosterone (DHEA) on the outcome of in vitro fertilization (IVF) in patients with endometriosis (EMT). Methods Female patients diagnosed with EMT in our hospital from May 2018 to May 2019 were selected. The patients were divided into the control group (n = 22) and the DHEA group (n = 22) according to the random number table. Patients in the control group received placebo and patients in the DHEA group received DHEA. Patients in both groups received either DHEA (25 mg) or placebo orally 3 times a day for 90 days from the first day of menstruation. Patients were subsequently treated with an IVF cycle. In the control group, 22 patients completed the first cycle and 13 patients completed the second cycle. In the DHEA group, 22 patients completed the first cycle and 11 patients completed the second cycle. Serum sex hormone levels including serum E2 on hCG day, mean progesterone on hCG day, FSH on day 2, AMH on day 2, and gonadotropin dose were determined using a chemiluminescent immunoassay kit. The number of antral follicles of the bilateral ovaries was counted by transvaginal B-ultrasound, and the maximum length and transverse diameter of the ovaries were measured at the same time, to calculate the average diameter of the ovaries, observe the morphology of endometrium, and measure the thickness of the endometrium. The implantation rate, clinical pregnancy rate, persistent pregnancy rate, and live birth rate were compared between the two groups. Results There were no significant differences in serum E2, progesterone, endometrial thickness, recovered oocytes, mean number of transferred embryos, and mean score of leading embryo transfer between the DHEA group and the women who completed the first and second cycles (P > 0.05). The AMH, antral follicle count, serum E2 on hCG day, the number of recovered oocytes, fertilized oocytes, and the fertilization rate in the DHEA group were higher than those in the control group (P < 0.05). The doses of FSH on day 2, COH on day 3, and gonadotropin were lower than those in the control group (P < 0.05). There was no significant difference in the total number of embryos, the number of high-quality embryos, and the number of transplanted embryos between the two groups (P > 0.05). The implantation rate, clinical pregnancy rate, persistent pregnancy rate, and live birth rate in the DHEA group were higher than those in the control group (P < 0.05). Conclusion DHEA can significantly increase serum E2 level and improve IVF outcome by regulating the hormone synthesis process, thus improving oocyte and embryo quality.

Vitamin D deficiency predicts the no-reflow phenomenon in STEMI patients undergoing primary percutaneous coronary intervention

Abstract Background Vitamin D displays a broad spectrum of cardioprotective effects, preventing oxidative stress, inflammation and thrombosis and improving endothelial function. Previous studies have associated vitamin D deficiency with more extended and severe coronary artery disease (CAD) and worse outcome, and especially among patients with ST-segment elevation myocardial infarction (STEMI). Few data have been reported on the association of vitamin D levels with the features of infarct-related lesions and PCI outcomes. Aim We aimed at assessing the relationship between vitamin D and angiographic findings and the procedural results of primary percutaneous coronary intervention (pPCI) in STEMI. Methods A consecutive cohort of patients admitted for STEMI treated with pPCI were included. The levels of 25(OH)D were assessed at admission by chemiluminescence immunoassay kit LIAISON® Vitamin D assay. Hypovitaminosis D was defined for 25(OH)D &amp;lt;10 ng/ml. Results We included in our study 450 patients, divided according to tertiles values of 25(OH)D. Lower vitamin D was associated to higher use of diuretics (p=0.02), higher levels of white blood cells and glycemia (p&amp;lt;0.001), lower prevalence of lesions on bifurcations (p=0.03) and smaller diameter of the target coronary vessel (p=0.03). Procedural characteristics and pre-procedural TIMI flow were not different according to vitamin D levels, but for a higher rate of slow-flow/no-reflow phenomenon (12.8% vs 8.1% vs 5.3%, p=0.03, adjusted OR [95% CI]=2.6 [1.05–6.6], p=0.04 for I vs III tertile), requiring higher use of adenosine (p=0.006) and glycoprotein IIbIIIa inhibitors (p=0.01). Conclusion The present study shows that among patients with STEMI undergoing pPCI, lower levels of vitamin D indepently predict the occurrence of slowflow/no-reflow phenomenon. Future dedicated studies will shed light on the prognostic implications of hypovitaminosis D in these patients and the potential therapeutic perspectives. Funding Acknowledgement Type of funding sources: None.

Performance evaluation of automated chemiluminescence immunoassay based antigen detection – Moving towards more reliable ways to predict SARS-CoV-2 infection

Real-time reverse transcription- polymerase chain reaction (RT-PCR) has been the most reliable armoury for the diagnosis of COVID-19, considered to be the reference standard but fails to reproduce the correct predictability about the infectivity of the disease every time. Antigen detection however puts foothold in this aspect even though lacks in sensitivity, especially conventional Rapid Antigen Tests (RATs). Recently developed Chemiluminescence Immunoassay (CLIA) based antigen detection tests are promising and displayed better sensitivity. In the current study we have evaluated VITROS® SARS-CoV-2 Ag Test CLIA Kit, which was tested on 148 patient’s samples attended to a tertiary care centre for testing of SARS-CoV-2. The performance of the kit was evaluated in comparison to RT-PCR and RAT and found to be a good test for antigen detection, best within the first few days of infection. The test has shown sensitivity of 94.3 % and specificity of 100 % in samples with corresponding Ct values of ≤25 by RT-PCR, which corresponds to high viral load and can predict ability of spreading the disease by the patients. With the results being semiquantitative along with improved sensitivity it can replace RATs for antigen detection for screening, provided good laboratory set up is included under consideration.

SARS-CoV-2 Antibodies and Associated Factors at Different Hospitalization Time Points in 192 COVID-19 Cases.

BackgroundWe launched a retrospective analysis of SARS-CoV-2 antibodies in 192 patients with COVID-19, aiming to depict the kinetic profile of SARS-CoV-2 antibodies and explore the factors related to SARS-CoV-2 antibody expression.MethodsData on 192 confirmed patients with COVID-19 between January and February 2020 was collected from the designated hospital that received patients with COVID-19 in Guangzhou, China. Moreover, a cohort of 130 suspected patients with COVID-19 and 209 healthy people were also enrolled in this study. IgM and IgG antibodies to SARS-CoV-2 were detected by the chemiluminescence immunoassay kits in different groups.ResultsA total of 192 COVID-19 cases were analyzed, of which had 81.8% anti-SARS-CoV-2 IgM detected and 93.2% anti-SARS-CoV-2 IgG detected, respectively, at the time of sampling. The kinetics of anti-SARS-CoV-2 IgM and IgG showed that, the confirmed cases had anti-SARS-CoV-2 IgM seroconversion occurred 5–10 days after the onset of the symptoms, and then IgM rose rapidly to reach a peak within around 2–3 weeks, maintaining at its peak for 1 week before its decline. While they had anti-SARS-CoV-2 IgG seroconversion simultaneously or sequentially with IgM, reaching its peak within around 3 to 4 weeks and began to decline after the fifth week. Besides, correlation analysis showed that in patients with COVID-19 the level of IgM was related to gender and disease severity (P < 0.01), and the level of IgG was related to age and disease severity (P < 0.001). The univariate analysis of relevant factors indicated that the level of IgG had a weak correlation with age (r = 0.374, P < 0.01). The level of IgM in male patients was higher than that in female patients (P < 0.001). The expression level of anti-SARS-CoV-2 IgM and IgG were positively correlated with the severity of COVID-19 and the duration of the virus in the patients.ConclusionThe findings of this study show that anti-SARS-CoV-2 IgM and IgG can be important assisting COVID-19 diagnosis, especially in the early phase of infection. Furthermore, antibody expression in patients with COVID-19 is also correlated with disease severity, age, gender, and virus clearance or continuous replication.

Vitamin D deficiency is associated with impaired reperfusion in STEMI patients undergoing primary percutaneous coronary intervention

Background and aimsVitamin D displays a broad spectrum of cardioprotective effects, preventing oxidative stress, inflammation and thrombosis and improving endothelial function.Previous studies have associated vitamin D deficiency with more extended and severe coronary artery disease (CAD) and worse outcome, and especially among patients with ST-segment elevation myocardial infarction (STEMI). However, few data have been reported on the association of vitamin D levels with the angiographic findings and epicardial reperfusion in STEMI patients undergoing primary percutaneous coronary intervention (pPCI), that was therefore the aim of the present study. Methods and resultsA consecutive cohort of patients admitted for STEMI and treated with pPCI were included. The levels of 25(OH)D were assessed at admission by chemiluminescence immunoassay kit LIAISON® Vitamin D assay (Diasorin Inc). Hypovitaminosis D was defined for 25(OH)D < 10 ng/ml.We included in our study 450 patients, divided according to tertiles values of 25(OH)D. Lower vitamin D was associated to a higher use of diuretics (p = 0.02), higher levels of white blood cells and glycemia (p < 0.001), lower prevalence of lesions on bifurcations (p = 0.03) and smaller diameter of the target coronary vessel (p = 0.03). Procedural characteristics and pre-procedural TIMI flow were not different according to vitamin D levels, but for a higher rate of impaired epicardial reperfusion (12.8% vs 8.1% vs 5.3%, p = 0.03, adjusted OR[95%CI] = 2.6[1.05–6.6], p = 0.04 for I vs III tertile), requiring higher use of adenosine (p = 0.006) and glycoprotein IIbIIIa inhibitors (p = 0.01). ConclusionThe present study shows that among patients with STEMI undergoing pPCI, lower levels of vitamin D are independently associated with impaired reperfusion, Future dedicated studies will shed light on the prognostic implications of hypovitaminosis D in these patients and the potential therapeutic perspectives.

Comparison of Serum Thyroid Hormone Levels in Green-winged Macaws (Ara chloropterus) Using Radio and Chemiluminescent Immunoassays.

This study compared the use of radioimmunoassay (RIA) and chemiluminescent immunoassay (CLIA) to measure serum thyroid hormone levels in green-winged macaws (Ara chloropterus). The sample group comprised 20 male and 13 female (n = 33) healthy, captive green-winged macaws of reproductive age but outside of breeding season. Mean free triiodothyronine (T3), total T3, and free thyroxine (T4) measured by CLIA and RIA corresponded to 5.11 and 5.08 pmol/L (3.33 and 3.31 pg/mL), 1.59 and 1.56 nmol/L (103.5 and 101.5 ng/dL), and 8.25 and 8.82 pmol/L (0.64 and 0.69 ng/dL), respectively. Mean total T4 by RIA corresponded to 6.29 nmol/L (0.49 µg/dL). Mean free T4 and thyroid-stimulating hormone (TSH) levels differed according to immunoassay method, with higher values measured by RIA compared with CLIA. Sex influence was statistically significant in immunoassay results only on free T3 levels. Free T3 levels measured by RIA were higher than levels measured by CLIA in male birds, but this difference was not found with the female bird samples. Conversely, free T3 levels measured by CLIA were higher in the female macaws compared with male birds. Comparative analysis of thyroid hormone measurements in this study revealed that RIA and CLIA are equivalent methods to measure free T4 and total T3 levels but not TSH levels. These findings support the use of CLIA for free T4 and total T3 level determinations in green-winged macaws. However, the CLIA kit used in this study provided invalid total T4 level results for the macaws sampled. Radioimmunoassay and CLIA were equally ineffective for determining TSH levels in this species.

Performance Verification of Plasma Renin and Aldosterone Examination with Chemiluminescence Immunoassay and Its Screening Efficacy for Primary Aldosteronism

To evaluate the performance of chemiluminescence immunoassay (CLIA) in examining renin and aldosterone and to determine its value for screening for primary aldosteronism (PA). According to the relevant documents of Clinical and Laboratory Standards Institute (CLSI), we verified the precision, linear range and carryover rate of examining renin and aldosterone with CLIA. The study included 91 suspected PA patients, using two methods, CLIA and radioimmunoassay (RIA), to examine renin and aldosterone levels in order to compare the correlation between the two methods and their value for PA screening. The precision, linear range and carryover rate of examining renin and aldosterone with CLIA met the requirements. In patients with suspected PA, the correlation coefficients of renin, aldosterone and aldosterone-to-renin ratio (ARR) assessed by CLIA and RIA were 0.901, 0.861 and 0.847 respectively (all P<0.001). When the patients were in the upright position and the ARR was 5.636 (ng/dL)/(ng/L), the CLIA method had 79.1% sensitivity and 93.7% specificity for PA screening; when ARR was 14.084 (ng·dL －1)/(ng·[mL·h] －1), the RIA method had 93.0% sensitivity and 83.3% specificity for PA screening. When the patients were in the supine position, and the ARR was 5.640 (ng/dL)/(ng/L), the CLIA method had 97.7% sensitivity and 81.2% specificity for PA screening; when ARR was 33.494 (ng·dL －1)/(ng·[mL·h] －1), RIA had 95.3% sensitivity and 70.8% specificity for PA screening . The performance of the CLIA kit in assessing the concentration of renin and aldosterone meets the clinical requirements. Regarding preliminary PA screening, upright-position ARR had higher specificity, but lower sensitivity compared with supine-position ARR.

Development of a Nucleocapsid Protein-Based ELISA for Detection of Human IgM and IgG Antibodies to SARS-CoV-2.

SARS-CoV-2 is the etiologic agent of COVID-19, which has led to a dramatic loss of human life and presents an unprecedented challenge to public health worldwide. The gold standard assay for SARS-CoV-2 identification is real-time polymerase chain reaction; however, this assay depends on highly trained personnel and sophisticated equipment and may suffer from false results. Thus, a serological antibody test is a supplement to the diagnosis or screening of SARS-CoV-2. Here, we develop and evaluate the diagnostic performance of an IgM/IgG indirect ELISA method for antibodies against SARS-CoV-2 in COVID-19. The ELISA was constructed by coating with a recombinant nucleocapsid protein of SARS-CoV-2 on an enzyme immunoassay plate, and its sensitivity and specificity for clinical diagnosis of SARS-CoV-2 infection was assessed by detecting the SARS-CoV-2-specific IgM and IgG antibodies in COVID-19 patient’s sera or healthy person’s sera. The SARS-CoV-2 positive serum samples (n = 168) were collected from confirmed COVID-19 patients. A commercial nucleocapsid protein-based chemiluminescent immunoassay (CLIA) kit and a colloidal gold immunochromatography kit were compared with those of the ELISA assay. The specificity, sensitivity, positive predictive value (PPV), and negative predictive value (NPV) of IgM were 100, 95.24, 100, and 91.84%, whereas those of IgG were 100, 97.02, 100, and 94.74%, respectively. We developed a highly sensitive and specific SARS-CoV-2 nucleocapsid protein-based ELISA method for the diagnosis and epidemiologic investigation of COVID-19 by SARS-CoV-2 IgM and IgG antibody detection.

Development of quantum dot-based fluorescence lateral flow immunoassay strip for rapid and quantitative detection of serum interleukin-6.

BackgroundInterleukin‐6 (IL‐6) is an inflammatory factor that increases rapidly in response to infectious diseases including sepsis. The aim of this study is to develop a quantum dot (QD)‐based fluorescence lateral flow immunoassay (LFIA) strip that can rapidly and accurately detect IL‐6 levels.MethodsQD‐based LFIA strips were fabricated by conjugating CdSe/ZnS QDs to the IL‐6 antibody. Performance verification and clinical sample analysis were carried out to evaluate the newly developed strip.ResultsQD‐based LFIA strips were successfully fabricated. The test strip's linear range was 10–4000 pg/ml, with a linear correlation coefficient of R 2 ≥ .959. The sensitivity of the test strip was 1.995 pg/ml. The recovery rate was 95.72%–102.63%, indicating satisfying accuracy. The coefficient of variation (CV) of the intra‐assay was 2.148%–3.903%, while the inter‐assay was 2.412%–5.293%, verifying the strip's high precision. The cross‐reaction rates with various interleukins (IL‐1α, IL‐1β, IL‐2, IL‐4, and IL‐8) and interferon‐γ (IFN‐γ) were all <0.1%. When the strip was placed in a 50°C oven for 1, 2, 3, and 4 weeks, the test results were not significantly altered compared to storage at room temperature. Furthermore, 200 clinical serum samples were analyzed to compare the strip with the Beckman chemiluminescence immunoassay (CLIA) kit, which revealed a high correlation (n = 200, R 2 = .9971) for the detection of IL‐6.ConclusionsThe QD‐based test strip can rapidly and quantitatively detect IL‐6 levels, thus meeting the requirement of point‐of‐care test (POCT) and showing excellent clinical prospects.

Point-of-care test system for detection of immunoglobulin-G and -M against nucleocapsid protein and spike glycoprotein of SARS-CoV-2.

The coronavirus disease 2019 (COVID-19) epidemic continues to ravage the world. In epidemic control, dealing with a large number of samples is a huge challenge. In this study, a point-of-care test (POCT) system was successfully developed and applied for rapid and accurate detection of immunoglobulin-G and -M against nucleocapsid protein (anti-N IgG/IgM) and receptor-binding domain in spike glycoprotein (anti-S-RBD IgG/IgM) of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). Any one of the IgG/IgM found in a sample was identified as positive. The POCT system contains colloidal gold-based lateral flow immunoassay test strips, homemade portable reader, and certified reference materials, which detected anti-N and anti-S-RBD IgG/IgM objectively in serum within 15 min. Receiver operating characteristic curve analysis was used to determine the optimal cutoff values, sensitivity, and specificity. It exhibited equal to or better performances than four approved commercial kits. Results of the system and chemiluminescence immunoassay kit detecting 108 suspicious samples had high consistency with kappa coefficient at 0.804 (P < 0.001). Besides, the levels and alterations of the IgG/IgM in an inpatient were primarily investigated by the POCT system. Those results suggested the POCT system possess the potential to contribute to rapid and accurate serological diagnosis and epidemiological survey of COVID-19.

Association of lower vitamin D levels with inflammation and leucocytes parameters in patients with and without diabetes mellitus undergoing coronary angiography.

Diabetes mellitus has been associated with a chronic low-grade inflammation and a higher risk of cardiovascular and infectious disease, that could be prevented by the effects of vitamin D. We aimed at evaluating the impact of vitamin D levels on the biomarkers of acute-phase response, inflammation and glucose metabolism in a large cohort of diabetic patients with cardiovascular disease. Consecutive patients undergoing coronary angiography were included. Diabetes mellitus was defined as previous diagnosis, specific treatment administration (oral drug or insulin), fasting glycaemia >6.99mmol/L or HbA1c >48mmol/L. Glucose parameters, white blood cells, Neutrophil-to-Lymphocyte Ratio (NLR), Monocyte-to-Lymphocyte Ratio (MLR), C-reactive protein (CRP) and vitamin D were measured at admission. Vitamin D levels were measured by chemiluminescence immunoassay kit LIAISON® Vitamin D assay (Diasorin Inc). We included 1472 diabetic patients and 2499 non-diabetic patients that were divided according to vitamin D tertiles. Among diabetic patients, lower levels of vitamin D were associated with female gender (P=.02), obesity (P=.004), active smoking and acute presentation (P<.001) and with a more atherogenic metabolic profile. The levels of white blood cells, leucocytes subfamilies, and inflammatory parameters significantly correlated with vitamin D levels in both patients with and without diabetes (diabetic: P=.012 for WBC, P=.004 for NLR and P<.001 for MLR and C-reactive protein, non-diabetic: P<.001 for WBC; NLR, MLR and C-reactive protein, respectively). Among diabetic patients, results were confirmed at multivariate analysis with no significant interaction according to glycaemic control. The present study demonstrates that, among patients with cardiovascular disease, vitamin D deficiency is associated with metabolic dysregulation and with an elevation of cellular and humoural inflammatory parameters, especially among diabetics, although not being dependent from glycaemic control.

Predictive Value of Serum Ferritin in Combination with Alanine Aminotransferase and Glucose Levels for Noninvasive Assessment of NAFLD: Fatty Liver in Obesity (FLiO) Study.

The identification of affordable noninvasive biomarkers for the diagnosis and characterization of nonalcoholic fatty liver disease (NAFLD) is a major challenge for the research community. This study aimed to explore the usefulness of ferritin as a proxy biomarker of NAFLD condition, alone or in combination with other routine biochemical parameters. Subjects with overweight/obesity and ultrasound-confirmed liver steatosis (n = 112) from the Fatty Liver in Obesity (FLiO) study were assessed. The hepatic evaluation considered magnetic resonance imaging, ultrasonography, and credited routine blood liver biomarkers. Anthropometry and body composition, dietary intake (by means of a validated 137-item food frequency questionnaire), and specific biochemical markers were also determined. Serum ferritin levels were analyzed using a chemiluminescent microparticle immunoassay kit. Lower serum ferritin concentrations were associated with general better liver health and nutritional status. The evaluation of ferritin as a surrogate of liver damage by means of quantile regression analyses showed a positive association with alanine aminotransferase (ALT) (β = 19.21; p ≤ 0.001), liver fat content (β = 8.70; p = 0.008), and hepatic iron (β = 3.76; p ≤ 0.001), after adjusting for potential confounders. In receiver operating characteristic (ROC) analyses, the panel combination of blood ferritin, glucose, and ALT showed the best prediction for liver fat mass (area under the curve (AUC) 0.82). A combination of ferritin and ALT showed the higher predictive ability for estimating liver iron content (AUC 0.73). This investigation demonstrated the association of serum ferritin with liver health as well as with glucose and lipid metabolism markers in subjects with NAFLD. Current findings led to the identification of ferritin as a potential noninvasive predictive biomarker of NAFLD, whose surrogate value increased when combined with other routine biochemical measurements (glucose/ALT).

Cholecalciferol levels, inflammation and leukocytes parameters: Results from a large single-centre cohort of patients

Vitamin D deficiency represents a pandemic health problem with a broad spectrum of clinical implications. Several studies have involved lower levels of vitamin D with inflammatory disorders including cardiovascular, autoimmune and infectious disease. Indeed, the pathophysiological mechanisms are still poorly ascertained. We aimed at evaluating the impact of cholecalciferol (25(OH)D) levels on the biomarkers of acute-phase response and inflammation in a large cohort of patients with cardiovascular disease. Consecutive patients undergoing coronary angiography were included. Main clinical features and chemistry parameters were assessed at admission. 25(OH)D levels were measured by chemiluminescence immunoassay kit LIAISON® Vitamin D assay (Diasorin Inc, Stillwater, US). Hypovitaminosis D was defined for 25(OH)D<10ng/ml. A total of 3974 patients were included, of whom 29.4% had hypovitaminosis D. 25(OH)D deficiency was associated to age, female gender, diabetes mellitus, renal failure, previous percutaneous coronary intervention and smoke, acute presentation, severe coronary disease, higher glycemia and cholesterol and lower hemoglobin and ejection fraction (p<0.001), higher platelet count (p=0.004) and BMI (p=0.05). 25(OH)D significantly directly related with white blood cells count and the different components of leukocytes formula, Neutrophils-to-Lymphocytes Ratio, Monocytes-to-Lymphocytes Ratio and C-reactive protein, but not with lymphocytes levels. In fact, hypovitaminosis D predicted levels above the median for both Neutrophils-to-Lymphocytes Ratio (≥2.56; 57.3% vs. 47.6%; p<0.001; adjusted OR[95%CI]=1.28[1.07-1.52; p=0.007) and Monocytes -to-Lymphocytes Ratio (≥0.33; 59.1% vs. 49.8%; p<0.001; adjusted OR[95%CI]=1.3[1.1-1.54; p=0.002), results were confirmed in major subgroups of patients. The present study demonstrates that, among patients with cardiovascular disease, 25(OH)D deficiency is associated with a higher metabolic and clinical risk profile and with an elevation of cellular and humoral inflammatory parameters. Future dedicated studies should be, therefore, advocated in order to define whether 25(OH)D supplementation can modulate the mediators of the acute phase response and therefore potentially offer clinical and prognostic advantages on a broad spectrum of inflammatory disease.

Vitamin D levels condition the outcome benefits of renin-angiotensin system inhibitors (RASI) among patients undergoing percutaneous coronary intervention

BackgroundVitamin D deficiency is estimated as the most common medical condition worldwide, with severe implications on survival and on several inflammatory, immune-mediated and thrombotic disorders, and especially for cardiovascular disease.Recent studies have suggested that vitamin D could directly regulate the Renin-Angiotensin System (RAS) activity, therefore potentially interfering with the pharmacological effects of RAS Inhibitors (RASI), an issue that has seldom been explored. Therefore, the aim of the present study was to evaluate the prognostic impact of the use of RASI according to vitamin D levels among patients with coronary artery disease (CAD) undergoing percutaneous coronary intervention (PCI). MethodsConsecutive patients undergoing PCI were included. Main clinical features and chemistry parameters were assessed at admission. Vitamin D levels were measured by chemiluminescence immunoassay kit LIAISON® Vitamin D assay (Diasorin Inc). Severe deficiency was defined for 25(OH)D < 10 ng/mL. The primary study endpoint was defined as the occurrence of major cardiovascular events (MACE, a composite of death, recurrent Myocardial Infarction (MI) and target vessel revascularization) at the longest available follow-up. ResultsWe included a total of 705 patients, that were divided according to vitamin D tertiles (< 12.7 ng/mL; 12.7–21.59 ng/mL; ≥21.6 ng/mL) and use of RASI. RASI therapy was significantly associated to arterial hypertension, creatinine, lower 25(OH)D, use of statins, diuretics, ASA and ticagrelor across vitamin D tertiles. At a median follow-up of 996 [377–1552] days, MACE occurred in 174 (24.7 %) patients. Severe hypovitaminosis D was significantly associated with a higher rate of MACE (HR[95 %CI] = 0.75[0.62−0.91], p = 0.004). The use of RASI significantly lowered the rate of MACE in patients with lower vitamin D (I tertile: 41.3 % vs 25.9 %, adjusted HR[95 %CI] = 0.43[0.26−0.73], p = 0.002); whilst a non-significant effect was observed for II and III tertiles values (18.6 %vs 29.5 %, adjusted HR[95 %CI] = 1.16[0.57−2.34], p = 0.69, and 21.2 % vs 12.6 %, adjusted HR[95 %CI] = 1.1[0.46−2.62], p = 0.83) (p int = 0.04).A similar prognostic interaction for RASI and vitamin D was observed for cardiovascular mortality and MI (p int = 0.03). ConclusionAmong patients undergoing PCI, the use of RASI was associated with lower risk of MACE only among patients with lower levels of vitamin D. Future larger studies are certainly warranted in order to define the prognostic implications of vitamin D supplementation on the RAS system modulation, especially among patients treated with RASI.

Prognostic impact of Vitamin D deficiency in patients with coronary artery disease undergoing percutaneous coronary intervention

BackgroundWhether Vitamin D deficiency represents an independent predictor of mortality and major cardiovascular events or rather the mirror of a more advanced clinical condition with increased comorbidities is still debated. We aimed at assessing the impact of vitamin D levels on the long-term outcomes among patients with coronary artery disease (CAD) undergoing percutaneous coronary intervention. MethodsConsecutive patients from a single centre were included. Vitamin D levels were measured at admission by chemiluminescence immunoassay kit LIAISON® Vitamin D assay (Diasorin Inc). Severe deficiency was defined for 25(OH)D < 10 ng/ml. The primary study endpoint was overall mortality. Secondary endpoints were cardiovascular mortality, recurrent acute coronary syndrome or major cardiovascular events (a composite of death, recurrent MI and target vessel revascularization) at the longest available follow-up. ResultsWe included a total of 705 patients, that were divided according to vitamin D tertiles (<12.7; 12.7-21.59; ≥21.6 ng/ml). Lower levels of Vitamin D were associated with renal failure (p=0.03), more severe coronary disease (p=0.001), diabetes mellitus and previous CABG (p<0.001), lower ejection fraction (p=0.02), acute presentation (p=0.04), use of statins (p=0.02), diuretics, nitrates and clopidogrel (p<0.001) and RASI (p=0.008). An inverse association was documented with BMI, glycemia, total cholesterol (p<0.001), creatinine and WBC (p=0.001). At a median follow-up of 996.5 [377-1552] days, 3.8% of the patients died. Vitamin D deficiency was significantly associated with overall mortality (7.6% vs 2.9% vs 0.4%, adjusted HR[95%CI]=3.6[1.43-8.9], p=0.006), MACE (adjusted HR[95%CI]=1.32[1.07-1.63], p=0.01) and the composite of death and MI (adjusted HR[95%CI]=1.3[1.03-1.65], p=0.03). A similarly increased risk was confirmed for all major higher-risk subsets of patients, with no significant interaction according to age, gender, diabetes mellitus or chronic kidney disease. ConclusionAmong patients undergoing percutaneous coronary interventions, lower levels of vitamin D are associated with an over 3-fold increased risk of mortality and major cardiovascular events. Future larger studies are certainly warranted in order to define the prognostic implications of cholecalciferol supplementation among high-risk patients with established coronary artery disease.