High-speed centrifugation rather than Lipoclear reagent can be used for removing the interference of lipemia on serological tests of infectious diseases: AIDS, hepatitis B, hepatitis C, and syphilis by chemiluminescent microparticle immunoassay.

The aim of this study was to investigate the interference of lipemia on measurement of HBsAg, anti-HBs, HBeAg, anti-HBe, anti-HBc, anti-HCV, HIV Ag/Ab, and anti-TP in serum by chemiluminescent microparticle immunoassay (CMIA) and compare lipemia removing performance between high-speed centrifugation and Lipoclear reagent. Mixed native serum samples (NSs) and hyperlipemia serum samples (HLS) were prepared for the investigated parameters. The levels of these parameters in NS and HLS were determined by CMIA on an Abbott ARCHITECT i2000SR immunoassay analyzer. HBsAg, anti-HBs, and anti-TP were affected with relative bias >12.5% (acceptable limit) when the level of triacylglycerol (TG) was higher than 27.12 mmol/L in HLS. Clinically unacceptable bias were observed for HBeAg and anti-HBe in HLS with TG higher than 40.52 mmol/L. However, anti-HCV and HIV Ag/Ab were not interfered in severe lipemia with TG < 52.03 mmol/L. In addition, the Lipoclear reagent did not reduce the interference of lipemia with relative bias from -62.50% to -18.02%. The high-speed centrifugation under the optimized condition of 12 000g for 10 min successfully removed the interference of lipemia with relative bias from -5.93% to 0% for HBsAg, anti-HBs, HBeAg, anti-HBe, anti-HBc, and anti-TP. To conclude, high-speed centrifugation can be used for removing the interference of lipemia to measure HBsAg, anti-HBs, HBeAg, anti-HBe, anti-HBc, and anti-TP. Accordingly, a standardized sample preanalytical preparation of the patients and other screening participants as well as a specimen examination procedure for removing lipemia interference on the serological tests was recommended.

Antibody responses to COVID-19 vaccines in older adults.

Because of the senescence of the immune system, antibody response to the COVID-19 vaccines may differ from older to younger adults. The study aim compares the titers of SARS-CoV-2 IgG antibody of patients ≥60 years who received three doses of CoronaVac vaccine and those who received two doses of CoronaVac+1 dose of Pfizer-BioNTech after 1 month of the last vaccination. Patients ≥60 years who received the CoronaVac vaccine between March 1, 2021, and April 30, 2021, who did not have COVID-19 disease before the first dose of vaccination and were negative for COVID-19 antibodies, whose antibodies were tested before the third dose of vaccination, and who did not have any COVID-19 disease during the follow-up were included. The demographic characteristics and comorbidities of patients were recorded. An immunofluorescence assay (IFA) fast test and a chemiluminescent microparticle immunoassay (Abbott) were used to measure SARS-CoV-2 quantitative antibody levels at the first month after the third-dose vaccine. Totally 81 patients, 41 patients in third dose of the CoronaVac group (female:male 18:23, mean age 69.4 ± 8.5), and 40 patients in third dose of the Pfizer-BioNTech group (female:male 15:25, mean age 69.9 ± 9.1) were included. The patients' comorbidities in the groups were similar. The titers of IgG antibodies to SARS-CoV-2 measured according to both IFA and Abbott Kit at first month the third dose vaccination was significantly higher in the Pfizer-BioNTech group (p ≥ 0.001, p = 0.012, respectively). The results report that the formed immunity in the first month after the two doses of CoronaVac+1 dose Pfizer-BioNTech vaccine was higher than three doses of CoronaVac vaccine in older adults.

Quantitation of antibodies against SARS-CoV-2 spike protein after two doses of CoronaVac in healthcare workers.

Quantitation of antibodies to the spike protein of severe acute respiratory syndrome coronavirus 2  (SARS-CoV-2) was performed for the detection of adaptive immune response in healthcare workers (HCWs) vaccinated with CorovaVac. We prospectively recruited HCWs from a university hospital in Turkey. Serum samples from 1072 HCWs were obtained following 28 days of the first, and 21 days of the second dose. Detection and quantitation of SARS-CoV-2 antispike antibodies were performed by the chemiluminescent microparticle immunoassay (SARS-CoV-2 IgG II Quant; Abbott). Results greater than or equal to the cutoff value 50.0 AU/ml were reported as positive. After the first dose, antispike antibodies were detected in 834 of 1072 (77.8%) HCWs. Seropositivity was higher among females (84.6%) than males (70.6%) (p < 0.001) and was found to be highest in both women and men between the ages of 18-34. After the second dose, antibodies were detected in 1008 of 1012 (99.6%) HCWs. Antibody titers were significantly higher in those who had coronavirus disease-2019 before vaccination than those who did not (p < 0.001). Antibody positivity and median antibody titers were significantly less in HCWs with chronic diseases compared to those without (p < 0.05 and p < 0.001, respectively). In conclusion, our findings indicated that a relatively high frequency (99.6%) of humoral immunity was produced in HCWs aged 18-59 after two doses of CoronaVac. Quantitation of antibodies may help facilitate longitudinal monitoring of the antibody response, which will be especially useful in deciding the dose of the vaccine in vulnerable groups such as those over 60 years of age and those with chronic diseases.

Comparison and evaluation of Abbott chemiluminescent microparticle immunoassay and ChIVD light-initiated chemiluminescent assay in the detection of Treponema pallidum antibody.

BACKGROUND: Laboratory tests play an important role in the diagnosis of syphilis. This study aimed to compare and assess the performance of the Abbott chemiluminescent microparticle immunoassay (CMIA) and the ChIVD light-initiated chemiluminescent assay (LICA) in the detection of Treponema pallidum (TP) antibody. METHODS: A total of 10 498 serum samples were detected with two assays, and the Treponema pallidum particle agglutination assay (TPPA) and recombinant immunoblot assay (RIBA) methods were used for confirmation. The sensitivity, specificity, positive predictive value, and negative predictive value of the Abbott CMIA and ChIVD LICA were calculated. The coincidence rate between two assays was also evaluated. The causes of false positive and false negative of two assays were studied. RESULTS: For the Abbott CMIA and ChIVD LICA, the sensitivity was 94.44% and 98.15%, the specificity was 99.89% and 99.81%, the positive predictive value was 93.29% and 88.83%, and the negative predictive value was 99.91% and 99.97%, respectively. The coincidence rate between Abbott CMIA and ChIVD LICA was 99.26%, and κ value was .790. The disease of infertility, hypertensive disease, liver disease, and cancer were the common causes of false positive in both assays, while infertility was also the main reason lead to false negative. CONCLUSION: Our results demonstrated that the Abbott CMIA and ChIVD LICA generally had high sensitivity and specificity and therefore may be suitable for the detection of TP antibody and screening for syphilis.

Evaluation of three automated Treponema pallidum antibody assays for syphilis screening.

The accuracy of the test is critical for the syphilis serology diagnosis. This study aims to evaluate the values of the Elecsys syphilis assay, the Architect syphilis assay, and the Mindray syphilis assay, as syphilis screening tests for pregnant women and patients with syphilis or other diseases. A reverse algorithm was used for the syphilis serology diagnosis. Serum samples (n = 584) were tested with three automated screening assays. All reactive sera by one, two, or three screening assays were further analyzed with the tolulized red unheated serum test (TRUST). Inconsistent results were confirmed by the Treponema pallidum particle agglutination assay (TPPA). The final patient diagnosis was made according to the results of syphilis serology, clinical evidence, and past medical history. The sensitivity, specificity, accuracy, and kappa value of each assay were as follows: for the Elecsys syphilis assay, 100.0%, 98.5%, 98.6%, and 0.927, respectively; for the Architect syphilis assay: 100.0%, 94.5%, 95.0%, and 0.770; and for the Mindray syphilis assay: 100.0%, 97.0%, 97.3%, and 0.862. The McNemar test showed that there were significant differences in the performance between the Elecsys syphilis assay and the Architect syphilis assay (P < 0.001), and between the Mindray syphilis assay and the Architect syphilis assay (P = 0.001). Our study demonstrated that three automated Treponema pallidum antibody assays generally showed high sensitivities and specificities, and so, they are suitable for use in screening for syphilis. The performances of the Elecsys syphilis assay and the Mindray syphilis assay are superior to Architect syphilis assay.

Diagnostic reliability of Architect anti-HCV assay: Experience of a tertiary care hospital in India.

BACKGROUND & AIMS: Anti-HCV assays are prone to false positive results. Thus, accurate detection of HCV infection is critical for the timely therapeutic management. This study ascertained the reliability of Architect anti-HCV assay (Abbott) and to estimate the agreement of this assay with Ortho HCV 3.0 ELISA Test System with Enhanced SAVe (Ortho), HCV Tri-dot (Tri-dot) and HCV-PCR in a tertiary care setting. METHODS: A total of 78 788 consecutive sera were routinely screened for anti-HCV antibodies using Architect. All repeatedly reactive anti-HCV sera (n=1000) and anti-HCV negative sera (n=300) were tested in Ortho and in Tri-dot assays. Representative proportions of sera (n=500) with various signal-to-cut-off (S/Co) ratio were also compared with HCV-PCR. RESULTS: When Architect was compared with Ortho, Tri-dot, and HCV-PCR, the level of agreement as assessed by kappa were .26, .16, and .27 respectively. Using Latent class analysis (LCA), we found that sensitivity and specificity were 100% and 36.1% for Architect, 93.8% and 100% for Ortho and 63.8% and 100% for Tri-dot respectively. The median S/CO ratio of Architect and Ortho anti-HCV assays were significantly different between HCV-PCR positive and negative results (P<.0001). Furthermore, Architect S/CO ratio of >8 showed higher accuracy indices in both anti-HCV assays. CONCLUSIONS: Architect can be used as a screening assay because of its high sensitivity, high throughput, and short turnaround time. However, S/Co ratios of ≥1 to <8 in Architect necessitates HCV PCR to identify current infection and or EIA to distinguish true positivity from false biological positivity.

Comparison of LC-MS/MS vs chemiluminescent microparticle immunoassay in measuring the valproic acid concentration in plasma of epilepsy patients in a new perspective.

OBJECTIVES: This study was designed to compare the performance of LC-MS/MS with chemiluminescent microparticle immunoassay (CMIA) for determination of VPA in epilepsy patients in the perspective of metabolites' hepatotoxicity. METHOD: Samples were collected and then analyzed using both LC-MS/MS and CMIA. A LO2 cells (normal human hepatic cells) experiment was carried out to confirm VPA metabolites' hepatotoxicity using AST(Aspertate Aminotransferase, AST), ALT(Alanine aminotransferase, ALT) and LDH(lactate dehydrogenase, LDH) in supernate as index. RESULTS: The regression equation analysis showed as LC-MS/MS=1.0094CMIA-1.8937, with the concordance correlation coefficient of 0.9700, and the CUSUM test proved no significant deviation from linearity (P>.05). CMIA compared to LC-MS/MS gave a positive bias of 1.2 µg/mL. In LO2 experiment, VPA and its metabolites groups showed an obvious increment of AST, ALT, and LDH in supernate. CONCLUSION: The LC-MS/MS is largely consistent with the CMIA in analytical time and quantification ability for VPA, but the LC-MS/MS can simultaneously determinate VPA and its metabolites in plasma, and is also a higher cost-efficiency method in consideration of toxic metabolites monitoring. The overestimation of VPA by CMIA showed no clinical significance. The metabolites 3-OH-VPA and 5-OH-VPA damage the LO2 cells and the results presented a statistical significance (P<.05). It is vital to monitor the metabolites' concentrations for VAP's clinical safety application, and now is the occasion that laboratory and clinic consider the LC-MS/MS method as a more advantageous alternative to CMIA method in therapeutic monitoring of VPA.

Assessment of Vcheck® analyzer for rapid progesterone concentration measurement including recommendations for achieving the optimal breeding time in bitches.

Background and Aim: Serum progesterone concentration plays critical role in determining the optimal breeding time in bitches and diagnosing reproductive-related issues. This study aimed to conduct a comparative analysis of serum progesterone results obtained from commercial point-of-care immunological analyzers, namely, Vcheck®, with those obtained using chemiluminescent microparticle immunoassay (CMIA). Our overarching goal was to evaluate these analyzers' accuracy and establish standardized guidelines for optimal breeding timing. Materials and Methods: Ninety-four serum samples from bitches were analyzed using the Vcheck® analyzer and compared with CMIA. Thorough documentation included the mean, standard deviation, 95% confidence interval (CI), and minimum and maximum values of serum progesterone concentrations. Furthermore, Pearson's correlation coefficient, Lin's concordance correlation coefficient, and the bias correction factor were meticulously recorded. Results: The mean progesterone concentration measured using the Vcheck® analyzer was significantly lower than that measured using CMIA, with a mean difference of 1.26 ng/mL of serum. The Bias correction factor was 0.935, which was nearly 1.00, indicating that the line of best-fit was on the perfect line of agreement, providing insight into the measurement accuracy. Pearson's correlation coefficient, a measure of precision, was also close to 1 (0.939), confirming the reliability of the data. Furthermore, Lin's concordance correlation coefficient was 0.877, indicating a fair overall agreement between the Vcheck® and CMIA methods. These results support the validity of the Vcheck® analyzer's results. The present study was developed by aligning with established CMIA guidelines and adapting them using the range and 95% CI derived from each set of results, ensuring a standardized and rigorous approach. Conclusion: The Vcheck® analyzer provides a rapid assessment of serum progesterone concentration in bitches, with results comparable to those measured using the CMIA technique. However, when considering the use of the Vcheck® analyzer, it is recommended that the results should be interpreted carefully and the interpretation guidelines should be followed. In conclusion, Vcheck® provides a reliable and convenient method for veterinarian practitioners to measure canine progesterone levels in a clinical/hospital setting.

The Anti-SARS-CoV-2 S-Protein IgG, Which Is Detected Using the Chemiluminescence Microparticle Immunoassay (CMIA) in Individuals Having Either a History of COVID-19 Vaccination and/or SARS-CoV-2 Infection, Showed a High-Titer Neutralizing Effect.

Neutralizing antibodies plays a primary role in protective immunity by preventing severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) from entering the cells. Therefore, characterization of antiviral immunity is important for protection against SARS-CoV-2. In this study, the neutralizing effect of the anti-SARS-CoV-2 S1 protein IgG, which was detected using the chemiluminescence microparticle immunoassay (CMIA)-based SARS-CoV-2 IgG II Quant (Abbott, Waukegan, IL, USA) test in SARS-CoV-2 infected and/or vaccinated individuals, was investigated with a surrogate virus neutralization test (sVNT). In total, 120 Seropositive individuals were included in this study. They were divided into two groups: Vaccinated (n = 60) and Vaccinated + Previously Infected (n = 60). A commercial sVNT, the ACE2-RBD Neutralization Test (Dia.Pro, Milan, Italy), was used to assess the neutralizing effect. The assay is performed in two steps: screening and titration. The screening showed positive results in all seropositive samples. Low titration in 1.7%, medium titration in 5%, and high titration in 93.3% of the Vaccinated group, and medium titration in 1.7% and high titration in 98.3% of the other group, as obtained from the ACE2-RBD titration test. A strong positive and significant correlation was found between the SARS-CoV-2 IgG II Quant test and the ACE2-RBD titration test at the 1/32 titration level for both groups (p < 0.001 for both). This study shows that the SARS-CoV-2 IgG detected using the CMIA method after SARS-CoV-2 infection and/or vaccination has a high neutralizing titration by using the sVNT. In line with these data, knowledge that seropositivity determined by CMIA also indicates a strong neutralizing effect contributes to countrywide planning for protecting the population.

A Precision Assessment of a Point-of-Care Immunological Analyzer for Swift Progesterone Measurement and Guidance for Determining the Optimal Breeding Time in Bitches.

The measurement of serum progesterone often varies due to different laboratory methodologies and individual canine characteristics. In this investigation, serum progesterone outcomes obtained from a commercial point-of-care immunological analyzer, designed for efficient serum progesterone assessment in bitches, were compared with results derived from chemiluminescent microparticle immunoassay from reference laboratories in Thailand. Our thorough documentation encompassed various parameters: mean, standard deviation, 95% confidence interval, and minimum and maximum serum progesterone concentration values. Additionally, we meticulously recorded the Pearson's correlation coefficient, Lin's concordance correlation coefficient, and the bias correction factor. Interestingly, there was no significant difference (p > 0.05) in the means obtained by the point-of-care immunological analyzer and chemiluminescent microparticle immunoassay. The Pearson's correlation coefficient between the point-of-care immunological analyzer and chemiluminescent microparticle immunoassay stood at 0.957, with Lin's concordance correlation coefficient for point-of-care immunological analyzer recorded as 0.949. Furthermore, the bias correction factor was established at 0.991. This investigation followed established chemiluminescent microparticle immunoassay guidelines, modified to incorporate the mean and 95% confidence interval as criteria for optimal breeding time using the point-of-care immunological analyzer. In conclusion, the commercial point-of-care immunological analyzer emerges as a valuable tool, aiding in precisely determining the optimal timing for natural mating or artificial insemination in bitches.

A Critical Investigation of Sick Euthyroid Syndrome in Chronic Heart Failure Patients: Addressing the Need for Accurate Thyroid Assessment.

Background The body undergoes numerous metabolic changes during severe illness or physiological stress to protect itself by lowering metabolism and reducing overall demands. This evolutionary adaptation dates back to early human development, long before the advent of ICU facilities and advanced treatments. One such protective mechanism is Sick Euthyroid Syndrome (SES), also known as Non-thyroidal Illness Syndrome (NTIS). SES commonly occurs in critically ill patients and is frequently observed in conditions such as heart failure, chronic kidney disease, and severe sepsis. This syndrome is characterized by abnormal thyroid function tests in patients with acute or chronic systemic illnesses who do not have intrinsic thyroid disease. Typically, these patients exhibit low serum levels of triiodothyronine (T3), normal or low levels of thyroxine (T4), and normal or low thyroid-stimulating hormone (TSH) levels. SES is believed to be an adaptive response to illness, aimed at reducing the body's metabolic rate and conserving energy during severe physiological stress. This original article delves into SES's prevalence and clinical impact in these settings. Materials and methods The study aims to determine the prevalence of SES in patients with long-standing heart failure, elucidate the relationship between thyroid function and heart failure severity, and assess its impact on various hematological and clinical parameters. This observational, cross-sectional study was conducted at Dr. D. Y. Patil Medical College, Hospital and Research Centre, Pune, India, a 2011-bed hospital, over one and a half years. This study included 70 patients with chronic heart failure, aged 18 years and above, defined by a left ventricular ejection fraction of 40% or less and a Boston criteria score of 8 or more. Patients were excluded if they had a history of thyroid dysfunction, clinical sepsis, or were taking thyroid-affecting drugs.  Results The study provides important insights into the prevalence and impact of SES in long-standing heart failure patients. It found that a significant 44.29% of these patients exhibited low T3 levels, highlighting the substantial occurrence of SES in this population. Additionally, the study revealed a negative correlation between N-terminal pro-b-type natriuretic peptide (NT-proBNP) levels, Boston score, and total T3, suggesting that as indicators of heart failure severity worsen, total T3 levels may decrease further. Another key finding is the high prevalence of anemia among heart failure patients, with a notable gender disparity: 92.11% of male patients were affected compared to 50% of female patients.  Conclusion The study concluded that SES is significantly prevalent among long-standing heart failure patients, further indicating that thyroid suppression increases with the severity of heart failure. Recognizing SES can guide tailored treatments, prompting intensive monitoring and optimized heart failure management. Additionally, the study found a high prevalence of anemia, particularly among male patients, highlighting the need for gender-specific considerations in managing heart failure. These findings underscore the importance of routine thyroid function assessments and regular monitoring of anemia in heart failure patients. Future research should focus on improving clinical outcomes through comprehensive management of both thyroid function and anemia in these patients.

Development of an automated chemiluminescent immunoassay for cancer antigen 72-4 and the evaluation of its analytical performance.

Objectives: Cancer antigen (CA) 72-4 assay is widely used for monitoring gastric and ovarian cancers. The antigen is a mucin-like, tumor-associated glycoprotein known as TAG-72. It has been identified and characterized using two different monoclonal antibodies, CC49 and B72.3, which recognize its glycochain epitopes, Galß(1-3) sialyl-Tn and sialyl-Tn antigens, respectively. This study describes the quantitative analytical performance of a newly developed CA 72-4 assay, ARCHITECT CA 72-4. Design: and Methods: The ARCHITECT CA 72-4 assay was developed using the ARCHITECT i2000SRs and three ARCHITECT i1000SRs. The assay performance was evaluated based on guidance from CLSI (Clinical and Laboratory Standards Institute) and correlation against Elecsys CA 72-4. Results: In the total precision study, the minimum coefficient of variation (CV) for Control/Panel samples over 4 U/mL was 1.1%. The measuring interval was from 0.95 to 200 U/mL with good linearity; and limits of blank (LoB), detection (LoD), and quantitation (LoQ) were 0.09, 0.18, and 0.95 U/mL, respectively. High dose hook effect; differences among specimen tube types; and interference of common drugs, potential cross-reactants, and endogenous substances were not observed. Significantly, this assay has high biotin tolerance at 4875 mg/mL and correlates well with the Elecys CA 72-4 assay (correlation coefficient: 0.95). Conclusions: ARCHITECT CA 72-4 is a highly sensitive and precise assay for CA 72-4 measurement in human sera and plasma.

Development of a new automated IL-6 immunoassay.

OBJECTIVES: Quantitative detection of interleukin-6 (IL-6) in serum and plasma can help monitor immune responses and the development of acute inflammation to guide patient management. We developed an IL-6 immunoassay for use with the automated ARCHITECT system for detecting an increase in the inflammatory response. METHODS: Immunized mouse sera were tested and selected B-cells were harvested for fusion with myeloma cells. A panel of monoclonal antibodies were produced, from which capture and detection monoclonal antibodies for the prototype IL-6 immunoassay were selected and screened on the ARCHITECT instrument. The antibody pair that most effectively captured and detected IL-6 was selected to develop a prototype IL-6 immunoassay. Calibrator and panel preparations using an internal recombinant IL-6 standard were compared to serum panels prepared with the IL-6 International Standard 89/548. Assay specificity and spike recovery were determined, and assay sensitivity was compared with the Roche EUA Elecsys IL-6 assay run on the cobas analyzer. RESULTS: Twenty-one antibodies in 441 antibody pairs were screened. The prototype IL-6 assay showed high sensitivity with an estimated limit of detection of 0.317 pg/mL and limit of quantitation of <1.27. Spike recovery was 90%-110% in serum and plasma. The internal recombinant human IL-6 calibrator showed excellent stability for 63 days at 2-8 °C. The prototype IL-6 immunoassay was specific for IL-6, exhibited no cross reactivity to related cytokines and interleukins, and was 10-fold more sensitive than the Elecsys IL-6 assay. CONCLUSIONS: The prototype ARCHITECT IL-6 automated immunoassay is a reliable and robust method for the quantitative determination of IL-6 in human serum and plasma.

Inactive SARS-CoV-2 vaccine generates high antibody responses in healthcare workers with and without prior infection.

BACKGROUND AND OBJECTIVES: Healthcare workers (HCWs) were among the first groups to be vaccinated in Turkey. The data to be obtained by the vaccination of HCWs would guide wide spread vaccination programs. MATERIALS AND METHODS: The study included 330 HCWs working at Istanbul University-Cerrahpasa, Cerrahpasa Medical Faculty Hospital and vaccinated with inactive CoronaVac (Sinovac Life Sciences, China) SARS-CoV-2 vaccine in two doses (28 days apart). Anti-Spike /RBD IgG levels were measured 14 days after the first dose and 28 days after the second dose. Chemiluminescent microparticle immunoassay (CMIA) (ARCHITECT IgG II Quant test, Abbott, USA), which is 100% compatible with plaque reduction neutralization test (PRNT), was used. RESULTS: Of the participants, 211 (63.9%) were female, 119 (36.1%) were male, and mean age was 39.6 ± 7.7 years. In those without prior COVID-19 history; (n = 255) antibody positivity was detected as 48.2% (95% CI: 42.1-54.3) 14 days after the first dose of vaccine, and 99.2% (95% CI: 98.1-100) at day 28 after the second dose. Antibody titers were significantly lower in patients with hypertension (p = 0.011). In those with prior history of COVID-19 (n = 75); both the antibody positivity rates after the first vaccine (48.2% vs 100%, p = 0.000) and the anti-spike/RBD antibody levels after the second vaccine (with a ≥ 1050 AU/mL titer equivalent to PRNT 1/80 dilution) was significant than infection-naive group (25.9% vs. 54.7%, p = 0.000). Antibody positivity after two doses of vaccination for all study group was 99.4% (95% CI: 98.6-100). CONCLUSIONS: Two doses CoronaVac produce effective humoral immunity in HCWs. Antibody response is significantly higher in those with prior history of COVID-19 than infection-naive group. Given no significant benefit of the second dose, a single shot of vaccination may be sufficient for those with prior history of COVID-19. Monitoring humoral and cellular immune responses, considering new variants, is required to validate this approach.

A suspected case of falsely low digoxin and vancomycin concentrations caused by free kappa light chains with PETINIA method.

Vancomycin and digoxin are associated with potential toxicity and serum concentrations need to be monitored in certain patients. Previous reports suggested IgM paraproteins could interfere with vancomycin assays, and no paraprotein interference has been reported with digoxin assays. Here we present a suspected case of free-kappa light chains-mediated falsely low digoxin and vancomycin concentrations with Abbott particle-enhanced turbidimetric inhibition immunoassay (PETINIA) method. A 53-year-old patient received multiple doses of vancomycin and digoxin intravenously, but trough vancomycin and random digoxin concentrations repeatedly measured as <1.1 µg/mL and <0.2 ng/mL respectively with Abbott PETINIA method. Results from alternative methods showed concentrations reaching toxic levels and administration of the drugs was immediately terminated. A significantly elevated level of free-kappa light chains, possibly in polymeric form as suggested by protein electrophoresis result, was suspected to be the cause of falsely low results. During the laboratory investigation, absorbance curves revealed increased agglutination in the patient's samples in the latter part of the reaction, suggesting interfering substances led to production of turbidity after reagents were added. Protein-free filtration partially recovered the drugs with Abbott PETINIA. When drug concentrations do not correlate with clinical judgment, clinicians and pharmacists should consult clinical laboratories for investigation of potential interfering substances.

Primary adrenal mature teratoma: A rare case report and review of literature.

Introduction: Teratoma is a germ cell tumor, deriving from totipotent cells. Teratomas usually occur in gonads and are rarely extra-gonadal. The adrenal location is exceptional. Only few cases of primary adrenal teratomas have been reported in literature, mainly in young patients. Case report: We report the case of a 56-year-old female patient who presented with pyrosis, dyspepsia and abdominal pain that was evolving for 5 months. The abdominal computed tomography revealed a voluminous mass of the right adrenal gland and the hormonal evaluation was normal. The patient underwent an open transperitoneal adrenalectomy and the histopathological examination of the specimen confirmed the diagnosis of mature teratoma. Discussion: Adrenal teratomas are commonly asymptomatic and their diagnosis depends mostly on radiologic findings. Malignant transformation is very rare. Surgical excision is the mainstay of treatment with a good prognosis. Conclusion: Open surgery should always be considered in large and adhering teratoma tumors of the adrenal gland. Some pathologic features and tumorigenesis of adrenal teratomas are not entirely elucidated, thus the importance of larger studies in order to comprehend this pathological entity.

Monitoring of anti-SARS-CoV-2 IgG antibody immune responses in two cohorts of Hungarian healthcare workers following infection or immunization / Az anti-SARS-CoV-2-IgG-antitest-immunválaszok monitorozása magyarországi egészségügyi dolgozók két kohorszában fertozést, illetve immunizálást követoen.

Összefoglaló. Bevezetés: A SARS-CoV-2-fertozések és az anti-SARS-CoV-2-vakcinák által kiváltott immunvédelem tartóssága, nagysága és különbségeinek háttere nem teljesen tisztázott, az oltási protokollok optimális idozítése vitatott. Célkituzés: A humorális immunválaszok nagyságát, idobeli változását, a reinfekciók gyakoriságát, demográfiai és klinikai paraméterekkel való összefüggését vizsgáltuk magyarországi egészségügyi dolgozóknál. Módszerek: Megyei egyetemi oktató kórházunkban prospektív, longitudinális vizsgálatot végeztünk egészségügyi dolgozók két csoportjában. 1. kohorsz: SARS-CoV-2-fertozésen átesett, oltatlan 42 dolgozó (no: 100%) antinukleokapszid-IgG-szintjét mértük 8 hónapon keresztül (2020. június-2021. február). Az immunválasznak a változását és az életkorral, a krónikus betegségekkel, a vércsoporttal és a tünetek súlyosságával való összefüggését vizsgáltuk. 2. kohorsz: két dózis mRNS-vakcinával (Pfizer-BioNTech) végzett immunizálást követoen, fertozésnaiv 49 dolgozó (no: 73%) anti-spike-RBD-protein-IgG-szintjét monitoroztuk 8 hónapig (2020. december-2021. augusztus). Medián analízis, lineáris regresszió, ANCOVA, Kruskal-Wallis- és Skillings-Mack-teszt-elemzéseket végeztünk. Eredmények: 1. kohorsz: az IgG-szintek átlagosan a betegség 4-es súlyossági kategóriájában voltak a legmagasabbak, a negatív tartományba csökkenés medián ideje 6 hónap volt. 2. kohorsz: a második vakcina hatására az IgG-szint a 25-szörösére nott, majd 210 nap után a csúcsszint 6%-ra csökkent. Az ellenanyagtiter negatív összefüggést mutatott az idosebb életkorral és a férfinemmel. Tünetmentes (újra)fertozodést valószínusítettünk a fertozésen átesettek 17%-ánál és az immunizált kohorsz 14%-ánál. Az érintettek magas kockázatú osztályokon dolgoztak. Következtetés: 6 hónap után mind a fertozésen átesettek, mind az immunizáltak jelentosen csökkeno IgG-védelmet mutattak. A (re)infekciók átlagosan 15%-ban, tünetmentesen zajlottak. Az eredmények megerosítik az oltás hatékonyságát a betegség megelozésében, a harmadik emlékezteto vakcina fontosságát 6 hónap után és az anti-SARS-CoV-2-IgG-monitorozás potenciális értékét. Orv Hetil. 2022; 163(12): 455-462. INTRODUCTION: The length, level and variation of immune responses to infection with SARS-CoV-2 or following anti-SARS-CoV-2 vaccination remains unclear, optimal (re)vaccination protocols remain debated. OBJECTIVE: We investigated the magnitude of humoral immune responses, their over-time changes, the frequency of (re)infections and the association with demographic and clinical parameters in Hungarian healthcare workers. METHODS: We conducted a prospective, longitudinal study in two groups of healthcare workers of a public, county-level teaching hospital. Cohort 1: The anti-nucleocapsid IgG levels of 42 workers (female: 100%) were followed up over 8 months after SARS-CoV-2 infection (June 2020-February 2021). The change in humoral immune response and its associations with age, existing chronic conditions, blood type and severity of symptoms were investigated. Cohort 2: The anti-spike-RBD protein IgG levels of 49 workers (female: 73%) with no prior COVID-19 infection were monitored over 8 months (December 2020-August 2021) following immunisation with two doses of mRNA vaccine (Pfizer-BioNTech). Analyses included median analysis, linear regression, ANCOVA, Kruskal-Wallis and Skilling-Mack tests. RESULTS: Cohort 1: IgG levels were on average the highest among those in illness severity category 4, the median time of IgG level reduction below the positive test cut-off was 6 months. Cohort 2: The IgG levels increased 25-fold between the first and second immunisations, but decreased to 6% of the peak level after 210 days. They showed an overall negative association with older age and male sex. The suspected levels of (re)infections were 17% and 14% within the infected and the immunised cohorts, respectively, all symptomless. Those affected all worked on high-risk wards. CONCLUSION: Both the infected and the immunised cohorts showed significantly declining IgG protections beyond 6 months. The average observed rate of (re)infections was 15%, all asymptomatic. Our findings are confirmative of the effectiveness of vaccination to prevent illness, the importance of booster vaccination due to declining humoral immune protection beyond 6 months, and the potential value of anti-SARS-CoV-2 IgG monitoring. Orv Hetil. 2022; 163(12): 455-462.

Human hepatocyte-enriched miRNA-192-3p promotes HBV replication through inhibiting Akt/mTOR signalling by targeting ZNF143 in hepatic cell lines.

Previous studies have revealed multiple tissue- or cell-specific or enriched miRNA profiles. However, miRNA profiles enriched in hepatic cell types and their effect on HBV replication have not been well elucidated. In this study, primary human hepatocytes (PHHs), Kupffer cells (KCs), liver sinusoidal endothelial cells (LSECs), and hepatic stellate cells (HSCs) were prepared from liver specimens of non-HBV-infected patients. Four hepatic cell type-enriched miRNA profiles were identified from purified liver cells miRNA microarray assay. The results revealed that 12 miRNAs, including miR-122-5p and miR-192-3p were PHH-enriched; 9 miRNAs, including miR-142-5p and miR-155-5p were KC-enriched; 6 miRNAs, including miR-126-3p and miR-222-3p were LSEC-enriched; and 14 miRNAs, including miR-214-3p and miR-199a-3p were HSC-enriched. By testing the effect of 11 PHH-enriched miRNAs on HBV production, we observed that miR-192-3p had the greatest pro-virus effect in hepatic cell lines. Moreover, we further found that miR-192-3p promoted HBV replication and gene expression through inhibiting Akt/mTOR signalling by direct targeting of ZNF143 in HepG2.2.15 cells. Additionally, the serum and hepatic miR-192-3p expression levels were significantly higher in chronic hepatitis B patients than in healthy controls and serum miR-192-3p positively correlated with the serum levels of HBV DNA and HBsAg. Collectively, we identified miRNA profiles enriched in four hepatic cell types and revealed that PHH-enriched miR-192-3p promoted HBV replication through inhibiting Akt/mTOR signalling by direct targeting of ZNF143 in hepatic cell lines. Our study provides a specific perspective for the role of hepatic cell type-enriched miRNA in interaction with viral replication and various liver pathogenesis.