Roche Diagnostics GmbH Research Articles

B-320 Understanding CA 72-4: Exploring False Positives in Clinical Context

Abstract Background CA 72-4, initially identified in 1981 as an antigen reactive to murine antibodies, is a glycoprotein expressed on the cell surface of certain carcinoma cells. Its production is elevated in various cancers, including ovarian, pancreatic, colorectal, and gastric cancer. However, CA 72-4 serum concentration may rise in non-cancerous conditions, leading to potential misdiagnosis and a low positive predictive value (PPV), particularly in asymptomatic patients. Our aim in this study was to describe CA 72-4 false positive values and assess the pathological conditions and pharmacological treatments associated with this. We also intended to compare other tumour markers status in false and true positive results of CA 72-4. Methods A retrospective analysis form pseudo-anonymized database was conducted using data from 1290 laboratory analysis from different patients taking a blood test at our laboratory hospital from 2017 to 2019. CA 72-4 and other tumour markers were analysed by ECLIA (Roche Diagnostics Gmbh. Mannheim, Germany). Results Among 1290 CA 72-4 laboratory tests in different patients, 204 were considered positive because they were above the reference interval (<7 U/mL). Within this subgroup, 44 of the 204 were clearly classified as false positives, as patients were not diagnosed with any form of neoplastic process. 45% of cases were between 7-14 U/mL (group 1), 30% were between 14-28 U/mL (group 2), and finally 25% of patients were above 28 U/mL ranging from 35.1 to 240.4 U/mL (group 3). Interestingly, CA19-9, CA 125 and CEA were measured in 37 of the false positive cases and it was found that 59.5% had at least another tumour marker above the reference range. In 103 true positive cases it was found that 88% of patients had at least one tumour marker above the normal range. Patients exhibiting false positive results for CA 72-4 demonstrated a wide spectrum of pathological conditions, with pulmonary disease, inflammation, infection, and heart and kidney disease emerging as the most prevalent. Group 1 patients most common underlying pathologies were pulmonary disease in 25%, heart disease in 25% and either viral or bacterial infection in 60% of patients. Group 2 patients most common pathologies were heart disease in 42%, pulmonary disease in 33% and infection in 25% of patients. In group 3 patients, renal disease was present in 45% of patients, heart disease was found in 45% of patients and inflammation/sepsis was found in 36% of patients. When looking into pharmacological treatment, 86.3% of patients were taking at least one of previously described treatments increasing CA 72-4 in serum; proton pump inhibitors, corticosteroids, antiplatelet drugs and colchicine. Drug treatment proportion was very similar across the different groups. Conclusions CA72-4 false positive cases are sometimes in conjunction with other false positive tumour markers but not in the same proportion as true positives. In addition, most false positive patients can be explained by pharmacological treatments. This research may help in the understanding and identification of potential factors influencing tumor marker levels and thus contribute to the interpretation of results.

THE ASSOCIATION BETWEEN HEPATITIS B VIRUS INFECTION WITH ABO AND RHD BLOOD GROUPS

Background: ABO and Rh blood types are associated with a higher risk of disease and are essential for clinical practice and blood transfusion safety. The findings of earlier research on the relationship between blood types and HBV infection are still up for debate. There is no study conducted on the study population to reveal the relationship between ABO and RhD blood groups and infection with hepatitis B virus. Aims: This study aimed to identify the relationship between blood types and hepatitis B virus infection in blood donors at Jiblah University Hospital in Jiblah town, Yemen Methods: A hospital-based cross-sectional study was conducted. Venous blood samples were drawn from 120 blood donors and tested for hepatitis B core antigen (anti-HBc) using commercial kits (Roche Diagnostics GmbH, United Kingdom). ABO and Rh blood groups were determined by the cell pooling method. The results were analyzed using SPSS v.20. Results: This study reported that the most frequent blood types were O positive and A positive 57 (47.5%) and 28 (23.3%) respectively, followed by B positive 14 (11.67%) and O negative 13 (10.83%). The overall phenotypic frequencies of ABO and RhD blood groups were O+>A+>B+>O->A->AB+>B->AB-. The prevalence of hepatitis B virus was 12 (10%). No statistical association was detected between ABO and RhD blood groups and hepatitis B infection. Conclusions: High prevalence rates of hepatitis B virus infection among blood donors were identified in this study. There was no statistical association between ABO and RhD blood groups and hepatitis B infection. Peer Review History: Received 25 March 2024; Revised 3 May 2024; Accepted 22 June; Available online 15 July 2024 Academic Editor: Prof. Cyprian Ogbonna ONYEJI, Obafemi Awolowo University, Ile-Ife, Nigeria, conyeji@oauife.edu.ng Average Peer review marks at initial stage: 6.0/10 Average Peer review marks at publication stage: 7.0/10 Reviewers: Dr. Leyla Açık, Gazi University, Ankara, Turkey, leylaacik@gmail.com Prof. Hassan A.H. Al-Shamahy, Sana'a University, Yemen, shmahe@yemen.net.ye

Impact of COVID-19 on pro- and antiatherogenic lipoproteins (cross-sectional population study)

Aim. The aim of this cross-sectional retrospective study was to study the effect of SARS-CoV-2 S1/S2 infection on population lipid parameters, which are leading risk factors for the development and progression of atherosclerosis, which can be significantly distorted in systemic inflammation and, in particular, during respiratory viral infections.Material and methods. We analyzed anonymized results of one-time, one-year studies of complete lipid profiles and related laboratory parameters performed in the Helix Laboratory Service from February 1, 2015 to December 30, 2020 in 238541 males and 384437 females aged from 22 to 83 years in 334 populated areas of the European Russia using Roche Cobas C502, C702 (Roche Diagnostics GmbH, Mannheim, Germany), LIAISON XL (DiaSorin S.p.A, Italy) analyzers.Statistical analysis included methods of descriptive statistics, distribution analysis, sample comparisons, and search for dependencies.Results. A dramatic change in the magnitude and nature of seasonal population fluctuations in low-density lipoprotein cholesterol and high-density lipoprotein cholesterol (HDL-C) during the COVID-19 spread has been identified.COVID-19 differentially affects the relationship between high-sensitivity C-reactive protein (hsCRP) and atherogenic and antiatherogenic lipoproteins. These relationships have sex differences, are nonlinear, and in relation to HDL-C are associated with the level of specific anti-SARS-CoV-2 S1/S2 antibodies.Up to a hsCRP level of 2,5 mg/l, there is a significant increase in population levels of low-density lipoprotein cholesterol with a correlation coefficient of 0,14 for women (p<0,001) and 0,10 for men (p<0,001). At hsCRP levels >2,5 mg/l, the trend reverses. At the same time, HDL-C levels sharply decrease with a negative correlation of -0,23 (p<0,001) in women and -0,22 (p<0,001) in men with hsCRP values <2,5 mg/l, followed by a less pronounced decline.Conclusion. The study results may be useful for optimal prevention development and adequate assessment of atherogenic dyslipidemia treatment effectiveness in patients after COVID-19.

Plasma D-Dimer Value Corrected with Some Physiological and Inflammatory Markers (C-Reactive Protein and Ferritin) in Iraqi Patients with COVID-19 Infection

Since the beginning of the COVID-19 pandemic it has been observed that patients have elevated plasma levels of D-dimer and some inflammatory markers (ferritin, interleukine 6, C-reactive protein (CRP) or fibrinogen). Some studies point to the existence of a certain correlation between those markers and D-dimer. CRP is a protein discovered in the 1930s by Tillett and Francis and is an acute phase reactant. It is a pentameric protein which is synthesized by the liver under the action of cytokine interleukin 6 (IL-6). D-dimers are multiple peptide fragments produced as a result of degradation of crosslinked fibrin, mediated by plasmin. A total of 60 patients were recruited and categorized into :1- group1 (controls), 2- group 2 (COVID-19 patients). 5 ml of blood was obtained from each patient by vein puncture, using 5 ml disposable syringes, then centrifuged at 3000 rpm for 10 minutes to collect the serum. D- dimer, C-reactive protein were measured by using (Roche Diagnostics GmbH, Mannheim, Germany). At the same time, the Ferritin was assessed by using a miniVIDAS analyzer for the fluorescent enzymatic detection of β2-microglobulin (β2M) using the technique. Enzyme Linked Fluorescent Assay (ELFA) (BioMerieux). Our results showed that there was a non- significant difference in the P-values between control and patients males and females. The Mean ± SE of age in control group was 45.90 ± 3.34, while the Mean ± SE of age in patients was 45.35 ± 2.52. There was a non- significant difference between the two groups, the Mean ± SE of CRP in control group was 5.04 ±0.81, while the Mean ± SE of CRP in COVID-19 patients was 37.16 ±3.24, there was a highly Significant differences between them (P≤0.01). The CRP of COVID-19 patients were compared with those of control patients, the results shows significant increased CRP in covid-19 patients group as a compression with the control group, the findings of the study is similar to Jacob Lentner, etal findings. In response to infections, the liver synthesizes significant quantities of acute-phase proteins (APPs), such as CRP. This acute inflammatory protein is a highly sensitive biomarker for inflammation, tissue damage, and infection. It has been shown that CRP levels are correlated with levels of inflammation. CRP levels can promote phagocytosis and activate the complement system. In other words, CRP binds to microorganisms and promotes their removal through phagocytosis. D-dimers are one of the fragments produced when plasmin cleaves fibrin to break down clots. Our study showed that the serum D-dimer concentrations in patients significantly higher than those in control group which is similar to Mamta Soni, etal, 2020 findings. Elevated D-dimer levels have emerged as a consistent finding in severely ill COVID-19 patients, Multiple studies have identified an association between higher D-dimer levels and an increased risk of mortality in the COVID-19 patient population. Ferritin is an iron-storing protein; its serum level reflects the normal iron level and helps the diagnosis of iron deficiency anemia. Circulation ferritin level increases during viral infections and can be a marker of viral replication. Our study showed a significant increase in ferritin level in covid-19 patients group compared to control group, which consider similar to [] results. Although the exact cause for elevated ferritin in COVID-19 infection is unknown, it could be influenced by cytokine release or cellular damage that results in the leakage of intracellular ferritin. It has been previously shown that ferritin is a direct indicator of cellular damage suggestive of an association between organ damage and ferritin production. This could later cause cell death, known as ferroptosis. It is suggested that inflammation associated with sepsis could alter iron metabolism and deficiency to facilitate the immune system, which could be an early sign of COVID.

Performance Evaluation of Three Antibody Binding Assays, a Neutralizing Antibody Assay, and an Interferon-Gamma Release Assay for SARS-CoV-2 According to Vaccine Type in Vaccinated Group.

We evaluated the performance of SARS-CoV-2 assays in the vaccinated group using receptor-binding domain antibody assays (RBD Ab assay), neutralizing antibody assay (nAb assay), and interferon-gamma release assay (IGR assay). We also compared the performance of the SARS-CoV-2 assays based on vaccine type in a large population. We collected 1851 samples from vaccinated individuals with vector, mix-and-match (MM), and mRNA vaccines. The performance of the RBD Ab assays was assessed by SARS-CoV-2 IgG II Quant (Abbott Laboratories, Sligo, Ireland), SARS-CoV-2 IgG (Beckman Coulter, CA, USA), and anti-SARS-CoV-2 S (Roche Diagnostics GmbH, Mannheim, Germany). The nAb assay was assessed by cPass SARS-CoV-2 neutralization antibody detection kits (GenScript, NJ, USA). The IGR assay was assessed by QuantiFERON (Qiagen, Venlo, The Netherlands). Median values of the RBD Ab assays and nAb assay sequentially increased after the first and second vaccinations. RBD Ab assays and nAb assay showed very strong correlations. The median values of the RBD Ab, nAb, and IGR were higher in the mRNA vaccine group than in the vector and MM vaccine groups. The agreement and correlation among the RBD Ab assays, nAb assay, and IGR assay were higher in the mRNA vaccine group than in the vector and MM vaccine groups. We compared the performance of the RBD Ab assay, nAb assay, and IGR assay based on the vaccine types using the RBD Ab, nAb, and IGR assays. This study provides a better understanding of the assessment of humoral and cellular immune responses after vaccination.

A novel three-biomarker signature (CA-62, CEA, CYFRA 21-1) and early-stage NSC lung cancer detection.

3038 Background: A double-blind clinical study was conducted on 304 clinically verified blood serum samples, including 141 non-small cell lung cancer (NSCLC), 133 healthy volunteers and 30 patients with chronic obstructive pulmonary disease (COPD). The objectives of the study were: The assessment of various cancer markers and their possible combinations for detection of early (I-II) stages of non-small cell lung cancer (NSCLC) and evaluation of the best cancer markers panel as a pre-screening tool for NSCLC before Low-Dose CT scan. Methods: Quantitative measurement of various tumor markers for serum samples was carried out using electrochemiluminescent immunoassays Elecsys CA-125, ELECSYS CA 19-9, ELECSYS CYFRA 21-1 and ELECSYS SCC (COBAS, Roche Diagnostics GmbH, Germany, EU), enzyme immunoassay CA15-3-ELISA-BEST, CEA-ELISA-BEST, NSE-ELISA-BEST (JSC "VECTOR-BEST", RF) and immunochemiluminescent analysis CLIA-CA-62 (LLC "JVS Diagnostics", RF). The results were compared to the literature data for other biomarkers and their panels. Results: Unlike other tumor markers, which are expressed proportionally to tumor size, the marker for epithelial carcinomas CA-62 is expressed from Stage I and demonstrated the highest diagnostic results in detecting early stages (I-II) of NSCLC. The biomarkers CYFRA 21-1 and SCC do not have sufficient sensitivity and specificity to diagnose asymptomatic lung cancer. However, the addition of cytokeratin 19 CYFRA 21-1 to glycoproteins CA-62 and CEA increases the specificity by eliminating false positive results, which significantly improves the diagnostic value of the cancer marker signature (CA-62, CEA and CYFRA 21-1): 100% Specificity, 93% Sensitivity, 100% PPV, 94% NPV and 94% test accuracy. Conclusions: The results of the study demonstrated that using the biomarkers signature (CA-62, CEA and CYFRA 21-1) allows increasing the specificity of CT scan for patients with suspicious abnormalities on the tomogram, improving the interpretation of visualized localized lesion, and improving the accuracy of differential diagnosis at detecting early stages of LC up to 94%. In the prospective, adding cancer markers panel signature (CA-62, CEA and CYFRA 21-1) to the current lung cancer risk assessment system as a pre-screening tool for LDCT may improve the quality of early-stage Lung cancer detection by significant increasing the sensitivity and by reducing the proportion of false positive results. [Table: see text]

Diagnostic significance of CA-62 cancer antigen for early detection and differential diagnosis of non-small cell lung cancer: results of the blind clinical trials

Background. The combination of several diagnostic methods is used to predict treatment outcomes, assess overall survival, and increase the positive predictive value of detecting malignant lung and bronchial tumors.
 Aim. To evaluate the diagnostic value of the CLIA-СА-62 chemiluminescence immunoassay reagent kit for the detection of early (IaIIb) and advanced (IIIac) stages of lung cancer (LC) in a double-blind clinical study and to assess the use of the CA-62 cancer antigen as a supportive decision-making tool in LC diagnosis in patients with suspicious changes on the tomogram or as a tool for pre-screening of LC prior to computed tomography (CT) to increase diagnostic sensitivity in the detection of early (I and II) stages of LC.
 Materials and methods. A blinded clinical study was conducted on 304 clinically verified serum samples, including 141 samples from patients with non-small cell LC (NSCLC), 133 healthy volunteers, and 30 chronic obstructive pulmonary disease patients. Quantification of other well-known tumor markers used in the diagnosis of LC (CEA, CA-125, CA 15-3, CA 19-9, CYFRА 21-1, NSE, and SCC), as well as the CA-62 marker in all serum samples was performed using electrochemiluminescent immunoassay Elecsys CA-125, ELECSYS CA 19-9, ELECSYS CYFRА 21-1 and ELECSYS SCC (COBAS, Roche Diagnostics GmbH, Germany, EU), enzyme-linked immunoassay CA 15-3-ELISA-BEST, CEA-ELISA-BEST, NSE-ELISA-BEST (AO Vector-Best, Russia) and chemiluminescent immunoassay CLIA-СА-62 (JVS Diagnostics, Skolkovo, Moscow, Russia).
 Results. CA-62 glycoprotein showed the highest level of expression at stage I NSCLC (12 745 U/mL) compared to other tumor markers studied and remained very high at the later stages of cancer: stage II (11 261 U/mL) and stage III (10 220 U/mL). A comparative analysis of the ROC curves of the most promising tumor markers CEA, CYFRA 21-1, SCC, and CA-62 for the entire NSCLC cohort versus all healthy volunteers and patients with chronic obstructive pulmonary disease showed a significant difference in the area under the curve between CA-62 (AUC 0.981) and other markers: CEA (AUC 0.84) CYFRA 21-1 (AUC 0.753)SCC (AUC 0.682). When detecting early stages (I and II) of NSCLC, a comparison of the sensitivity of the studied tumor markers showed the following pattern: CA-62 (92%)CEA (37%)CYFRA 21-1 (9%) and SCC (9%)NSE (4.5%)CA-125 (3%)CA 15-3 (1.5%)CA 199 (1%). In contrast to the CEA, CA 15-3, CA-125, NSE, CA 19-9, CYFRA 21-1, and SCC tumor markers, which are expressed proportionally to tumor growth, the epithelial carcinoma marker CA-62 showed the highest diagnostic indicators in the detection of LC early stages (III): sensitivity 92.5%, specificity 96.3%, positive predictive value 91.2%, NPV 97%, with 95% accuracy of LC detection with biopsy.
 Conclusion. The study results showed that in order to increase the specificity of computed tomography in diagnosing LC in patients with suspicious lesion on the CT scan on the tomogram, the use of the carcinoma-specific marker CA-62 can improve the interpretation of the localized focus visualized and increase the accuracy of differential diagnosis at the early stages of LC to 96%, thus contributing to an increase of the overall survival among patients with lung cancer. Of the entire panel of markers, only glycoprotein CA-62 showed a strong correlation with histology (kappa 0.91) in identifying the malignant process with inconclusive results of low-dose CT (LDCT). In the future, introducing the CA-62 marker to the current system for assessing the LC risk as a pre-screening for LDCT can improve the detection of early LC by reducing false-positive results. Once introduced into existing screening programs, it can help significantly reduce the number of patients who need LDCT, decreasing the workload of LDCT and reducing radiation exposure.

Potential clinical application of the cancer antigen CA‑62 for differential diagnosis of prostate cancer and benign prostatic hyperplasia at the elevated prostate‑specific antigen

Purpose of the study. Evaluation of the diagnostic characteristics of the CA‑62 marker for epithelial carcinomas for detecting early‑stage prostate cancer in a double‑blind clinical study. This study is also focused on the possibility of using the CA‑62 antigen as an auxiliary tool for decision‑making in prostate cancer diagnosis.Patients and methods. A blinded clinical study was conducted on 325 clinically verified blood serum samples. This includes 144 prostate cancer samples, 79 generally healthy volunteers‑men and 102 samples from patients with benign prostatic hyperplasia (BPH). Quantitative determination of the total and free prostate specific antigen (PSA) levels, as well as the CA‑62 marker of serum samples was performed using the electrochemiluminescent immunoassay ECLIA Elecsys Total and Free PSA (COBAS, Roche Diagnostics GmbH, Germany, EU) and the chemiluminescent immunoassay CLIA‑CA‑62 (JVS Diagnostics LLC, Moscow, RF).Results. A comparison of the CA‑62 level with the results for total and free PSA, as well as other diagnostic methods (PCA3, PHI) for the analysis of the BPH and prostate cancer groups was performed. The results show that the CA‑62 marker has the highest PPV (94.4 %) and NPV (93.1 %). This may increase the reliability of the decision related to the presence of PC and be used by doctors as an argument as an argument for a prostate biopsy referral. It has been demonstrated that using the novel cancer marker CA‑62 makes it possible to detect up to 90 % of the early‑stage prostate cancer with 97.2 % specificity (AUC = 0.969).Conclusion. Using the CA‑62 marker as an auxiliary diagnostic method within the PSA “grey zone” (from 2.5 to 10 ng/ml) made it possible to significantly increase the accuracy of detecting the PC early stages at biopsy up to 93.1 %. It will help the doctors to effectively differentiate between prostate cancer and benign prostatic hyperplasia.

Frequency of Anemia and Iron Deficiency Anemia in Patients with Helicobacter Pylori Infection: A Cross-Sectional Study

Objective: To determine the frequency of anemia and iron deficiency anemia in patients with Helicobacter pylori infection. Study design: Cross-sectional study Place and Duration: Department of Medicine, Liaquat University Hospital, Hyderabad from May to November 2019 Methodology: We included a total of 140 patients with anemia, either inpatients or outpatients at Liaquat University Hospital in Hyderabad's department of medicine. We acquired demographic information, such as name, gender, and age. The study involved everyone who meets the inclusion requirement. Anemia's brief history, its duration, the colour of the stools (melena black faeces), and the ferritin level were all noted. The cubital vein of either upper arm was used to collect 10 ml of blood, which was then sent to the lab for analysis of anaemia, ID, and H. Pylori detection. A multiparameter cell counter Sysmex k 1000 hematology analyzer was used to determine the amounts of haemoglobin (Hb) and hematocrit (Hct) in the CBC (Tao electronics, Japan). Serum iron levels were evaluated using a Hitachi 917 biochemistry automated analyzer, and ferritin concentrations were determined using a Cobas e411 hormones auto-analyzer from Roche Diagnostics GmbH in Mannheim, Germany (Roche Diagnostics GmbH, Mannheim, Germany). The researcher himself entered all the data on a proforma that had been previously created. Results: Patients' average ages ranged from 40.44± 14.268 years, and their haemoglobin levels were 8.77 ± 3.14. Patients' serum ferritin levels were 10.5 ±63.21 and their BMI was 24.5 ±3.21 respectively. Of the H. pylori patients, there were 70 (50%) anaemic patients and 70 (50%) with normal Hb levels. In this study, 102 individuals (72.9%) had normal serum profiles while 38 patients (27.1%) had iron deficiency anemia. Conclusion: According to our research, treating H. pylori infection may help IDA patients who are infected with the parasite especially those who have moderate to severe anaemia. Individuals with H. pylori infection had a significant prevalence of anaemia and IDA, and both conditions were linked to patients' ages and several behavioural traits Keywords: H. pylori infection, anemia, iron deficiency anemia

Changes of Plasma Blood Ammonia Levels of Chinese Healthy People and the Establishment of Reference Intervals

Blood ammonia detection is used for the diagnosis or differential diagnosis of various hepatitis virus infections, severe liver cirrhosis, and hepatic encephalopathy. It is also one of the important indexes reflecting liver coma, Reyes syndrome, and other diseases. However, blood ammonia changes rapidly with time. If samples are not sent and detected in time, the results will be wrong, resulting in clinical misdiagnosis and life danger to patients. The purpose of this paper is to explore the change of blood ammonia with time and establish its reference interval. For this study, 228 healthy patients (111 males and 117 females) were selected who underwent physical examination at the Health Management Center of the Second Xiangya Hospital of Central South University from April to May 2021. The blood ammonia detection kit (colorimetric method) produced by Roche Diagnostics GmbH of Germany was used for detection on the Roche cobas c702 automatic biochemical analyzer. After eliminating outliers from the obtained test results, they were grouped according to gender and age, and SPSS 26.0 software was employed to statistically analyze the blood ammonia test results. The differences in blood ammonia levels at each detection time were statistically significant (p < 0.05). The differences in blood ammonia levels between male and female subjects at 1 hour, 2 hours, and 3 hours were statistically significant (p < 0.05), but all ages saw no statistically significant difference in blood ammonia levels between segments (p > 0.05). The blood ammonia levels of each detection time and different genders showed a normal distribution. Therefore, it is necessary to take the 95% (X ± 1.96S) results of both sides as the reference interval according to the detection time and gender, and establish the reference intervals. The 1-hour blood ammonia reference interval for healthy men in Changsha is 15.8 - 47.5 μmol/L, for healthy women it is 12.4 - 39.6 μmol/L; the 2-hour blood ammonia reference interval for healthy men is 22.3 - 56.5 μmol/L, and for healthy women it is 19.1 - 48.0 μmol/L; the reference interval of 3-hours blood ammonia for healthy men is 27.9 - 65.7 μmol/L, and for healthy women it is 24.6 - 56.7 μmol/L. There are differences in blood ammonia levels between men and women at different detection times in Changsha. A reference interval suitable for blood ammonia in healthy individuals in the region should be established according to the detection time and gender, so as to provide better relevant evidence for clinical diagnosis.

Plasma Levels of Secretory Calreticulin among HIV Infected Patients in Bauchi State

Objectives: Human immunodeficiency virus (HIV) is a lentivirus that causes HIV infection and over time acquired immunodeficiency syndrome. The objective was to evaluate plasma levels of Calreticulin (CALR) in HIV positive and negative individuals. Methods: A total of 164 participants were recruited and divided into four groups according to their viral load after estimation and individuals recruited as controls. Viral load was estimated in 1.1 ml of plasma according to manufacturers (COBAS Taqman HIV-1 Quantitative Test, v2.0, Roche Diagnostic GmbH, Germany). Plasma levels of CALR were analysed using ELISA (ELISA-Biotuva Life Sciences, UK). Results: Result revealed a significant difference (p = 0.00) between the CALR levels of individuals with low viral load and individuals with unsuppressed viral load, suppressed viral load, and controls. A significantly (p = 0.017) higher plasma CALR in individuals with unsuppressed viral load compared to those with suppressed viral load was found. No differences between plasma CALR levels of individuals with unsuppressed viral load and that of controls. Conclusion: Our study showed up regulation of plasma CALR in HIV-infected patients on long term antiretroviral therapy. We believe and from existing literature this up regulation is due partly if not solely to the oxidative stress and endoplasmic reticulum stress caused by the antiretroviral agents.

Multicenter performance evaluation of the Elecsys HCV Duo immunoassay.

The diagnostic accuracy of the Elecsys® HCV Duo antigen-antibody combination immunoassay (Roche Diagnostics GmbH) was evaluated for the detection of hepatitis C virus (HCV) infection, versus commercially available comparators. This multicenter study (August 2020-March 2021) assessed the specificity of the Elecsys HCV Duo immunoassay and comparator assays in blood donor and routine clinical laboratory samples; sensitivity was determined in confirmed HCV-positive samples and seroconversion panels. The Elecsys HCV Duo immunoassay was compared with the Monolisa HCV Ag-Ab ULTRA V2, Murex HCV Ag/Ab Combination and ARCHITECT HCV Ag assays, as well as nucleic acid testing (NAT). The antibody (anti-HCV) module of the Elecsys HCV Duo immunoassay was compared with the Elecsys Anti-HCV II, Alinity s Anti-HCV, ARCHITECT Anti-HCV and RIBA HCV 3.0 SIA assays. The specificity of the Elecsys HCV Duo immunoassay was 99.94% (95% confidence interval [CI], 99.89-99.97) and 99.92% (95% CI, 99.71-99.99) in blood donor (n=20,634) and routine clinical laboratory samples (n=2531), respectively. The specificity of the Elecsys HCV Duo immunoassay was similar or better than comparator assays. The sensitivity of the Elecsys HCV Duo immunoassay in confirmed HCV-positive samples (n=257) was 99.6%. In seroconversion panels, the Elecsys HCV Duo immunoassay detected infections earlier (2.2-21.9 days) than all but one of the comparator assays and detected HCV 1.8 days later than NAT. The Elecsys HCV Duo immunoassay shows high diagnostic accuracy, reduces the diagnostic window, and could be used when NAT is not possible.

Early carbohydrate antigen 125 as a mortality predictor in hospitalized patients with coronavirus disease 2019.

Carbohydrate antigen 125 (CA125) is an indicator of inflammation, immune response, and impaired cardiac function. The aim was to investigate whether CA125 behaves as a biomarker of severity and poor clinical outcomes in hospitalized patients with coronavirus disease 2019 (COVID-19). Serum CA125 [Elecsys CA125 II assay-(Roche Diagnostics GmbH)] was measured in stored biobank samples from COVID-19 hospitalized patients between 01 March 2020 and 17 October 2021. Multiple logistic regression models were built to explore the association between CA125 and clinical outcomes [in-hospital all-cause mortality, need for invasive mechanical ventilation (IMV), or non-invasive respiratory support (non-IRS)], estimating odds ratios (ORs; 95% CI). The gradient of risk of CA125 was evaluated by fractional polynomials. A total of 691 patients were included, median age of 63 years (50-76), men (57.2%), with high comorbidity. At admission, 85.8% had pneumonia. Median CA125 was 10.33 U/ml (7.48-15.50). The in-hospital mortality rate was 7.2%. After adjusting for confounding factors, CA125 ≥ 15.5 U/ml (75th percentile) showed an increased risk of death [OR 2.85(1.21-6.71)], as age ≥ 65 years, diabetes, and immunosuppression. Furthermore, CA125 as a continuous variable was positive and significantly associated with the risk of death after multivariate adjustment. The mean hospital stay of the patients with CA125 ≥ 15.5 U/ml was longer than the rest of the study population. CA125 in the first 72 h of hospital admission seems a useful biomarker of mortality in hospitalized patients with moderate-severe COVID-19. If our findings are confirmed, the wide availability of this biomarker would make easy its widespread implementation in clinical practice.

Effectiveness of the modified scheme for treatment of Н.pylori-associated peptic ulcer of the duodenum in children

Background. None of the known regimens for the treatment of H.pylori-associated peptic ulcer of the duodenum (PUD) is 100 % effective. The purpose was to evaluate the effectiveness of the modified comprehensive therapy for Н.pylori-associated peptic ulcer of the duodenum in children. Materials and methods. The study included 66 patients aged 7 to 18 years with Н.pylori-associated PUD in the acute stage. Serum vitamin D levels were measured using the electrochemiluminescence method (Roche Diagnostics GmbH, Mannheim, Germany). The classical method of stool culture was used. The children were divided into three groups: the first — 20 patients who received protocol eradication therapy, the second — 23 children who received modified comprehensive therapy with the inclusion of vitamin D3, the third — 23 patients who received modified comprehensive therapy with the inclusion of vitamin D3 and a synbiotic. Results. The average length of stay in the hospital for patients who received modified therapy with the inclusion of vitamin D3 was reduced by 18.8 %, and in those who received synbiotic additionally — by 23.8 % compared to the first group. During the endoscopic examination conducted after 1 month, the presence of an elongated red scar at the site of the ulcer defect was observed significantly more often in group I (65.5 %) than in groups II (47.8 %) and III (39.1 %), χ2 = 3.87, p &lt; 0.05; after 1 year, 33.3 % of children in group I, 17.3 % in group II, and 8.7 % in group III (p &lt; 0.01) had a recurrence of the disease. The determination of the main indicators for the effectiveness of PUD treatment using a modified scheme showed that the relative risk of disease recurrence will decrease by 0.42 times (χ2 = 3.96, p &lt; 0.05) when vitamin D3 preparation is included in the treatment scheme and by 0.52 times (χ2 = 4.06, p &lt; 0.01) when using vitamin D3 and a synbiotic. Conclusions. 1. Children with Н.pylori-associated peptic ulcer of the duodenum have vitamin D3 deficiency and various disturbances of the intestinal microbiota, which can complicate the course, increase the chronicity of the process, and reduce the effectiveness of anti-Helicobacter therapy. 2. The use of a modified comprehensive treatment for duodenal ulcer associated with H.pylori leads to a reduction in the duration of the main clinical manifestations of the disease, the duration of inpatient treatment of sick children and a decrease in the frequency of recurrence of the disease.

Analytical and clinical performance of a rapid magnetic immunochromatographic assay for N-terminal pro-B-type natriuretic peptide detection

ObjectiveGoldMag® immunochromatography is a novel point-of-care immunoassay platform utilizing quantitative determination of magnetic signals. A N-terminal pro-B-type natriuretic peptide (NT-proBNP) assay has been developed on this system. The objective of this study was to determine whether GoldMag® immunochromatography for NT-proBNP test fulfils the basic requirements of analytical and clinical performance so as to be used as a point-of-care test. MethodsAnalytical performance was performed for the linearity, sensitivity, precision, interference and recovery by Clinical and Laboratory Standards Institute guidelines (CLSI). Clinical evaluation was performed by testing 206 residual serum samples and comparing with Elecsys® proBNP assay (Roche Diagnostics GmbH, Mannheim, Germany). ResultsGoldMag® immunochromatography was able to differentiate concentrations of NT-proBNP from 0.000 to 8,000 pg/mL with a correlation coefficient R2 = 0.9914 and a limit of detection (LOD) of 43 pg/mL. According to statistics, this system exhibited high precision, high recovery, and no interference with lipemia, bilirubinemia, and hemolysis. Correlation analysis showed that the two assays yielded a good correlation with R2 of 0.9860. Bland-Altmananalysis showed a bias offset of 28.98 pg/mL with 95% confidence interval from 6.056 to 72.74 pg/mL for NT-proBNP. ConclusionGoldMag® immunochromatography system for NT-proBNP detection provides high accuracy, reliability, and easy operation. It can be used as a point-of-care test with a great application value in clinical management of chronic and acute heart failure patients.

Diagnostic performance of automated plasma amyloid-β assays combined with pre-analytical immunoprecipitation

BackgroundMeasurements of the amyloid-β (Aβ) 42/40 ratio in blood plasma may support the early diagnosis of Alzheimer’s disease and aid in the selection of suitable participants in clinical trials. Here, we compared the diagnostic performance of fully automated prototype plasma Aβ42/40 assays with and without pre-analytical sample workup by immunoprecipitation.MethodsA pre-selected clinical sample comprising 42 subjects with normal and 38 subjects with low cerebrospinal fluid (CSF) Aβ42/40 ratios was studied. The plasma Aβ42/40 ratios were determined with fully automated prototype Elecsys® immunoassays (Roche Diagnostics GmbH, Penzberg, Germany) by direct measurements in EDTA plasma or after pre-analytical Aβ immunoprecipitation. The diagnostic performance for the detection of abnormal CSF Aβ42/40 was analyzed by receiver operating characteristic (ROC) analysis. In an additional post hoc analysis, a biomarker-supported clinical diagnosis was used as a second endpoint.ResultsPre-analytical immunoprecipitation resulted in a significant increase in the area under the ROC curve (AUC) from 0.73 to 0.88 (p = 0.01547) for identifying subjects with abnormal CSF Aβ42/40. A similar improvement in the diagnostic performance by pre-analytical immunoprecipitation was also observed when a biomarker-supported clinical diagnosis was used as a second endpoint (AUC increase from 0.77 to 0.92, p = 0.01576).ConclusionsOur preliminary observations indicate that pre-analytical Aβ immunoprecipitation can improve the diagnostic performance of plasma Aβ assays for detecting brain amyloid pathology. The findings may aid in the further development of blood-based immunoassays for Alzheimer’s disease ultimately suitable for screening and routine use.

A randomized, double-blind, single-dose, parallel phase I clinical trial to compare the bioequivalence, immunogenicity, and safety of bevacizumab biosimilar and bevacizumab in healthy Chinese subjects

ABSTRACT Background Bevacizumab, a humanized monoclonal antibody against VEGF, can be used as a target therapy for colorectal cancer. A phase I clinical trial was conducted to compare the bioequivalence, immunogenicity, and safety of bevacizumab biosimilar (Chia Tai Tianqing Pharmaceutical Group Co., Ltd.) and Bevacizumab (Roche Diagnostics GmbH) in healthy Chinese males. Research design & method Healthy Chinese subjects (N = 98) were randomly divided into two groups. A single-dose bevacizumab biosimilar or Bevacizumab was given per cycle. Plasma drug concentrations were detected by liquid chromatography–tandem mass spectrometry (LC-MC/MS) assay. We detected the levels of anti-drug antibody (ADA) to evaluate drug immunogenicity and the safety of drugs throughout the study. Results The geometric mean ratios (GMRs) of AUC0-t, Cmax, and AUC0-∞ for bevacizumab biosimilar and Bevacizumab were 96.27%, 93.69%, and 97.01%, respectively. The 90% CIs were all within 80–125%, meeting the bioequivalence standards. The levels of ADA were similar. In addition, the two drugs both demonstrated excellent safety in the trial. Conclusion This study showed that bevacizumab biosimilar and Bevacizumab had similar pharmacokinetics (PK) parameters and safety in healthy Chinese subjects.

Prognostic Utility of Procalcitonin, Presepsin, and the VACO Index for Predicting 30-day Mortality in Hospitalized COVID-19 Patients.

BackgroundBiomarkers and clinical indices have been investigated for predicting mortality in patients with coronavirus disease (COVID-19). We explored the prognostic utility of procalcitonin (PCT), presepsin, and the Veterans Health Administration COVID-19 (VACO) index for predicting 30-day-mortality in COVID-19 patients.MethodsIn total, 54 hospitalized COVID-19 patients were enrolled. PCT and presepsin levels were measured using the Elecsys BRAHMS PCT assay (Roche Diagnostics GmbH, Mannheim, Germany) and HISCL Presepsin assay (Sysmex, Kobe, Japan), respectively. The VACO index was calculated based on age, sex, and comorbidities. PCT and presepsin levels and the VACO index were compared using ROC curve, Kaplan–Meier method, and reclassification analysis for the 30-day mortality.ResultsROC curve analysis was used to measure PCT and presepsin levels and the VACO index to predict 30-day mortality; the optimal cut-off values were 0.138 ng/mL for PCT, 717 pg/mL for presepsin, and 12.1% for the VACO index. On Kaplan–Meier survival analysis, hazard ratios (95% confidence interval) were 15.9 (4.1-61.3) for PCT, 26.3 (6.4-108.0) for presepsin, and 6.0 (1.7-21.1) for the VACO index. On reclassification analysis, PCT and presepsin in addition to the VACO index significantly improved the prognostic value of the index.ConclusionsThis study demonstrated the prognostic utility of measuring PCT and presepsin levels and the VACO index in COVID-19 patients. The biomarkers in addition to the clinical index were more useful than the index alone for predicting clinical outcomes in COVID-19 patients.

Vitamin D levels in children with celiac disease

Background. A significant increase in understanding of the role of vitamin D in the body, more effective detection of celiac disease, and the need to monitor the health of children against the background of long-term adherence to a gluten-free diet were prerequisites for our study. The study was aimed to analyze the level of vitamin D in children with celiac disease. Materials and methods. The results of the examination of 29 children aged from 6 months to 18 years with a verified diagnosis of celiac disease were analyzed. Serum vitamin D levels were measured by the electrochemiluminescent method (Roche Diagnostics GmBH, Mannheim, Germany). The results of vitamin D supplementation in patients with celiac disease were compared with the control group of 30 healthy children aged from 1 to 18. Mathematical processing of the material included a standard algorithm for statistical research using Microsoft Excel 2016, Attestat. Results. Among the patients included in the study, typical celiac disease was found in 24 (82.7 %) cases, which is 4.8 times more common than atypical — in 5 (17.4 %) children. The gastrointestinal symptoms dominated in a clinical picture. Manifestation of the disease in most patients was observed in the first year of life — in 17 (58.6 %) cases, in 7 (24.1 %) patients aged from 1 to 3 years, and only in 5 (17.4 %) children older than 3 years. The average rate of vitamin D in children with celiac disease was probably lower than in healthy children and accounted for 24.4 ± 1.2; 21.2 [16.45–35.21] ng/ml. The number of children with normal vitamin D content is the highest among young patients, while the frequency of vitamin D deficiency is the lowest. The median serum vitamin D in patients on a gluten-free diet was 1.4 times higher (p &lt; 0.05) than in the acute period, but 1.3 times lower (p &lt; 0.05) than in the control group. Adherence to a gluten-free diet leads to increased levels of vitamin D but does not allow reaching the level in healthy children. Conclusions. Vitamin D deficiency is registered in children with celiac disease. All patients with celiac disease, regardless of the stage of the disease and adherence to a gluten-free diet, need to be monitored for vitamin D levels.

Sustained but Declining Humoral Immunity Against SARS-CoV-2 at 9 Months Postvaccination With BNT162b2: A Prospective Evaluation in 309 Healthy Individuals.

The sustainability of coronavirus 19 (COVID-19) vaccine-induced immunity against severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is critical to be determined to inform public health decisions on vaccination programs and prevention measures against COVID-19. The aim of the present study was to prospectively evaluate the kinetics of neutralizing antibodies (NAbs) and anti-S-receptor binding domain (RBD IgGs) against SARS-CoV-2 after full vaccination with the BNT162b2 mRNA vaccine for up to 9 months in healthy individuals (NCT04743388). The assessments were performed at the following time points after the second vaccination: 2 weeks, 1 month, 3 months, 6 months, and 9 months. The measurements were performed with the GenScript’s cPassTM SARS-CoV-2 NAbs Detection Kit (GenScript, Inc.; Piscataway, NJ) and the Elecsys Anti-SARS-CoV-2 S assay (Roche Diagnostics GmbH; Mannheim, Germany). Three hundred nine participants with a median age of 48 years were included. A gradual decline in both NAbs and anti-S-RBD IgGs became evident from 2 weeks to 9 months postvaccination. Both NAbs and anti-S-RBD IgGs levels were significantly lower at 9 months compared with the previous timepoints. Interestingly, age was found to exert a statistically significant effect on NAbs elimination only during the first-trimester postvaccination, as older age was associated with a more rapid clearance of NAbs. Furthermore, simulation studies predicted that the median NAb value would fall from 66% at 9 months to 59% and 45% at 12 and 18 months postvaccination, respectively. This finding may reflect a declining degree of immune protection against COVID-19 and advocates for the administration of booster vaccine shots especially in areas with emerging outbreaks.