Chemiluminescent Enzyme Immunoassay Research Articles

Anti-Human T-Cell Leukemia Virus Type 1 (HTLV-1) Antibody Assays in Cerebrospinal Fluid for the Diagnosis of HTLV-1-Associated Myelopathy/Tropical Spastic Paraparesis.

The anti-human T-cell leukemia virus type 1 (HTLV-1) antibody assay in common use has changed from the particle agglutination (PA) method to chemiluminescent immunoassay (CLIA) and chemiluminescent enzyme immunoassay (CLEIA). These assays were validated in serum but not in cerebrospinal fluid (CSF). However, anti-HTLV-1 antibody positivity in CSF is a requisite for diagnosing HTLV-1-associated myelopathy/tropical spastic paraparesis (HAM/TSP). We qualitatively compared the assays in CSF from 47 HAM/TSP patients diagnosed using PA, 15 HTLV-1 carriers (HCs), and 18 negative controls. In determining the positivity or negativity of CSF anti-HTLV-1 antibodies, we used serum cutoff points for CLIA and CLEIA because CSF cutoff points had not been decided. Truth table analysis revealed that the performance of CLIA was closer to that of PA and that CLEIA had low sensitivity. CSF antibodies from HAM/TSP patients were all positive by PA and CLIA but 83.0% positive by CLEIA. CSF antibodies from HCs were positive in 73.3%, 80.0%, and 6.7% by PA, CLIA, and CLEIA, respectively. Receiver operator characteristic curve analysis for CSF revealed that with the default cutoff point used for serum, CLIA and PA had comparable performances and CLEIA was less sensitive. The best performances of CLIA and CLEIA with adjusted cutoff points were 94.8% sensitivity and 95.5% specificity and 89.7% sensitivity and 95.5% specificity, respectively. We conclude that low-sensitivity CLEIA can underdiagnose HAM/TSP and that CLIA is a better alternative to PA in anti-HTLV-1 antibody assay for CSF with the current cutoff points.

Analytical validation of hepatitis B core-related antigen (HBcrAg) using dried blood spots (DBS).

Limited access to nucleic acid testing (NAT) to quantify HBV DNA levels, an essential tool to determine anti‐HBV treatment eligibility, represents a significant barrier to scale up HBV diagnostic services in resource‐limited countries. Hepatitis B core‐related antigen (HBcrAg) has the potential to become an affordable alternative because of its low cost (US$ <15/assay) and strong correlation with HBV DNA levels in treatment‐naïve patients. However, the current assay requires plasma or serum. To further facilitate its application to decentralized settings, we developed and evaluated a standardized procedure to quantify HBcrAg using dried blood spots as a tool to diagnose HBV‐infected people with high viraemia. We evaluated the following elution method optimized to quantify HBcrAg: suspension of a punched blood‐soaked disc (11 mm) of Whatman 903 Protein Saver Card in 450 µL of PBS 0.05% Tween 20, followed by an incubation for 4 h at room temperature and a centrifugation at 10,000 g for 10 minutes. 150 µL of DBS eluate was used to quantify HBcrAg using chemiluminescent enzyme immunoassay (LUMIPULSE® G600II, Fujirebio). The limit of detection of dried blood spot HBcrAg in relation with HBV DNA levels was 19,115 IU/mL across the five major HBV genotypes (A/B/C/D/E). A strong linear correlation was confirmed between dried blood spot HBcrAg and HBV DNA levels (r = 0.94, p < 0.0001) in samples with high viral loads (range: 3.7–7.0 log IU/mL). The coefficient of variation ranged between 4.0–11.2% for repeatability and 3.9–12.2% for reproducibility. Analytical specificity was 100% (95% CI: 83.9–100%) in HBV‐negative samples. Using our elution method, it may be possible to identify HBV‐infected patients with high viraemia who need antiviral therapy using dried blood spot and HBcrAg. A large‐scale clinical validation is warranted in resource‐limited countries.

Analytical and clinical comparison between two different chemiluminescent enzyme immunoassays for the measurement of C-peptide in serum

article: Analytical and clinical comparison between two different chemiluminescent enzyme immunoassays for the measurement of C-peptide in serum - Minerva Biotechnology and Biomolecular Research 2021 March;33(1):6-11 - Minerva Medica - Journals

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.

Immunochromatography and chemiluminescent enzyme immunoassay for COVID-19 diagnosis

IntroductionThe rapid and accurate detection of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is required to prevent the spread of COVID-19. This study evaluated the utility of two SARS-CoV-2 antigen detection methods. MethodsWe evaluated two types of antigen detection methods using immunochromatography (Espline) and quantitative chemiluminescent enzyme immunoassay (Lumipulse). RT-PCR was performed as a standard procedure for COVID-19 diagnosis. Lumipulse and RT-PCR were performed for all 486 nasopharyngeal swabs and 136 saliva samples, and the Espline test was performed for 271 nasopharyngeal swabs and 93 saliva samples. ResultsThe sensitivity and specificity of the Espline test were 10/11 and 260/260 (100%), respectively for the nasopharyngeal swabs and 3/9 and 84/84 (100%), respectively for the saliva samples. High sensitivities for both saliva (8/9) and nasopharyngeal swabs (22/24) were observed in the Lumipulse test. The specificities of the Lumipulse test for nasopharyngeal swabs and saliva samples were 460/462 (99.6%) and 123/127 (96.9%), respectively. ConclusionThe Espline test is not effective for saliva samples but is useful for simple and rapid COVID-19 tests using nasopharyngeal swabs because it does not require special devices. The Lumipulse test is a powerful high-throughput tool for COVID-19 diagnosis because it has high detection performance for nasopharyngeal swabs and saliva samples.

A novel rapid immunoassay of serum type IV collagen 7S for the diagnosis of fibrosis stage of nonalcoholic fatty liver diseases.

Type IV collagen 7S (T4C7S) is a valuable biomarker for detecting liver fibrosis in patients with nonalcoholic fatty liver disease (NAFLD). The conventional T4C7S measurement via radioimmunoassay (T4C7S RIA) has shortcomings of radioisotope usage and longer assay periods. We compared T4C7S RIA with a newly developed, fast T4C7S chemiluminescent enzyme immunoassay (T4C7S CLEIA) and examined the diagnostic accuracies of and correlation between the two techniques. We evaluated 170 biopsy-confirmed patients with NAFLD. T4C7S was measured via both T4C7S RIA and T4C7S CLEIA. The correlation between T4C7S RIA and T4C7S CLEIA was analyzed in 305 total serum samples via exploratory research and 47 validation samples. The diagnostic accuracies of T4C7S CLEIA and T4C7S RIA were compared in the sera of patients with NAFLD and test samples. Sera T4C7S levels of T4C7S CLEIA and T4C7S RIA significantly correlated in patients' samples via exploratory (r=0.914, P=0.000) and validation (r=0.929, P=0.000) research. At a 10% coefficient, T4C7S CLEIA concentration was 0.26ng/ml in the serum samples, indicating high accuracy at even low concentrations. T4C7S CLEIA revealed distinct changes between each stage and high sensitivity in detecting the F2 stage, indicating a higher sensitivity in detecting low fibrosis stages than T4C7S RIA in patients with NAFLD. The T4C7S CLEIA correlated well with the T4C7S RIA. Favorably, the T4C7S CLEIA has a higher sensitivity and rapid measurement time and requires a small sample volume; thus, it is a promising and popular biomarker for fibrosis stage diagnosis in NAFLD.

Novel chemiluminescent immunoassay to measure plasma aldosterone and plasma active renin concentrations for the diagnosis of primary aldosteronism

Determination of plasma aldosterone concentrations (PAC) and plasma active renin concentrations (ARC) is essential for the diagnosis of primary aldosteronism (PA). In Japan, although PAC and ARC are measured by radioimmunoassay and immunoradiometric assay, respectively, non-radioisotopic methods with better detection sensitivity, measurement accuracy, and technical simplicity are needed. We developed two-site sandwich chemiluminescent enzyme immunoassays (CLEIAs) to measure both PAC and ARC using monoclonal antibodies immobilized onto ferrite particles. The results of both assays are obtained simultaneously from a single plasma sample within 30 min using a fully automated system. The novel CLEIAs were validated using plasma samples from patients with PA (n = 52) and essential hypertension (n = 23). The PAC determined by the CLEIA was significantly correlated with that measured by liquid chromatography/mass spectrometry or conventional radioimmunoassay. The ARC determined by the CLEIA was significantly correlated with that measured by immunoradiometric assay. The limits of detection of the CLEIAs for PAC and ARC were 0.1 ng/dl and 0.04 pg/ml, respectively, which were better than those of conventional methods (PAC: 2.5 ng/dl; ARC: 5 pg/ml). The PAC and PAC/ARC ratio (ARR) were significantly higher, and the ARC significantly lower, in patients with PA than in those with essential hypertension. An ARR cut-off of 1.31 ng/dl per pg/ml showed a sensitivity of 96.2% and specificity of 78.3% for PA screening. The newly developed CLEIAs for measuring PAC and ARC could provide a clinically powerful alternative to conventional methods used for hypertension screening in clinical practice.

Antibody seroprevalence in the epicenter Wuhan, Hubei, and six selected provinces after containment of the first epidemic wave of COVID-19 in China

BackgroundChina implemented containment measures to stop SARS-CoV-2 transmission in response to the COVID-19 epidemic. After the first epidemic wave, we conducted population-based serological surveys to determine extent of infection, risk factors for infection, and neutralization antibody levels to assess the real infections in the random sampled population. MethodsWe used a multistage, stratified cluster random sampling strategy to conduct serological surveys in three areas - Wuhan, Hubei Province outside Wuhan, and six provinces selected on COVID-19 incidence and containment strategy. Participants were consenting individuals >1 year old who resided in the survey area >14 days during the epidemic. Provinces screened sera for SARS-CoV-2-specific IgM, IgG, and total antibody by two lateral flow immunoassays and one magnetic chemiluminescence enzyme immunoassay; positive samples were verified by micro-neutralization assay. FindingsWe enrolled 34,857 participants (overall response rate, 92%); 427 were positive by micro-neutralization assay. Wuhan had the highest weighted seroprevalence (4•43%, 95% confidence interval [95%CI]=3•48%-5•62%), followed by Hubei-ex-Wuhan (0•44%, 95%CI=0•26%-0•76%), and the other provinces (<0•1%). Living in Wuhan (adjusted odds ratio aOR=13•70, 95%CI= 7•91–23•75), contact with COVID-19 patients (aOR=7•35, 95%CI=5•05–10•69), and age over 40 (aOR=1•36, 95%CI=1•07–1•72) were significantly associated with SARS-CoV-2 infection. Among seropositives, 101 (24%) reported symptoms and had higher geometric mean neutralizing antibody titers than among the 326 (76%) without symptoms (30±2•4 vs 15±2•1, p<0•001). InterpretationThe low overall extent of infection and steep gradient of seropositivity from Wuhan to the outer provinces provide evidence supporting the success of containment of the first wave of COVID-19 in China. SARS-CoV-2 infection was largely asymptomatic, emphasizing the importance of active case finding and physical distancing. Virtually the entire population of China remains susceptible to SARS-CoV-2; vaccination will be needed for long-term protection. FundingThis study was supported by the Ministry of Science and Technology (2020YFC0846900) and the National Natural Science Foundation of China (82041026, 82041027, 82041028, 82041029, 82041030, 82041032, 82041033).

Re-evaluation of potential predictors of calretinin and mesothelin in a population-based cohort study using assays for the routine application in clinical medicine

ObjectivesCalretinin and mesothelin are molecular markers for the detection of malignant mesothelioma at early stages. Our objective was the re-evaluation of factors influencing calretinin and mesothelin concentrations in plasma of...

Serum KL-6 Could Represent a Reliable Indicator of Unfavourable Outcome in Patients with COVID-19 Pneumonia.

KL-6 is a sialoglycoprotein antigen which proved elevated in the serum of patients with different interstitial lung diseases, especially in those with a poorer outcome. Given that interstitial pneumonia is the most common presentation of SARS-CoV2 infection, we evaluated the prognostic role of KL-6 in patients with COVID-19 pneumonia. Patients with COVID-19 pneumonia were prospectively enrolled. Blood samples were collected at the time of enrolment (TOE) and on day 7 (T1). Serum KL-6 concentrations were measured by chemiluminescence enzyme immunoassay using a KL-6 antibody kit (LUMIPULSE G1200, Fujirebio) and the cut-off value was set at >1000 U/mL. Fifteen out of 34 enrolled patients (44.1%) died. Patients with unfavourable outcome showed significantly lower P/F ratio and higher IL-6 values and plasmatic concentrations of KL-6 at TOE compared with those who survived (median KL-6: 1188 U/mL vs. 260 U/mL, p < 0.001). KL-6 > 1000 U/mL resulted independently associated with death (aOR: 11.29, p < 0.05) with a positive predictive value of 83.3%. Our results suggest that KL-6 is a reliable indicator of pulmonary function and unfavourable outcome in patients with COVID-19 pneumonia. A KL-6 value > 1000 U/mL resulted independently associated with death and showed good accuracy in predicting a poorer outcome. KL-6 may thus represent a quick, inexpensive, and sensitive parameter to stratify the risk of severe respiratory failure and death.

Prognostic bioindicators in severe COVID-19 patients

BackgroundSevere acute respiratory syndrome caused by novel coronavirus 2 (SARS-CoV-2) emerged in Wuhan (China) in December 2019. Here we evaluated a panel of biomarkers to phenotype patients and to define the role of immuno-inflammatory mediators as biomarkers of severity. Materials and methodsSerum samples were obtained from 24 COVID-19 patients on admission to hospital, before any treatment or infusion of intravenous steroids or invasive ventilation. KL-6 IL-6 and C-peptide were measured by chemiluminescent enzyme immunoassay. IL-6 assay was validated for accuracy and precision. The validity of variables used to distinguish severe from mild-to-moderate patients was assessed by areas under curves (AUC) of the receiver operating characteristic (ROC) and logistic regression was performed to combine parameters of the two groups. ResultsIn the severe group, IL-6, CRP and KL-6 concentrations were significantly higher than in mild-to-moderate patients. KL-6, IL-6 and CRP concentrations were directly correlated with each other. ROC curve analysis of the logistic regression model including IL-6, KL-6 and CRP showed the best performance with an AUC of 0.95. ConclusionsBesides corroborating previous reports of over-expression of IL-6 in severe COVID-19 patients requiring mechanical ventilation, analytical determination of other mediators showed that IL-6 concentrations were correlated with those of KL-6 and CRP. The combination of these three prognostic bioindicators made it possible to distinguish severe COVID-19 patients with poor prognosis from mild-to-moderate patients.

Logistic Advantage of Two-Step Screening Strategy for SARS-CoV-2

Background: Airport quarantine is required to reduce the risk of entry of travelers infected with severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). However, it is challenging for both high accuracy and rapid turn-around time to coexist in testing; polymerase chain reaction (PCR) is time-consuming with high accuracy, while antigen testing is rapid with less accuracy. Methods: 88,924 (93.2%) of 95,457 arrivals at three international airports in Japan were tested for SARS-CoV-2 using self-collected saliva by a screening strategy with initial chemiluminescent enzyme immunoassay (CLEIA) followed by confirmatory nucleic acid amplification tests (NAAT) only for intermediate range antigen concentrations. Findings: 254 (0.27%) persons were found to be SARS-CoV-2 antigen positive (≥ 4.0 pg/mL) by CLEIA. NAAT was required for confirmatory testing in 513 (0.54%) persons with intermediate antigen concentrations (0.67-4.0 pg/mL) whereby the virus was detected in 34 (6.6%) persons. This two-step strategy dramatically reduced the utilization of NAAT to approximately one out of every 200 test subjects. Estimated performance of this strategy did not show significant increase in false negatives as compared to performing NAAT in all subjects. Further reduction in imported cases may be achieved by post-screening quarantine. Interpretation: Point of care testing by quantitative CLEIA using self-collected saliva is less labor-intensive and yields results rapidly, thus suitable as an initial screening test. Reserving NAAT for CLEIA indeterminate cases may prevent compromising accuracy while significantly improving the logistics of administering mass-screening at large venues.Funding Statement: Health, Labour and Welfare Policy Research Grants 20HA2002.Declaration of Interests: We declare no competing interests.Ethics Approval Statement: This study was approved by the Institutional Ethics Board (Hokkaido University Hospital Division of Clinical Research Administration Number: 020-0116) and anonymously processed data were provided by the quarantine stations.

Development of an Automated Chemiluminescence Assay System for Quantitative Measurement of Multiple Anti-SARS-CoV-2 Antibodies.

ObjectivesSerological tests for COVID-19 have been instrumental in studying the epidemiology of the disease. However, the performance of the currently available tests is plagued by the problem of variability. We have developed a high-throughput serological test capable of simultaneously detecting total immunoglobulins (Ig) and immunoglobulin G (IgG) against nucleocapsid protein (NP) and spike protein (SP) and report its performance in detecting COVID-19 in clinical samples.MethodsWe designed and prepared reagents for measuring NP-IgG, NP-Total Ig, SP-IgG, and SP-Total Ig (using N-terminally truncated NP (ΔN-NP) or receptor-binding domain (RBD) antigen) dedicated automated chemiluminescent enzyme immunoassay analyzer AIA-CL1200. After determining the basal thresholds based on 17 sera obtained from confirmed COVID-19 patients and 600 negative sera, the clinical validity of the assay was evaluated using independent 202 positive samples and 1,000 negative samples from healthy donors.ResultsAll of the four test parameters showed 100% specificity individually (1,000/1,000; 95%CI, 99.63–100). The sensitivity of the assay increased proportionally to the elapsed time from symptoms onset, and all the tests achieved 100% sensitivity (153/153; 95%CI, 97.63–100) after 13 days from symptoms onset. NP-Total Ig was the earliest to attain maximal sensitivity among the other antibodies tested.ConclusionOur newly developed serological testing exhibited 100% sensitivity and specificity after 13 days from symptoms onset. Hence, it could be used as a reliable method for accurate detection of COVID-19 patients and to evaluate seroprevalence and possibly for surrogate assessment of herd immunity.

Seroepidemiological Survey of the Antibody for Severe Acute Respiratory Syndrome Coronavirus 2 with Neutralizing Activity at Hospitals: A Cross-sectional Study in Hyogo Prefecture, Japan.

Introduction: The coronavirus disease 2019 (COVID-19) pandemic is spreading rapidly all over the world. The Japanese government lifted the state of emergency, announced in April 2020, on May 25, but there are still sporadic clusters. Asymptomatic patients who can transmit severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) cause some of these clusters. It is thus urgent to investigate the seroprevalence of antibodies against SARS-CoV-2 and their neutralizing activity. We conducted a cross-sectional study of >10,000 samples at hospitals in Hyogo Prefecture, Japan. Methods: Between August 6 and October 1, 2020, we collected samples of residual blood from the patients who visited or were admitted to five hospitals and a foundation in Hyogo. We tested the samples for antibodies against SARS-CoV-2 by electrochemiluminescence immunoassay (ECLIA) and chemiluminescent enzyme immunoassay (CLEIA). Sera that were positive by ECLIA or CLEIA were analyzed by an immunochromatographic (IC) test and neutralizing activity assay. Results: We tested 10,377 samples from patients aged between 0 and 99 years old; 27 cases (0.26%) were positive on the ECLIA, and 51 cases (0.49%) were positive on CLEIA. In the 14 cases that tested positive on both ECLIA and CLEIA, the positive rates on the IC test and for neutralizing activity were high (85% and 92%, respectively). In 50 cases (0.48%) that were positive by either ECLIA or CLEIA, the corresponding rates were low (20% and 6%, respectively). The positive rate of neutralizing antibody was 0.15%. Conclusions: These results indicate that most Hyogo Prefecture residents still do not have antibodies and should avoid the risk of incurring a SARS-CoV-2 infection. Two or more antibody tests should be required for seroepidemiological studies of the antibody for SARS-CoV-2, and a neutralizing activity assay is also essential.

SARS-CoV-2 Seroprevalence and Accuracy Diagnostic Evaluation During a Covid-19 Outbreak in a Major Penitentiary Complex in Brazil, June-July 2020: A Population-Based Study

Summary: The pandemic of COVID-19 has presented as a threat to prison systems worldwide, as overcrowding, limited possibility of social distancing and sanitary restrictions favouring viral spread. SARS-CoV-2 seroprevalence studies in prison settings are important tools to understand transmission and to develop control measures. We aimed to estimate the overall SARS-CoV-2 seroprevalence and evaluated the accuracy of an antibody rapid test compared to a reference serological assay during a COVID-19 outbreak in a prison complex housing over 13,000 inmates in Brasília. Methods: We conducted a repeated serosurvey of IgM/IgG SARS-CoV-2 antibodies among inmates between June 17-July 1 (baseline) and July 8-22, 2020 (21-days follow-up). We obtained a randomized, stratified representative sample of each prison unit. Inmates who accepted to be part of the study were interviewed through a standardised questionnaire covering sociodemographic and clinical characteristics. Blood samples were collected from each participant for the evaluation of SARS-CoV-2 antibodies with a lateral-flow immunochromatographic assay (LFIA). Inmates with negative results in the first serosurvey were interviewed and tested for a second time to evaluate recent symptoms history and serological status 21 days later. Samples were also retested using a chemiluminescence enzyme immunoassay (CLIA) to compare SARS-CoV-2 seroprevalence and 21-days incidence, as well as to estimate the overall infection fatality rate (IFR) and determine the diagnostic accuracy of the LFIA test.Findings: We identified 485 eligible individuals and enrolled 460 participants (94.8%), distributed across four prison units. Baseline and 21-days follow-up seroprevalence were estimated at 52.0% (95%CI 44.9-59.0) and 56.7% (95%CI 48.2-65.3) with the LFIA test; and 80.7% (95%CI 74.1-87.3) and 81.1% (95%CI 74.4-87.8) with the CLIA test, with an overall IFR of 0.02%. There were 78.2% (95%CI 66.7-89.7) symptomatic inmates among the positive cases, and the main symptoms included changes in taste or smell, headache, and fever. Sensitivity and specificity of LFIA test were estimated at 43.4% and 83.3% for IgM; 46.5% and 91.5% for IgG; and 59.1% and 77.3% for combined tests. There was a fair agreement between LFIA and CLIA results (Kappa score 0.23).Interpretation: We found rapid viral spread and high seroprevalence of anti-SARS-CoV-2 antibodies within the prison complex. The occurrence of asymptomatic infection highlights the importance of periodic mass testing in addition to case-finding of symptomatic inmates, however the field performance of LFIA tests should be validated. We recommend that vaccination strategies consider the inclusion of persons deprived of liberty and prison staff in priority groups.Funding: EN, GASR, RH, WNA acknowledge the Brazilian Ministry of Education (MEC) for support the COVID-19 response by University of Brasília (#23106.028855/2020-74). RH, WMR and WNA acknowledge the Federal District Research Foundation (FAP-DF) for grants that supported this research (# 00193-00000495/2020-72). GASR, JHC, WMR, WNA acknowledge the National Research Council (CNPq) for grants that supported this research (# 402957/2020-2)Declaration of Interest: None to declare. Ethical Approval: The study was part of an epidemiological investigation conducted by the Brazilian Field Epidemiology Training Program/Ministry of Health as a response to a public health emergency, and it is supported by the national law 8.080/1990. Moreover, all participants provided informed consent and ethics approval was obtained under the CONEP protocol number 37007220.1.0000.0008.

Role of Serum Total Testosterone and High Sensitivity C-Reactive Protein Levels in Type 2 Diabetic Males

Introduction: Diabetes Mellitus (DM) is a chronic metabolic disorder associated with low level of Total Testosterone (TT) and Hypogonadism. Estimation of High sensitivity C-Reactive Protein (Hs-CRP) is useful for assessing the cardiovascular risk. Aim: To estimate serum TT and Hs-CRP levels in Diabetic patients and hence, the association of Diabetes with Erectile Dysfunction (ED) and Cardiovascular risk. Materials and Methods: This was a case-control study with a case-control design, conducted at a tertiary care centre on 100 subjects,60 DM males as cases and 40 healthy male volunteers of same age, as controls, from June 2016 to May 2017. All the relevant demographic and clinical details i.e. age, height, weight, body mass index (BMI), Systolic and Diastolic Blood Pressure were noted for all subjects. All relevant tests i.e. serum TT by Electrochemiluminescence immunoassay analyser Enzyme Chemiluminescence Immunoassay (ECLIA) method, serum Hs-CRP by immunoturbidimetry method, fasting and post-prandial glucose measurements, HbA1c were all done and results were tabulated and analysed. Chi-square test was used to calculate p-value. Analysis of Variance (ANOVA) descriptive analysis was used to find out the mean, standard deviation, standard error and 95% confidence interval. Results: The mean of HbA1c among the case group was 8.185% and 5.004% among the controls. The mean value of TT among cases was 3.891 ng/dL, which was significantly reduced (p-value &lt;0.005) as compared to healthy controls i.e. 5.339 ng/dL. Low level of TT (below 3.5 ng/dL) was seen in 45% of the cases among the diabetic study population which was statistically significant (p-value &lt;0.0001). Seventy five percent of diabetics had erectile dysfunction. The mean value of Hs-CRP among diabetic population was 3.875 and controls were 1.457 mg/L (p-value 0.001). Based on Hs-CRP values, the subjects were sub-categorised into low, intermediate and high risk for CVD. A 31.7% of diabetics were in the intermediate group and 68.3% were in the high risk group for CVD. Conclusion: There was significant reduction in TT in Type 2 Diabetic men. Patients with low TT had increased Hs-CRP value among which 75% falling under high risk category and 25% under intermediate risk category. All the cases had serum Hs- CRP value of either intermediate or high risk category. Low level of TT was also associated with ED and PME.

Association of circulating soluble lectin-like oxidized low-density lipoprotein receptor-1 with inflammatory markers and urinary albumin excretion in patients with type 2 diabetes.

Objectives:The main purpose of the study was to study the association between circulating soluble lectin-like oxidized low-density lipoprotein receptor-1 levels and various markers, including inflammatory markers such as high-sensitivity C-reactive protein and fibrinogen, serum lipids, and renal function, in patients with poorly controlled type 2 diabetes.Methods:The subjects were 70 patients (men 45, women 25) who were hospitalized for treatment of poor glycemic control. Plasma soluble lectin-like oxidized low-density lipoprotein receptor-1 levels were assayed using a sandwich chemiluminescence enzyme immunoassay.Results:Circulating soluble lectin-like oxidized low-density lipoprotein receptor-1 was significantly positively correlated with lectin-like oxidized low-density lipoprotein-1 ligands containing apolipoprotein B, reflecting modified low-density lipoprotein, and with inflammatory markers such as high-sensitivity C-reactive protein and fibrinogen. In addition, there was a significant positive correlation between soluble lectin-like oxidized low-density lipoprotein receptor-1 and urinary albumin excretion.Conclusions:Soluble lectin-like oxidized low-density lipoprotein receptor-1 may serve as a marker reflecting the degrees of inflammation and albuminuria in patients with poorly controlled type 2 diabetes.

Evaluation of the clinical utility of disease-specific antinuclear antibody screening tests using chemiluminescence enzyme immunoassay for routine antinuclear antibody testing

Evaluation of the clinical utility of disease-specific antinuclear antibody screening tests using chemiluminescence enzyme immunoassay for routine antinuclear antibody testing

Immunogenicity and Safety of Inactivated SARS-CoV-2 Vaccine in People Living with HIV: A Cross-Sectional Study

Background: To analyze the efficacy and safety of SARS-CoV-2 inactivated vaccine in people living with HIV (PLWH).Methods: A total of 143 PLWH were included in the study. All patients were confirmed with HIV-1 infection. We also enrolled 50 healthy individuals vaccinated with two doses of SARS-CoV-2 vaccine as controls. A commercially available magnetic chemiluminescence enzyme immunoassay kit was used to detected serum IgG and IgM against SARS-CoV-2.Findings: Serum levels of SARS-CoV-2-specific IgG were significantly higher in the control group than in the PLWH group (P=0.001). Overall, 76% of individuals in the control group achieved IgG seroconversion after vaccination compared with 58% in the PLWH group (P=0.024). The time after vaccination in IgG seronegative PLWH was significantly longer compared with PLWH with IgG seropositive (43.38 ± 34.96 vs 30.27 ± 20.12 days, P=0.005). In PLWH with IgG seropositivity, CD4+ T cell counts before antiretroviral therapy (ART) (P=0.015) and at IgG detection (PInterpretation: The inactivated COVID-19 vaccine appears to be safe with good immunogenicity in PLWH.Funding: This study was supported by Clinical Research Startup Program of Southern Medical University by High-level University Construction Funding of Guangdong Provincial Department of Education (No. LC2016PY003).Declaration of Interest: None of the authors have competing interests to disclose.Ethical Approval: The study was performed in accordance with the Declaration of Helsinki and was approved by the Institutional Ethics Committee of Nanfang Hospital (NFEC-2021-178).

Concentration of high-sensitivity cardiac troponin I in the oral fluid in patients with acute myocardial infarction: a pilot study

Aim . To assess the potential of using oral fluid as a non-invasive diagnostic material in patients with myocardial infarction (MI). Material and methods . The pilot, single-center, prospective study included 47 patients with documented MI, among whom there were 33 men (71%) and 14 women (29%) (mean age, 61,72±12,09 years). All patients successfully treated with reperfusion therapy. The control group consisted of 15 people in whom MI was not confirmed. The concentration of high-sensitivity cardiac tro -ponin I ( hs-cTnI) in blood and oral fluid was determined using chemilumines -cence enzyme immunoassay (CLEIA) on a PATHFAST analyzer (LSI Medience Corporation). Medium sensitivity cardiac troponin I (ms-cTnI) was determined in blood using an Access 2 immunoassay system analyzer (Beckman Coulter, USA). Levels of total bilirubin, creatinine, glucose, rheumatoid factor, alkaline phosphatase and other biochemical parameters were determined on a Furuno CA-400 analyzer (Japan). Results . The levels of hs-cTnI in patients with MI were significantly higher than in healthy patients both in blood (8,73±1,17 ng/ml vs 0,012±0,03 ng/ml, p&lt;0,001) and oral fluid (0,41±0,11 ng/ml vs 0,004±0,001 ng/ml, p&lt;0,001). In patients with-AMI, there was a moderate correlation between the concentration of hs-cTnI in the serum and the oral fluid (r=0,319; p&lt;0,05). The serum level of hs-cTnI in patients with Q-wave (n=33) and non-Q-wave (n=14) MI was 10,11±1,53 ng/ ml vs 5,48±1,29 ng/ml, respectively (p=0,025). The oral fluid concentration of hs-cTnI in patients with W-wave (n=33) and non-Q-wave (n=14) MI was 0,42±0,14 ng/ml vs 0,40±0,16 ng/ml, respectively (p=0,925). The serum level of hs-cTnI in anterior MI (n=19) was 8,92±2,06 ng/ml vs 8,91±1,81 ng/ml in posterior one (n=23) (p=0,997). The concentration of hs-cTnI in the oral fluid was 0,21±0,06 ng/ml vs 0,57±0,21 ng/ml, respectively (p=0,107). The oral fluid concentrations of hs-TnI in patients with MI using conventional plastic tubes (n=26) and special Sarstedt microtubes (n=21) were 0,56±0,19 ng/ml and 0,22±0,10 ng/ml, respectively (p=0,12). Conclusion. This pilot study has proven the possibility of detecting hs-cTnI in the oral fluid of patients with MI. There was a moderate correlation between the level of hs-cTnI in blood serum and oral fluid. Further research is needed to determine the hs-cTnI reference values in the oral fluid of patients with MI.