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1.
Open Access Macedonian Journal of Medical Sciences ; Part A. 11:122-126, 2023.
Article in English | EMBASE | ID: covidwho-20237213

ABSTRACT

BACKGROUND: The coronavirus disease 2019 (COVID-2019) causes the severe contagious acute respiratory syndrome. Therefore, massive vaccination campaign is mandatory to control the spread. Sputnik COVID-19 vaccines induce immunity through different mechanisms involving antibody response that bind to the spike protein to neutralize the viral entry into the cells. AIM: This study aims to compare the titers of specific antibodies in the pre-and post-vaccination sera in the vaccinated Egyptian population to evaluate the efficacy of the sputnik vaccine. METHOD(S): Samples were collected from 205 adult volunteers receiving the Sputnik vaccine in the Reference Laboratory of Egyptian University Hospitals. Samples were collected before vaccination and within 1, 2, or 3 months after receiving two doses of Sputnik SARS-CoV-2 vaccines from August to October 2021, serum samples collected were tested by quantitative chemiluminescent immunoassay using (Mindray CL-960i chemiluminescence analyzer, India) at the Reference laboratory of Egyptian University Hospitals for neutralizing antibodies, anti-spike antibodies, and total antibody levels before and after vaccination. RESULT(S): The results of the 205 paired samples illustrated that there was a statistically significant difference between pre-and post-vaccination antibody levels with a p-value of (< 0.001) indicating that the vaccine produced significantly high levels of antibodies. CONCLUSION(S): COVID-19 Sputnik vaccines induce immunity through an antibody response that binds to the virus to neutralize its entry into cells. Our study showed a significant increase in the measured post-vaccination levels of the three antibodies among the enrolled volunteers compared to the basal pre-vaccination level and thus sputnik vaccine protects against SARS-CoV-2 infections.Copyright © 2023 Ghada Ismail, Dalia Abdelhamid, Marwa Salah Mostafa, Noha Alaa Eldin Fahim, Ahmed Elshafei, Hossam Abdelghaffar, Nashwa Naguib, Omnia Taher, Menna Asker.

2.
Cancer Research Conference: American Association for Cancer Research Annual Meeting, ACCR ; 83(7 Supplement), 2023.
Article in English | EMBASE | ID: covidwho-20235730

ABSTRACT

Objective: During the COVID-19 pandemic, cancer patients had restricted access to standard of care tissue biopsy. Liquid biopsy assays using next generation sequencing technology provides a less invasive method for determining circulating tumour mutations (ctDNA) associated with targeted treatments or prognosis. As part of deploying technology to help cancer patients obtain molecular testing, a clinical program was initiated to offer liquid biopsy testing for Canadian patients with advanced or metastatic breast cancer. Method(s): Blood was drawn in two 10 mL StreckTM DNA BCTs and sent to the CAP/CLIA/DAP accredited Imagia Canexia Health laboratory for testing using the clinically validated Follow ItTM liquid biopsy assay. Plasma was isolated using a double spin protocol and plasma cell-free DNA (cfDNA) extracted using an optimized Promega Maxwell RSC method. Extracted cfDNA was amplified using the multiplex amplicon-based hotspot 30 or 38 gene panel and sequenced. An inhouse developed bioinformatics pipeline and reporting platform were used to identify pathogenic single nucleotide variants (SNVs), indels (insertions and deletions), and gene amplification. Included in the panel are genes associated with metastatic breast cancer: AKT1, BRAF, ERBB2, ESR1, KRAS, PIK3CA, TP53. Result(s): To identify biomarkers, 1214 metastatic or advanced breast cancer patient cfDNA samples were tested. There were 15 cases sent for repeat testing. We reported 48% of samples harboring pathogenic ctDNA mutations in TP53 (22%), PIK3CA (19%), ESR1 (18%), AKT1 (2%), ERBB2 (1.5%). Co-occurring variants were identified in samples with ESR1/PIK3CA as well as TP53/PIK3CA (both p-values <0.001). Interestingly, 29% of samples with mutated ESR1 harbored >= 2 ESR1 ctDNA mutations. In 56% of cases, previous molecular testing indicated the cancer subtype as hormone receptor (ER, PR) positive with/without HER2 negative status. In this specific subgroup, 49% harbored ctDNA mutations with 63% of those being PIK3CA and/or ESR1 mutations. Conclusion(s): A population of Canadian women with metastatic breast cancer were tested using a liquid biopsy gene panel during the COVID-19 pandemic for identification of biomarkers for targeted therapeutic options. Over 50% of the samples were identified as hormone positive, with greater than 60% harboring PIK3CA and ESR1 ctDNA mutations. Studies have shown that metastatic PIK3CA mutated ER-positive/HER2-negative tumors are predictive to respond to alpelisib therapy and have FDA and Health Canada approval. Additionally, ESR1 mutations are associated with acquired resistance to antiestrogen therapies, and interestingly we identified 29% of ESR1 mutated samples with multiple mutations possibly indicating resistance subclones. In future studies, longitudinal monitoring for presence of multiple targetable and resistance mutations could be utilized to predict or improve clinical management.

3.
Germs ; 12(4):507-518, 2022.
Article in English | EMBASE | ID: covidwho-20234801

ABSTRACT

Introduction In this study, we aimed to monitor anti-spike and anti-nucleocapsid antibodies positivity in healthcare workers (HCWs) vaccinated with two doses of inactivated CoronaVac (Sinovac, China) vaccine. Methods Overall, 242 volunteer HCWs were included. Of the participants, 193 were HCWs without history of prior documented COVID-19 (Group 1), while 49 had history of prior documented COVID-19 before vaccination (Group 2). The participants were followed up for SARS-CoV-2 antibodies positivity at four different blood sampling time points (immediately before the second vaccine dose and at the 1st, 3rd months and 141-150 days after the second dose). We investigated the serum IgG class antibodies against SARS-CoV-2 RBD region and IgG class antibodies against SARS-CoV-2 nucleocapsid antigen by chemiluminescent microparticle immunoassay (CMIA) method using commercial kits. Results We found positive serum anti-RBD IgG antibody in 76.4% of the participants (71% in Group 1;98% in Group 2) 28 days after the first dose. When the antibody levels of the groups were compared at the four blood sampling time points, Group 2 anti-RBD IgG levels were found to be significantly higher than those in Group 1 at all follow-up time points. Although anti-RBD IgG positivity persisted in 95.6% of all participants in the last blood sampling time point, a significant decrease was observed in antibody levels compared to the previous blood sampling time point. Anti-nucleocapsid IgG antibody was positive in 12 (6.2%) of participants in Group 1 and 32 (65.3%) in Group 2 at day 28 after the first dose. At the fourth blood sampling time point, anti-nucleocapsid antibodies were found to be positive in a total of 20 (9.7%) subjects, 10 (6.1%) in Group 1 and 10 (23.8%) in Group 2. Conclusions In this study, it was determined that serum antibody levels decreased in both groups after the third month after the second dose in HCWs vaccinated with CoronaVac vaccine.Copyright © GERMS 2022.

4.
International Journal of Infectious Diseases ; 130(Supplement 2):S100, 2023.
Article in English | EMBASE | ID: covidwho-2322005

ABSTRACT

Intro: Different vaccines against COVID-19 have been approved by the World Health Organization (WHO) at different stages, however, limited data is available on long-term kinetics of antibodies induced by vaccines. This study was performed to investigate the persistence and dynamicity of BBV-152 (Covaxin)- and AZD1222 (Covishield)-induced immunoglobulin-G (IgG) antibodies over the year and neutralizing antibodies' status after the one-month post booster dose. Method(s): This 52-week longitudinal cohort study documented antibody persistence and neutralizing antibody status among 278 health-care workers (HCWs) from four different healthcare and research facilities in Odisha, enrolled in January 2021 and continued until March 2022. An automated chemiluminescence immune assay (CLIA) platform from Abbott Diagnostics was used to quantify IgG antibodies against SARS-CoV-2's spike receptor-binding domain (RBD) and a surrogate virus neutralization test (sVNT) was performed by enzyme-linked immunosorbent assay (ELISA). If any participants developed any symptoms of COVID-19, nasopharyngeal swabs were collected and sent to ICMR- RMRC, Bhubaneswar for RT-PCR confirmation. Finding(s): Among the 243 participants, 119 HCWs (48.97%) were Covaxin recipients and the remaining 124 (51.02%) were Covishield recipients. During the seven follow- ups, 104 participants (42.79%) were identified as vaccine breakthrough cases. In 139 non-infected HCWs, the median antibody titer significantly waned after ten months of double dose, both for Covaxin (342.7 AU/mL at DD1 vs 43.9 AU/mL at DD10) and Covishield (2325.8 AU/mL at DD3 vs 595.2 AU/mL at DD10). No statistically significant differences in antibody titers were observed based on age, gender, comorbidities, and blood groups. The median inhibition activity of sVNT was increased significantly for Covaxin and Covishield booster recipients. Among the booster dose recipients, 24 had breakthrough cases by the Omicron variant. Conclusion(s): Results of this longitudinal cohort study can be used to implement vaccination strategies and could also aid in tracking and designing vaccine mandates to minimize vaccine escape.Copyright © 2023

5.
Methods Mol Biol ; 2663: 487-504, 2023.
Article in English | MEDLINE | ID: covidwho-2323039

ABSTRACT

Thrombotic thrombocytopenic purpura (TTP) is a prothrombotic condition caused by a significant deficiency of the enzyme, ADAMTS13 (a disintegrin and metalloproteinase with a thrombospondin type 1 motif, member 13). In the absence of adequate levels of ADAMTS13 (i.e., in TTP), plasma VWF accumulates, in particular as "ultra-large" VWF multimers, and this leads to pathological platelet aggregation and thrombosis. In addition to TTP, ADAMTS13 may be mildly to moderately reduced in a range of other conditions, including secondary thrombotic microangiopathies (TMA) such as those caused by infections (e.g., hemolytic uremic syndrome (HUS)), liver disease, disseminated intravascular coagulation (DIC), and sepsis, during acute/chronic inflammatory conditions, and sometimes also in COVID-19 (coronavirus disease 2019)). ADAMTS13 can be detected by a variety of techniques, including ELISA (enzyme-linked immunosorbent assay), FRET (fluorescence resonance energy transfer) and by chemiluminescence immunoassay (CLIA). The current report describes a protocol for assessment of ADAMTS13 by CLIA. This protocol reflects a rapid test able to be performed within 35 min on the AcuStar instrument (Werfen/Instrumentation Laboratory), although certain regional approvals may also permit this testing to be performed on a BioFlash instrument from the same manufacturer.


Subject(s)
COVID-19 , Purpura, Thrombotic Thrombocytopenic , Humans , Purpura, Thrombotic Thrombocytopenic/diagnosis , von Willebrand Factor , Luminescence , ADAM Proteins , COVID-19/diagnosis , ADAMTS13 Protein
6.
Allergy: European Journal of Allergy and Clinical Immunology ; 78(Supplement 111):320, 2023.
Article in English | EMBASE | ID: covidwho-2302815

ABSTRACT

Background: In the last two years the pandemic Coronavirus Disease 19 (Covid19), caused by the virus SARS-CoV- 2, described for the first time in Wuhan (China) at the end of 2019, has caused over 359 million cases of infections and 5 million deaths worldwide. To fight this emergency, the pursuit of science has focused on vaccines development against SARS-CoV- 2, including the vaccine BNT162b2. This vaccine contains mRNA translating for SARS-CoV- 2 spike protein wrapped in lipid nanoparticles and its use was approved at the end of 2020. It has been proved that both the BNT162b2 vaccine and the SARS-CoV- 2 infection result in the production of neutralizing antibodies but remains to be clarified the duration of these responses, also versus variants of concern. Method(s): The present study aimed to prospectively analyse and correlate the antibody response and the neutralization capability induced by vaccination with BNT162b2 in a cohort of Sardinian subjects, including a group previously Cov2 infected. Each participant was evaluated for serum SARS-CoV2 Ab IgG RDB, 7 (T1) and 30 (T2) days after the second inoculum of BNT162b2, with chemiluminescent immunoassays (CLIA) and microneutralization assay (MNA) determining the highest serum dilution protecting 90 % of the infected wells. Result(s): All the participants, with or without previous infection, developed a positive antibody response (IgG anti-RBD > 1 AU/ml) within 7 and 30 days from the second vaccine dose and a strong correlation was found between IgG antibody levels and neutralizing activity. A strong difference was observed between the antibody levels of the naive subjects and the ones previously infected, specifically the antibody levels were higher (both at T1 and T2) in the latter group. No significant antibody differences were found for gender and age groups. In addition, there were no significant differences in antibody titre between healthy and immune-mediated subjects. Conclusion(s): In conclusion, this study confirms observed differences in vaccine responses between infection-naive and subjects with history of natural infection, with the presence in the second group of a significantly higher neutralizing and anti-RBD antibody titer. It also demonstrates the strong correlation between anti-RBD antibody titre and neutralizing activity, without significant differences between healthy subjects and subjects with immuno-mediated disease in the short-term. Further follow-up is ongoing in this cohort.

7.
Journal of Liver Transplantation ; 8 (no pagination), 2022.
Article in English | EMBASE | ID: covidwho-2297028
8.
Journal of Laboratory Medicine ; 47(1):41-45, 2023.
Article in English | EMBASE | ID: covidwho-2267163

ABSTRACT

Objectives: Administration of the third dose of severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) vaccine was initiated on December 1, 2021, in Japan. However, data on the long-term effects of this third vaccination remain scarce. Here, we examined the levels of SARS-CoV-2 antibodies in those who received the Pfizer BioNTech (BNT162b2) vaccine, 6 months after the third vaccination. Method(s): Samples from 40 healthy volunteers were used to measure SARS-CoV-2 antibodies with chemiluminescent assays against the receptor-binding domain (RBD) of the virus. Result(s): At 445 days after the first dose of BNT162b2, which is 180 days after the third vaccination, the mean anti-RBD IgG level was 159.4 AU/mL (SD 100.1 AU/mL), which was significantly higher than 144 days after the second vaccination, while mean anti-RBD IgM was baseline level (0.4 C.O.I.). The decline in IgG, 180 days after the third vaccination, was 74.1% (SD 16.1%), which was significantly lower than the 88.6% (SD 4.4%) decline observed 144 days after the second vaccination. Furthermore, we revealed that the reduction in IgG from 14 to 180 days after the third vaccination showed a significant inverse correlation with age, and the higher antibody response in younger participants at 14 days after the third vaccination disappeared at longer time points. Conclusion(s): The long-term durability of the IgG titer was significantly higher following the third vaccination compared with the second vaccination, and the reduction in IgG titer after the third vaccination inversely correlated with age.Copyright © 2022 the author(s), published by De Gruyter, Berlin/Boston.

9.
Indian Journal of Clinical Biochemistry ; 37(Supplement 1):S38, 2022.
Article in English | EMBASE | ID: covidwho-2287332

ABSTRACT

Vitamin D, a fat-soluble vitamin helps the body to absorb and retain calcium and phosphorus.Apart from this primary activity, it exhibits potent antimicrobial and antiinflammatory effects viaimmune-modulatory properties. Vitamin D has shown inhibitory effects on the production of pro-inflammatory cytokines, including TNF-alpha and IL- 6, by various mechanisms, includingdown-regulating viralinduced NFkB activation So, this present study aimed to study the relations of serum calcium, phosphorus and Vitamin D levels in association with severity and mortality in SARSCoV- 2 patients. A total of 150 individuals infected with COVID-19 and 50 healthy individuals were recruited. Cases were divided based on severity (mild, moderate and severe) and outcome (discharged or deceased). Serum Ca, Po4, and ALP were analysed by the direct colourimetric method. Vitamin D was measured using the chemiluminescent immunoassay (CLIA). The median serum calcium, Phosphorus, ALP and vitamin D levels in COVID 19 patients were 8.02 mgldL (IQR, 7.24-8.71), 3.93 mgldL (IQR, 2.97- 4.36), 115 IU/L(IQR, 94-146) and 17.2 ng/mL (IQR, 11.6- 25.9) respectively. On comparing the different severity groups a significant difference was found in Vitamin D (p<0.002), ALP (p<0.00001) and calcium (p<0.0001). The serum calcium levels were significantly positively correlated with Vitamin D levels and negative correlation with the inflammatory markers like IL-6. Similarly, patients with low calcium and vitamin D were found to have a fatal outcome. 838 The multivariable analysis showed that a combination of low calcium and vitamin D with higher age are associated with mortality in COVID-19 patients. Serum calcium and Vitamin D were associated with the clinical severity and prognosis of patients with COVID-19.

10.
Journal of Public Health in Africa ; 13(4) (no pagination), 2022.
Article in English | EMBASE | ID: covidwho-2286900

ABSTRACT

Background. Since the start of the COVID-19 pandemic, Chad has had 7,417 confirmed cases and 193 deaths, one of the lowest in Africa. Objective. This study assessed SARS-CoV-2 immunity in N'Djamena. Methods. In August-October 2021, eleven N'Djamena hospitals col-lected outpatient data and samples. IgG antibodies against SARS-CoV-2 nucleocapsid protein were identified using ELISA. "Bambino Gesu" Laboratory, Rome, Italy, performed external quality control with chemiluminescence assay. Results. 25-34-year-old (35.2%) made up the largest age group at 31.9+/-12.6 years. 56.4% were women, 1.3 women/men. The 7th district had 22.5% and the 1st 22.3%. Housewives and students dominated. Overall seroprevalence was 69.5% (95% CI: 67.7-71.3), females 68.2% (65.8-70.5) and males 71.2% (68.6-73.8). >44-year-old had 73.9% seroprevalence. Under-15s were 57.4% positive. Housewives (70.9%), civil servants (71.5%), and health workers (9.7%) had the highest antibody positivity. N'Djamena's 9th district had 73.1% optimism and the 3rd district had 52.5%. Seroprevalences were highest at Good Samaritan Hospital (75.4%) and National General Referral Hospital (74.7%). Conclusion. Our findings indicate a high circulation of SARS-CoV-2 in N'Djamena, despite low mortality and morbidity after the first two COVID-19 pandemic waves. This high seroprevalence must be considered in Chad's vaccine policy.Copyright © 2022 The Authors and PAGEPRESS PUBLICATIONS.

11.
Reviews and Research in Medical Microbiology ; 33(3):148-159, 2022.
Article in English | EMBASE | ID: covidwho-2260539

ABSTRACT

Rapid diagnosis of coronavirus disease 2019 (COVID-19)-infected patients is urgent in making decisions on public health measures. There are different types of diagnostic tests, such as quantitative PCR assay, antibody, and antigen-based and CRISPR-based tests, which detect genetic materials, viral proteins, or human antibodies in clinical samples. However, the proper test should be highly sensitive, quick, and affordable to address this life-threatening situation. This review article highlights the advantages and disadvantages of each test and compares its different features, such as sensitivity, specificity, and limit of detection to reach a reliable conclusion. Moreover, the FDA- authorized kits and studies' approaches toward these have been compared to provide a better perspective to the researchers.Copyright © 2022 Lippincott Williams and Wilkins. All rights reserved.

12.
Kidney International Reports ; 8(3 Supplement):S457, 2023.
Article in English | EMBASE | ID: covidwho-2250936

ABSTRACT

Introduction: Immunoglobulin (Ig)G antibodies against SARS-CoV-2 are implicated in herd immunity. Humoral response to vaccines in kidney transplant recipients (KTRs) is documented to be sub-optimal. However, the response to non-messenger RNA(mRNA) based vaccines in KTRs is not known Methods: SARS-CoV-2 spike protein IgG antibody response was assessed in KTRs using chemiluminescence immunoassay. Patients were characterized by the number of vaccine doses received and Coronavirus disease 2019 (COVID-19) infection in past. Result(s): Out of 224 KTRs evaluated, 197 (87.94%) had positive S1/S2 IgG anti-SARS-CoV-2 antibodies with a median [IQR] titre of 307.5 AU/ml [91 AU/ml - 400 AU/ml]. High titres (in neutralizing range) were found in 170/224 (75.9%) KTRs. Seropositivity rates after 2 doses of vaccination were significantly higher than unvaccinated KTRs (88.67% vs 66.7%;p = 0.006). After adjusting for cofounders, KTRs with diabetes at the time of vaccination were less likely to develop antibody response (aOR 0.31, 95% confidence interval [CI] - 0.10, 0.90;p = 0.032). Higher eGFR was also an independent predictor of antibody response (aOR 1.04 95% CI - 1.01, 1.08;p = 0.005). KTRs vaccinated with CovishieldTM developed higher antibody response as compared to CovaxinTM (aOR 5.04, 95% CI - 1.56, 16.22;p = 0.007). Conclusion(s): A high rate of seroconversion was seen in KTRs after SARS-CoV-2 vaccination with non mRNA vaccines. The presence of diabetes and decreased eGFR independently predicted lower seroconversion rates. No conflict of interestCopyright © 2023

13.
Anal Bioanal Chem ; 2022 Nov 08.
Article in English | MEDLINE | ID: covidwho-2242236

ABSTRACT

The SARS-CoV-2 pandemic has shown the importance of rapid and comprehensive diagnostic tools. While there are numerous rapid antigen tests available, rapid serological assays for the detection of neutralizing antibodies are and will be needed to determine not only the amount of antibodies formed after infection or vaccination but also their neutralizing potential, preventing the cell entry of SARS-CoV-2. Current active-virus neutralization assays require biosafety level 3 facilities, while virus-free surrogate assays are more versatile in applications, but still take typically several hours until results are available. To overcome these disadvantages, we developed a competitive chemiluminescence immunoassay that enables the detection of neutralizing SARS-CoV-2 antibodies within 7 min. The neutralizing antibodies bind to the viral receptor binding domain (RBD) and inhibit the binding to the human angiotensin-converting enzyme 2 (ACE2) receptor. This competitive binding inhibition test was characterized with a set of 80 samples, which could all be classified correctly. The assay results favorably compare to those obtained with a more time-intensive ELISA-based neutralization test and a commercial surrogate neutralization assay. Our test could further be used to detect individuals with a high total IgG antibody titer, but only a low neutralizing titer, as well as for monitoring neutralizing antibodies after vaccinations. This effective performance in SARS-CoV-2 seromonitoring delineates the potential for the test to be adapted to other diseases in the future.

14.
Journal of Infection and Chemotherapy ; 29(1):15-19, 2023.
Article in English | Scopus | ID: covidwho-2243806

ABSTRACT

Introduction: Anterior nasal sampling (AN) might be more convenient for patients than NP sampling to diagnose coronavirus disease. This study investigated the feasibility of rapid antigen tests for AN sampling, and the factors affecting the test accuracy. Methods: This single-center prospective study evaluated one qualitative (ESP) and two quantitative (LUMI and LUMI-P) rapid antigen tests using AN and NP swabs. Symptomatic patients aged 20 years or older, who were considered eligible for reverse-transcription quantitative polymerase chain reaction using NP samples within 9 days of onset were recruited. Sensitivity, specificity, and positive and negative concordance rates between AN and NP samples were assessed for the rapid antigen tests. We investigated the characteristics that affected the concordance between AN and NP sampling results. Results: A total of 128 cases were recruited, including 28 positive samples and 96 negative samples. The sensitivity and specificity of AN samples using ESP were 0.81 and 1.00, while those of NP samples were 0.94 and 1.00. The sensitivity of AN and NP samples was 0.91 and 0.97, respectively, and specificity was 1.00, for both LUMI and LUMI-P. The positive concordance rates of AN to NP sampling were 0.87, 0.94, and 0.85 for ESP, LUMI, and LUMI-P, respectively. No factor had a significant effect on the concordance between the sampling methods. Conclusions: ESP, LUMI, and LUMI-P showed practical diagnostic accuracy for AN sampling compared to NP sampling. There was no significant factor affecting the concordance between AN and NP sampling for these rapid antigen tests. © 2022 Japanese Society of Chemotherapy and The Japanese Association for Infectious Diseases

15.
International Journal of Laboratory Hematology ; 45(Supplement 1):124, 2023.
Article in English | EMBASE | ID: covidwho-2218847

ABSTRACT

Introduction: Heparin-induced thrombocytopenia (HIT) is a severe adverse reaction to heparin caused by heparin-dependent, platelet-activating anti-plateletfactor 4 (PF4)/heparin antibodies. Vaccine-induced thrombocytopenia and thrombosis (VITT) following Astrazeneca vaccine has been described, associated with IgG anti-PF4 antibodies. Only ELISA immunoassays have been shown to detect anti-PF4 antibodies in these patients. Diagnosis of both HIT and VITT requires confirmation of heparin-dependent, platelets activating antibodies to avoid overdiagnosis and overtreatment Anti-PF4 laboratory assay requests during pandemic are in most of the cases of COVID positive vaccinated subjects, vaccinated healed from COVID19 subjects and COVID negative vaccinated subjects. Aim of our study was to investigate which laboratory test to use in patients with suspected HIT in this particular historical period Methods: Thirty patients with suspected HIT/ VITT were tested with three anti-PF4 immunoassays (HIT Ab Latex Immunoassay-Werfen, Acustar HITIgG CliA-Werfen, HPIA ELISA-Stago): 8 Astrazenca suspected VITT;1 Moderna and 1 Pfizer suspected VITT, 20 suspected HIT (9/20 COVID positive and 11/20 COVID negative patients). In order to confirm immunoassay positivity Platelet Aggregation Test (PAT) functional test was performed in all patients found to be positive for at least one assay. Result(s): 3/8 suspected Astrazenca VITT tested positive only by ELISA assay 2/3 tested positive by PAT (confirmed VITT). Both patients with suspected Pfizer and Moderna VITT tested negative for all immunoassays. 4/9 COVID patients with suspected HIT tested positive only by ELISA assay, only 1 of them tested positive by PAT. 4/11 COVID negative suspected HIT tested positive by all immunoassays performed and 1/11 tested positive only by ELISA and CliA immunoassays, among these 5 subjects only 2 were confirmed HIT by PAT functional assay Conclusion(s): With the only exception of suspected Astrazeneca VITT no superiority of IgG-ELISA over CliA or Latex immunoassay in sensitivity to HIT was observed. No single immunoassay method detected all probable HIT cases;if a single test is negative, a second immunoassay or a platelet activation assay should be considered where there is strong clinical suspicion Interestingly by using Bayesan diagnosis of HIT and CliA conservative negative cut-off < 0.13 U/ml suggested in the literature (Marchetti et al. 2020) all our CliA negative (results < manufacturer's cut-off of 1.0 U/mL) and ELISA positive suspected HIT/VITT were correctly classified as positive by CliA assay.

16.
Hormone Research in Paediatrics ; 95(Supplement 2):131, 2022.
Article in English | EMBASE | ID: covidwho-2214182

ABSTRACT

Background: 17-hydroxyprogesterone (17-OHP) basal levels greater than 2ng/ml has been related to the need to request an ACTH test to dismiss non-classic congenital adrenal hyperplasia. We have seen an increase in baseline levels in 2021 compared to 2020. Objective(s): To determine how many of the tests that were requested due to a high basal 17-OHP value, were positive. Assess the need to modify the cut-off points to request an ACTH test. To determine the possible relationship of the increase in basal levels of 17-OHP with the BMI SDS. Material(s) and Method(s): retrospective descriptive study of all patients who underwent an ACTH test between January 1, 2020 and December 31, 2021, in a region of northern Spain. We analyze: sex, age, Tanner stage, BMI SDS, baseline 17-OHP and ACTH test results and main reason for requesting. The test was considered: diagnostic with a peak of 17-OHP at 60 minutes higher than 15 ng/ ml, and suggestive if 10-15ng/ml. Laboratory method for the determination of 17-OHP: chemiluminescence immunoassay in Maglumi 2000 Analyzer (Snibe-Diagnostic). The trial meets the quality criteria certified by EQAS QCNet (BioRad-Laboratories, Inc, UK). Result(s): 149 ACTH tests were performed, 45 in 2020 and 104 in 2021. 116 women (77.9%). Mean age: 10.1 years (range 2.2-15.9 years). 114 had already started puberty. 21 BMI>2SDS. Main reason for requesting the ACTH test: elevated baseline 17-OHP 38%(20% in 2020, 45% in 2021), accelerated bone age (>2 years)48%, pubarche 9%. Mean baseline 17-OHP was 2.93ng/ ml(+/-3.6ng/ml), median 1.26ng/ml(range 0.1-26ng/ml) in 2020 and 2.67ng/ml(range 0.56-16.2ng/ml) in 2021, difference not statistically significant (p 0.67), but clinically relevant. ACTH test results: mean peak at 60 minutes was 4.98ng/ml(range 0.1-64). The determination of baseline 17-OHP was above 2ng/ml in 61 patients, and above 4ng/ml in 24. But only in 7 cases the test was positive (diagnostic or suggestive), all them with basal 17OHP>4ng/ml. This cut-off point shows a higher specificity with the same sensitivity. Comparing 2020 with 2021, we found no difference in BMI SDS (p 0.27). Relationship between BMI SDS and basal 17OHP was not confirmed (p 0.81). Conclusion(s): Our study confirms a significant increase in the number of ACTH tests performed, as well as in the basal level of 17-OHP in the last year. A baseline value of 17-OHP greater than 4ng/dl seems to be the most appropriate as the main indicator of the need to perform the test.

17.
Open Forum Infectious Diseases ; 9(Supplement 2):S482-S483, 2022.
Article in English | EMBASE | ID: covidwho-2189783

ABSTRACT

Background. Ensovibep is a multi-specific DARPin (designed ankyrin repeat protein) antiviral in clinical development for treatment of COVID-19. In the Phase 2 EMPATHY study, ensovibep demonstrated greater viral load decline versus placebo. Here we report (1) the efficacy of ensovibep in patients with and without anti-SARS-CoV-2 antibodies at baseline and (2) SARS-CoV-2 mutation emergence data with treatment. Methods. Eligible ambulatory patients with >=2 COVID-19 symptoms (onset within 7 days) and positive SARS-CoV-2 rapid antigen test on day of dosing, were randomized (1:1:1:1) to ensovibep (600, 225 or 75 mg) or placebo as single, IV infusion. Chemiluminescent immunoassays were used for antibody detection (SARS-CoV-2 S1/S2 IgG and SARS-CoV-2 IgM). A pre-specified subgroup analysis was performed based on baseline anti-SARS-CoV-2 antibody status. Analysis of changes in viral genome from baseline to post baseline was performed to evaluate treatment-emergent mutations. Results. Of the patients analyzed, 48.5% had anti-SARS-CoV-2 antibodies at baseline. Baseline log10 SARS-CoV-2 viral load (mean +/-SD) was similar across groups [ensovibep (all doses) 6.5 +/-1.5, placebo 6.2 +/-1.5];> 90% were infected with the Delta (B.1.617.2) variant. SARS-CoV-2 viral load reduction up to Day 8 showed similar effects in favor of ensovibep compared with placebo regardless of the presence of anti-SARS-CoV-2 antibodies (Figure 1). Patients in ensovibep 75 mg, 600 mg, and placebo groups had comparable incidences of emergent mutations, with a higher incidence in the 225 mg group. Based on analysis of 70% of the expected viral sequencing data, two mutations in the key binding residues of ensovibep were observed (Y489H and F486L) in a total of three patients treated with ensovibep. These patients either cleared virus by Day 8 or mutations were transient (occurred at a single time point but not later in the course of infection). (Figure Presented) Conclusion. Ensovibep effectively reduces SARS-CoV-2 viral load regardless of the presence of anti-SARS-CoV-2 antibodies at baseline. Furthermore, there were no emerging mutations of concern, indicating that a single dose administration of ensovibep is associated with minimal selective pressure.

18.
Biochimica Clinica ; 46(3):S111-S112, 2022.
Article in English | EMBASE | ID: covidwho-2169703

ABSTRACT

As is known T-cells play a central role in the immunological response [1]. Nowadays new assays are being developed for the indirect quantification of T-cell memory activity [2]. The aim of this work was to demonstrate Interferongamma Release Assay (IGRA) test could be useful for vaccination monitoring. 23 vaccinated healthcare workers were enrolled in the study after 8 months of the Pfizer BioNTech vaccination. The antibody levels were assessed through Chemiluminescence immunoassay. T cells were indirectly analyzed by an ELISA against INFgamma. Lymphocyte subtyping was evaluated. Statistical analyses were processed. The patients were divided into 3 different groups based on S-RBD and ACE-2 antibody levels: the S-RBD and ACE-2 antibodies were significantly lower in Group 1 than in Group 2 (p<0.001). However, T cells revealed no significant difference between Group 1 and Group 2. Group 3 was the negative control. The results supported the actual role of SARS-CoV-2 T cell, expressed after the vaccine administration and persisting at high concentration over time, despite the antibody levels [3] [4]. Consequently, the new IGRA test was revealed to be an immunological screening that offers information on the protection from SARS-CoV-2 and suggests new strategies for doses administration.

19.
Biochimica Clinica ; 46(3):S131, 2022.
Article in English | EMBASE | ID: covidwho-2169653

ABSTRACT

Serum Amyloid A (SAA) is an acute-phase protein mainly produced by the liver in response to pro-inflammatory cytokines. SAA is primarily produced by hepatocytes in response to the inflammatory cytokines tumor necrosis factor-alpha (TNF-alpha), interleukin (IL)-1beta and IL-6. In healthy individuals, plasma SAA level is within 0.10 mg/L. However, under inflammatory conditions, plasma SAA levels can increase exponentially, even reaching 1000 mg/L or more in some cases. Human SAA expression is upregulated during the acute phase of various viral infections, including cytomegalovirus, herpes simplex virus, measles virus, mumps virus, rubella virus and varicella-zoster virus, and returns to normal during the convalescent phase of infection. Moreover, can be a useful biomarker to predict COVID-19 patient's severity and prognosis.The aim of this study was to evaluate a new chemiluminescence immunoassay for SAA.All serum samples were measured on Maglumi (Snibe platform) and compared with BN ProSpec (Siemens platform), which is used in the routine of the clinical laboratory of the 'Tor Vergata' University Hospital. Analytical precision, correlation coefficient and linearity were assessed. The precision of CLIA system was evaluated by using the commercial normal and high-quality control materials (IQC) recommended by the manufacturer for evaluating precision of SAA. Precision estimation was performed by evaluating triplicate measurements of aliquots of the same samples, performed for a total of five non-consecutive days. Precision data correlated with those declared by the manufacturer. The linearity test was performed using a series of serial dilutions (1/1, 1/2, 1/4, 1/8, 1/16, 1/32, 1/64, 1/128, 1/256 and only diluent) with dilution sample by manufacture. The linearity test showed a correlation coefficient equivalent to 0.997. The results from Snibe platform correlated well with those obtained by Siemens platform with a correlation coefficient of 0.974 (p<0.001). This study demonstrated that the new Snibe SAA test has excellent analytical performance and good reliability and can be used in routine analysis.

20.
Biochimica Clinica ; 46(3):S97, 2022.
Article in English | EMBASE | ID: covidwho-2168924

ABSTRACT

Background: Vaccines represent the most effective tool to contain SARS-CoV-2 infection as well as monitoring their effectiveness is essential to assess individual protection against the pathogen. Immunosurveillance by evaluating anti-spike protein receptor-binding domain (S-RBD) antibodies represents a useful tool to estimate the long immunity against Severe Acute Respiratory Syndrome CoronaVirus 2 (SARS-CoV-2) infection. The aim of this study was to evaluate the kinetics of antibody response in vaccine recipients. Method(s): We measured anti-S-RBD IgG levels by indirect chemiluminescence immunoassay on Maglumi 800 (SNIBE, California) in 1013 healthy individuals naive to SARS-CoV2 infection after two and three COVID-19 vaccine doses. Result(s): Median (IQR) anti-S-RBD IgG levels at the first measurement (baseline) were 1206 (522-2601) BAU/mL. Females displayed significantly higher median baseline anti-SRBD IgG levels than males (1407 vs 1091 BAU/mL, p=0.003). No association was found between age and baseline anti-S-RBD IgG levels. Antibody levels gradually decrease to a steady state after four months since the peak, and the decay is independent of age, sex, vaccine doses, and baseline antibodies titer. The third dose induces a high anti-S-RBD IgG reactivity in individuals with previous high response and trigger a moderate-high anti-S-RBD reactivity also in individuals with an initial low-moderate anti-S-RBD IgG response. Thus, a third SARS-CoV-2 vaccine dose is associated with a significant immunological response. Conclusion(s): Monitoring anti-SRBD IgG levels as a correlate of protection is helpful for answering important questions about virus neutralization and immunity against SARS-CoV-2. A third SARSCoV-2 vaccine dose is associated with a significant immunological response. Thus, our results support the efficacy of the vaccine programs and the usefulness of the third dose.

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