Brazilian Laboratories Research Articles

Academic engagement in Brazilian university laboratories

Abstract In this study, we add new evidence to the literature by investigating how individual and organizational attributes relate to the intensity of academic engagement (AE) in university research laboratories. AE relies on the adoption of mechanisms of interaction between academia and the business sector, facilitating knowledge exchange and collaboration for mutual benefits. The individual attributes of laboratory coordinators corresponded to gender, academic rank, and nonacademic experience. Organizational attributes were proxied by the scale of laboratory research resources, their competence to establish links with industry, and perceived institutional support. From a sample of 240 Brazilian university laboratories, we defined a typology to discriminate laboratories in terms of the intensity with which they practice multiple AE activities. We then investigated differences in the mean vectors of variables representing individual and organizational attributes across laboratories with distinct levels of AE. The results indicated that at the individual level, only the gender of the lab heads showed an association with the intensity of AE. Compared to laboratories with minimal involvement in AE activities, the intensely engaged ones showed higher means for the number of permanent researchers, staff size, and capital stock. Among the highly engaged laboratories, we found a predominance of those with competence to prospect and attract industrial partners and those who received sufficient support from the departments to facilitate collaboration with industry. Idiosyncrasies related to the Brazilian context are discussed in light of new insights typical of a developing country.

Relationships between yield, enzymatic activity, and chemical properties across different soil layers and phenological stages of grapevines in southern Brazil

The activities of the enzymes β-glucosidase (GLU) and arylsulfatase (ARYL) in annual crop soils can now be determined in several commercial laboratories in Brazil. The results are used to assess soil quality and recommend management practices. However, this technology is not yet available for vineyards. The objective of this study was to evaluate whether the activities of soil GLU and ARYL enzymes are related to grapevine yield and chemical properties in sandy vineyard soils, and to define the phenological stage of the crop and the soil sampling layer that best characterize these relationships. Soil and plant samples were collected in a commercial Vitis vinifera L. vineyard located in the extreme south of Brazil during the 2022/23. The soil was sampled in three layers (0–0.05, 0–0.10, and 0–0.20 m) and at four phenological stages (bud burst, full flowering, veraison, and post-harvest). The activities of the soil enzymes GLU and ARYL, the nutrient content of the soil and the leaf, the quality of the must, and the relative yield (RY) of the grapevine were evaluated. The RY was related to the activity of these enzymes. The best regression model adjustments were observed after harvest and in the 0–0.10 m layer. Enzyme activity was also significantly related to soil organic C and total N, as well as other soil properties, mainly P, K, Ca, Mg, Zn, and Mn. The activities of GLU and ARYL enzymes were related to grapevine yield and chemical properties in sandy vineyard soils and soil sampling in the 0–0.10 m layer after harvest is recommended.

Real-World Study on Implementation of Genomic Tests for Advanced Lung Adenocarcinoma in Brazil.

Tissue inadequacy and operational challenges may limit lung cancer comprehensive biomarker testing. Here, we describe the initial implementation of a tailored tissue molecular journey at Oncoclínicas Precision Medicine Laboratory in Brazil, which includes fast-track (FT) non-next-generation sequencing (NGS) assays combined with a broad NGS panel. From 2021 to 2023, all nonsquamous lung cancer samples eligible for the patient support program "Lung Mapping Consortium" at Oncoclínicas & Co were evaluated using the FT panel (immunohistochemistry for PD-L1 and anaplastic lymphoma kinase [ALK], polymerase chain reaction for EGFR and BRAF, and fluorescence in situ hybridization for ROS1) plus a broad DNA and RNA sequencing panel of 180 genes (custom ARCHER panel). From 1,272 samples received by the laboratory, 3% had no tissue for any molecular testing, 20% was not eligible for broad NGS panel as per pathologist assessment (tumor purity and quantity), additional 12% did not reach presequencing analytical thresholds (nucleic acid quantity and/or quality), and 3% had postsequencing failure. Most frequent alterations were KRAS mutations (28.4%, KRASG12C 9.7%), EGFR mutations (23.6%, exon20 insertions 2.9%), ALK fusions (6.4%), MET exon 14 skipping (4.4%), ERBB2 mutations (3.4%), ROS1 fusions (3.1%), and BRAFV600E (1.9%). In 35% of the samples, FT non-NGS tests were the only molecular diagnostics: EGFR mutations (14%), ALK fusions (4.4%), ROS1 fusions (1.8%), and BRAFV600E (0.7%). Overall, high PD-L1 expression (≥50%) was found in 12.3%. This study provides data on the molecular epidemiology of lung adenocarcinoma in Brazil, confirming high prevalence of EGFR mutations, ALK fusions, and MET exon 14 skipping alteration. Biomarker detection is largely affected by biospecimen collection and processing, with one third of the patients eligible for non-NGS testing only, which presents reduced coverage and sensitivity for actionable drivers.

B-114 Reference interval (RI) estimation for TSH in adults using an indirect method with unsupervised machine learning tool and R programming language

Abstract Background Thyroid-stimulating hormone (TSH) plays a fundamental role in regulating thyroid hormones T3 (triiodothyronine) and T4 (thyroxine). Understanding TSH test results is essential for effective management of thyroid disorders and depends, among other factors, on establishing specific reference intervals (RI) for the population being served. We aimed to estimate the reference intervals for TSH in the adult population served by a large diagnostic network in Brazil. Methods We employed data mining followed by estimation using an indirect method through the LabRI™ algorithm with parametric, non-parametric, and robust statistical treatments for outlier exclusion, a series of algorithms programmed in R Language. Included were TSH values from outpatients, aged 20 to 60 years, both sexes, from two analytical platforms. Used as a comparative RI recommended by the American Thyroid Association (ATA, 2014). Data were extracted from the Data Lake, from January to December 2022, from two laboratories in Brazil. Exclusion criteria: samples from hospital sources and referral laboratories; pregnant; some laboratory tests correlated with de TSH test; samples from patients using thyroid hormone and medications that may interfere with hormonal function and TSH laboratory analysis. Results After exclusion criteria, the sample totaled 26,909 adults of both sexes. After excluding 2.24% outliers, the estimated RI was 0.67-4.26 mIU/L, with the comparative RI chosen for this study being the one recommended by ATA of 0.40-4.00 mIU/L. The RI obtained in our study was similar to the Canadian RI harmonization study for TSH (0.60-4.48 mIU/L). Conclusions We estimated the RI, harmonized between Roche and Siemens platforms, to be 0.67-4.26 mIU/L. The RI obtained by the LabRI™ tool is equivalent to the Adeli study (2023) and is under discussion by the company's endocrinology committee for whether to use the estimated RI from our internal study or the one recommended by ATA (2014) in reports.

Requesting a Colleague’s Independent Opinion at Work

ABSTRACT This paper proposes a multimodal conversation analytic approach to an epistemic action, requesting an opinion. It shows how it is performed in a professional context in which the production, distribution and verification of knowledge are fundamental and involve the workplace’s ecology and materiality. Contrary to other settings in which opinions have been described, requesting the opinion of a colleague does not mobilize personal subjective thoughts but rather an expert analysis. The study contributes to an approach of epistemic actions that considers not only the formatting of turns but also the multimodal organization of the performed actions and their embeddedness in a context in which knowledge is at stake. The paper is based on video recordings made in a forensic laboratory in Brazil, a work site in which requesting opinions is crucial within the organization of the ongoing activities. Describing the methodical organization of requesting an independent opinion of a peer in this professional setting, the paper also offers a first characterization of teamwork in the forensic laboratory. Data come from 50 h of video recordings, with participants speaking Brazilian Portuguese.

The Landscape of High-Containment Biological Laboratories in Brazil: Current Status and Perspectives

The Landscape of High-Containment Biological Laboratories in Brazil: Current Status and Perspectives

Profile of Laboratories With the Seal of the Clinical Laboratory Accreditation Program (PALC)

Objective: The objective of this study is to analyze clinical analysis laboratories accredited by the quality seal of the Clinical Laboratory Accreditation Program (PALC) in Brazil. Theoretical Framework: It presents the fundamentals of the research, covering the importance of hospital accreditation, PALC regulations, historical evolution of hospital quality, economic impact on health, quality management in clinical laboratories and the usefulness of quality indicators. Method: A documentary study was carried out, with descriptive and quantitative analysis. During the study period, 163 clinical laboratories were detected in the country accredited by the PALC seal. Results and Discussion: In the study, 163 accredited clinical laboratories were identified in Brazil, with only 32 laboratories in the entire northeast region. Where 92% of accredited laboratories are private and 82% are part of large national diagnostic networks. Highlighting the need for greater investment in quality seals, which better certify the quality of the laboratory service provided. Research Implications: The importance of policies and strategies to promote the accreditation of clinical laboratories stands out, especially in the northeast region of Brazil, aiming to improve the quality and reliability of health services provided to the population, in addition to promoting greater security in medical diagnoses. Originality/Value: This study contributes to the literature by highlighting the originality by examining the grographic distribution and the public and private nature of accredited clinical laboratories in Brazil. The relevance and value of this research are evidenced by its crucial role of accreditation in ensuring quality laboratory services, encouraging continuous improvement and patient confidence.

Abstract PO4-26-07: HER2-low prevalence in early breast cancer (BC) across reference laboratories in Brazil: potential impact of immunohistochemistry (IHC) antibody assay sensitivity

Abstract Background: Current definition of HER2-positive and HER2-low BC follows ASCO/CAP guidelines using IHC and/or in situ hybridization (ISH)-based techniques. Although there is no recommendation for a specific antibody assay, the companion diagnostic test for trastuzumab deruxtecan in the USA is Ventana PATHWAY anti-HER2 monoclonal antibody (4B5) run on Benchmark Ultra instrument. In this study, we assessed the prevalence of HER2-low BC in samples stained with Dako anti-HER2 polyclonal antibody (A0485) and analyzed with Autostainer Link 48 Agilent/Dako, commonly used in different large-volume laboratories in Brazil. Methods: Retrospective study of all early-stage BC samples without exposure to neoadjuvant therapy tested for HR plus HER2 status in Oncoclínicas Precision Medicine (Locus Lab) from 2021 to 2023 and analyzed following ASCO/CAP guidelines. Tumors with HER2 IHC score 0 were classified as HER2-zero, whereas tumors with HER2 score 1+ and those with HER2 score 2+ with ISH-negative were classified as HER2-low. To assess the impact of preanalytical factors on HER2-low positivity, we stratified samples as in-house (part of Oncoclínicas network) or external (other institutions’ FFPE preparation conditions not controlled). Results were also validated in independent IRA Lab using the same Dako anti-HER2 polyclonal antibody (A0485) on Autostainer Link 48 Agilent/Dako. Results: Out of 1,638 eligible samples from Locus Lab, 84% were classified as HR-positive/HER2 negative, 12% HER2-positive, and 4% triple-negative. In 1,386 HR-positive/HER2 negative samples, 83% were HER2-zero, and 17% (CI95% 15%-19%) were HER2-low (1+ in 79% and 2+ with ISH negative in 21%). In 70 triple-negative samples, 62% were HER2-zero and 38% (CI95% 27%-51%) were HER2-low (1+ in 44% and 2+ with ISH negative in 56%). HER2-low positivity was 17% in the subset of in-house samples (N=1.090) and 16% in external referrals (P=0.41). In the independent cohort, out of 441 samples from IRA Lab, 80% were HR-positive/HER2 negative, 12% HER2-positive and 8% triple negative. In 352 HR positive/HER2 negative samples, 80% were HER2-zero and 20% (CI95% 16%-25%) were HER2-low. In 36 triple-negative samples, 72% were HER2-zero and 28% (CI95% 14%-45%) were HER2-low. Conclusions: Different from another Brazilian cohort where the prevalence of HER2-low was 50% with monoclonal anti-HER2 antibodies (Reinert et al., SABCS 2020), our data suggests that the reduced proportion of samples with HER2-low status may be related to antibody clone sensitivity and not linked to preanalytical factors or local ancestry/cancer biology. Indeed, a recent Italian cohort also had lower than expected prevalence of HER2-low in surgical specimens using anti-HER2 Dako A0485 assay on Omnis instrument (Rossi et al., Breast Cancer Res Treat 2023). Prior studies have found differential analytical sensitivity of antibodies for HER2-low status, while HER2 score 3+ detection is equivalent (Ruschoff et al., Virchows Archiv 2022). We recommend prospective evaluation of the HER2-low status with companion-diagnostic Ventana 4B5 assay and alternative antibodies and instruments. Citation Format: Leonard M da Silva, Fernanda Orpinelli Rego, Matheus Costa e Silva, Tomas Reinert, Fernanda Koyama, Daniel Bueno da Cunha, Vagner Duarte, Emílio Pereira, Bruno Ferrari, Rodrigo Dienstmann. HER2-low prevalence in early breast cancer (BC) across reference laboratories in Brazil: potential impact of immunohistochemistry (IHC) antibody assay sensitivity [abstract]. In: Proceedings of the 2023 San Antonio Breast Cancer Symposium; 2023 Dec 5-9; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2024;84(9 Suppl):Abstract nr PO4-26-07.

Emergence of zoonotic sporotrichosis in Brazil: a genomic epidemiology study

Zoonotic sporotrichosis is a neglected fungal disease, whereby outbreaks are primarily driven by Sporothrix brasiliensis and linked to cat-to-human transmission. To understand the emergence and spread of sporotrichosis in Brazil, the epicentre of the current epidemic in South America, we aimed to conduct whole-genome sequencing (WGS) to describe the genomic epidemiology. In this genomic epidemiology study, we included Sporothrix spp isolates from sporotrichosis cases from Brazil, Colombia, and the USA. We conducted WGS using Illumina NovaSeq on isolates collected by three laboratories in Brazil from humans and cats with sporotrichosis between 2013 and 2022. All isolates that were confirmed to be Sporothrix genus by internal transcribed spacer or beta-tubulin PCR sequencing were included in this study. We downloaded eight Sporothrix genome sequences from the National Center for Biotechnology Information (six from Brazil, two from Colombia). Three Sporothrix spp genome sequences from the USA were generated by the US Centers for Disease Control and Prevention as part of this study. We did phylogenetic analyses and correlated geographical and temporal case distribution with genotypic features of Sporothrix spp isolates. 72 Sporothrix spp isolates from 55 human and 17 animal sporotrichosis cases were included: 67 (93%) were from Brazil, two (3%) from Colombia, and three (4%) from the USA. Cases spanned from 1999 to 2022. Most (61[85%]) isolates were S brasiliensis, and all were reported from Brazil. Ten (14%) were Sporothrix schenckii and were reported from Brazil, USA, and Colombia. For S schenckii isolates, two distinct clades were observed wherein isolates clustered by geography. For S brasiliensis isolates, five clades separated by more than 100 000 single-nucleotide polymorphisms were observed. Among the five S brasiliensis clades, clades A and C contained isolates from both human and cat cases, and clade A contained isolates from six different states in Brazil. Compared with Sbrasiliensis isolates, larger genetic diversity was observed among S schenckii isolates from animal and human cases within a clade. Our results suggest that the ongoing epidemic driven by S brasiliensis in Brazil represents several, independent emergence events followed by animal-to-animal and animal-to human transmission within and between Brazilian states. These results describe how S brasiliensis can emerge and spread within a country. Coordenação de Aperfeiçoamento de Pessoal de Nível Superior, Brazil; the São Paulo Research Foundation; Productivity in Research fellowships by the National Council for Scientific and Technological Development, and Ministry of Science and Technology of Brazil.

Molecular Absorption Scanning and Assay of Serotonin Reception Inhibitor Antidepressant Drugs

Background:: Spectrophotometric techniques are based on the absorption of electromagnetic energy by the molecules of the substance analyzed and can be applied to drug analysis. Objective:: This study has been proposed to carry out molecular absorption scanning and assay in the ultraviolet region of generic antidepressant drugs of the serotonin reuptake inhibitor class produced by three Brazilian laboratories, addressing the importance of good manufacturing practices, drug quality control, and the use of qualitative and quantitative analytical techniques. Methods:: The physicochemical analyses were performed in a Marte Científica (580 UVP) UV-Vis spectrophotometer, and the result was evaluated according to the acceptance interval described in the Brazilian Pharmacopoeia. Results:: When calculating the dosage of the fluoxetine hydrochloride sample, the value of 90.95% was obtained and, therefore, the drug was approved for the dosage test. The dosage test for the drugs, citalopram hydrobromide and sertraline hydrochloride, was also approved, obtaining values of 102.4% and 98.2%, respectively. Conclusion:: The results presented provide future perspectives regarding the rigor of the need for good drug handling practices, maintenance and qualification of quality control laboratory equipment, and the guarantee of validation of analytical methodologies used in the pharmaceutical industries, contributing to the improvement of the quality of production and pharmaceutical products analysis.

Compulsory Licensing During Pandemic Times: Possible Lessons From the Efavirenz Case

During the Covid-19 pandemic, partnerships between Brazilian laboratories and international companies were impor tant for the local production of the vaccine, due to the necessity to find an effective measure to preserve public health. However, it is necessary to investigate these partnerships beyond their benefits in order to understand their implications. Compulsory licensing consists of an exception to the transfer of technology contracts to guarantee the Brazilian’s population access to the vaccine and possible new drugs developed against Covid-19. Compulsory licensing has already been implemented in Brazil. In 2007, it was used by the federal government to solve the problem of access to efavirenz, an antiretroviral needed in the treatment of HIV/AIDS. Considering the similarities between the epidemics, this article aimed to answer, through empirical analysis, what the efavirenz case reveals about the legaleconomic process of patent exclusivity and the use of compulsory licensing during times of pandemic crises, such as the one of Covid-19. Based on the analysis of the bureaucratic process applied to the Efavirenz case, it was possible to infer that the compulsory license should have been executed by the Brazilian government to guarantee access to medical products during national emergencies.

Captive Maintenance and Venom Extraction of Tityus serrulatus (Brazilian Yellow Scorpion) for Antivenom Production.

Scorpion envenomation is a public health problem in several tropical and subtropical countries. Tityus serrulatus Lutz and Mello, 1922 (Brazilian yellow scorpion) are responsible for approximately 150,000 envenoming cases per year in Brazil, of which 10% require antivenom treatment to reverse life-threatening venom effects. Therefore, thousands of T. serrulatus individuals are maintained under controlled captivity conditions for venom extraction, subsequently used in the production of the national supply of scorpion antivenom. Instituto Butantan is the main antivenom-manufacturing laboratory in Brazil, providing about 70,000 vials of scorpion antivenom for the Brazilian health system. Thus, the husbandry protocols and venom extraction methodologies are key points for the success of large-scale, standardized venom production. The objective of this article is to describe the captivity protocols of T. serrulatus husbandry, encompassing the husbandry routine and the venom extraction procedures, following good manufacturing practices, and ensuring animal welfare. These practices allow for the maintenance of up to 20,000 animals in captivity, with a routine of 3,000 to 5,000 scorpions milked monthly according to antivenom manufacturing demand, achieving an average of 90% of positive extraction.

B-256 Genotype Distribution of the Hepatitis C Virus: 10 Years Data of a Clinical Laboratory in Brazil

Abstract Background Hepatitis C virus (HCV) infection is a significant health problem worldwide and is a major cause of severe liver disease including chronic hepatitis, cirrhosis, and hepatocellular carcinoma. HCV is divided into six genotypes with several sub-genotypes depending on the geographic region, some genotypes are endemic. The purpose of this study was to describe the frequency of the HCV genotype (G) profile by gender and age using a database of a large Brazilian clinical laboratory. Methods This was a retrospective analysis from January 2012 to December 2022 based on molecular data of 3.566 isolated from clinical samples. The automated extraction and purification of nucleic acids were conducted from EDTA plasma samples of individuals suspected of HCV infection. Then, nucleic acid amplification was performed using oligonucleotide probes labeled with fluorescent dyes specific for the qualitative identification of HCV genotypes 1 to 6 and subtypes A and B of genotype 1. Results A total of 3.566 HCV genotype cases were analyzed during the period. The genotype distribution was (2.653/74.4%) G1 (48.1% G1A and 46.4% G1B), (733/ 20.5%) G3, (133/3.7%) G2, (35/0.9%) G4, (11/0.3%) G5 and (1/0.02%) G6. Twelve patients had multiple genotypes over the last ten years. At least one individual changed the genotype over a 3-year evaluation period, and 11 (0.30%) changed it twice. The genotypes G1 and G3 were predominant in all years analyzed. The prevalence of G1 and G3 was 75% and 19.6% among females and 74% and 21.2% among male patients, respectively. The analysis was conducted on individuals aged 6 to 99 years, with an overall average age of 52.6±13 years. The average age among men was 51.1 ± 12 years and among women, it was 54.7 ± 13 years. According to the age groups evaluated, genotype 1 was the most frequent, ranging from 69.3% (21 to 30 years old) to 75.8% (over 60 years old). The main subtype in the over 60 years old group was 1B. The highest frequency of genotype 2 was observed in the group over 60 years old (6.1%). The patient's symptoms, clinical history, or further diagnostic procedures were not accessible. Conclusion The study found that genotypes 1 and 3 were the most prevalent in Brazil over the ten-year period, with genotype 1 being the most frequent among all age groups. Genotype 1B was most prevalent in individuals above 60 years old. The change of genotype per individual was less than 1%, but further attention is required for potential virus tropism.

A-059 Evaluation of Free Thyroxin Levels in Patients with Low Thyroxin Binding Globulin Concentrations: Is There Interference in the FT4 Indirect Method?

Abstract Background Thyroxine-binding globulin (TBG) is the major carrier of thyroid hormones in serum. Variations in the serum concentration of TBG determine proportional variations in the total serum concentrations of T4 and T3, without implying changes in function, as long as the free fraction remains normal. Several common clinical conditions lead to significant alterations in TBG levels, the most important factor behind low levels being genetic defects. As TBG is encoded by a gene located on the X chromosome, the defects are more easily manifested in males. Differences in FT4 concentrations were observed in sera from subjects with TBG abnormalities, when FT4 are measured by different immunoassays, specially at extremes of TBG concentrations. The objective of this study was to evaluate FT4 levels by indirect method in patients with low TBG concentrations. Methods We analyzed blood samples requesting serum TBG, TSH and FT4 levels from patients admitted to a private reference clinical laboratory in Brazil, from January 01, 2017 to December 31, 2022. Anonymized data on laboratory tests was available from a database of the local Laboratory Information System. All the patients included had TSH within the reference range, 0.4–4.3 mUI/mL (ECLIA, Modular, Roche). FT4 was measured by ECLIA (0.7–1.8 ng/dL) and TBG by ICMA, Immulite, Siemens, 13–39 mcg/mL. Results 357 patients with low TBG levels were evaluated, 220 (62%) men, 137 (38%) women; mean age 35 yrs (19 d to 88 yrs). They were divided in six groups according TBG levels (mcg/mL): <3.5–17 (M 15, F2); 3.5 to 6.0 - 26 (M 20, F 6); 6.1 to 8.0 - 30 (M 25, F 5); 8.1 to 9.9 - 59 ( M 32, F 27); 10.0 to 11.9 - 139 (M 78, F 61); 12.0 to 12.9 - 86 (M 50, F 36). Three patients had FT4 levels above 1.8 (1.85; 1.87 and 1.96) ng/mL, all them male, with TBG levels <3.5; 5.79 and 9.53 ng/mL, respectively. Conclusion FT4 Indirect methods are developed taking into account the presence, in the serum, of a thyroxine binding capacity within the normal range, a fact that does not occur in individuals with extreme TBG deficiency. In this study, we found only one slightly elevated FT4 value among 17 patients with TBG values below 3.5 mcg/mL, suggesting little interference of very low TBG levels in this indirect free T4 method.

B-055 Comparison of Sensitivity Between Tissue Anti-Transglutaminase and Anti-Endomysium IgA and IgG in Serological Screening for Celiac Disease in a Large Clinical Laboratory in Brazil

Abstract Background Celiac Disease (CD) is a genetic and autoimmune disorder characterized by permanent intolerance to gluten found in foods, resulting in small intestine injury in children and adults. Anti-tissue transglutaminase (anti-tTG) and anti-endomysium (anti-EM) antibodies are important serological markers of this disease. The aim was to analyze the analytical sensitivity of these markers in a large-scale Clinical Laboratory routine. Methods We employed a pure sample and a series of dilutions in the analysis of the functional sensitivity of each marker (anti-tTG IgA/IgG and anti-EM IgA/IgG) and a database survey of a large Clinical Laboratory in Sao Paulo/Brazil, from January to December 2021, analyzing 60 000 samples with concomitant results of anti-tTG IGA and anti-EM IgA and 7075 samples with results of anti-tTG IgG and anti-EM IgG, from patients with no defined clinic condition. The anti-tTG tests were performed by fluorescence enzyme immunoassay (FEIA) and the anti-EM tests by indirect immunofluorescence (IFI), with an initial dilution of 1:10, following the manufacturer's recommendations. Results The determination of functional sensitivity by checking the maximum dilution of antibody detection showed higher sensitivity of the anti-tTG IgA and IgG test (dilution factor 0.3 for both) compared to the anti-EM IgA and IgG (dilution factor 0.7 for both). Of the 60 000 samples analyzed for IgA markers, 59 083 (98.5%) were negative for both tests. Of the remaining 917 samples (1.5%), 609 (66.4%) were positive for both anti-tTG IgA and anti-EM IgA, 308 (33.6%) were positive only for anti-tTG IgA, and we did not observe any sample anti-tTG IgA negative with anti-EM IgA positive. Of the 7075 samples analyzed for IgG markers, 7032 (99.4%) were negative for both tests. Of the remaining 43 samples (0.6%), 18 (41.9%) were positive for anti-tTG IgG and anti-EM IgG, 14 (32.6%) were positive only for anti-tTG IgG, and 1 (2.3%) were positive only for anti-EM IgG. Conclusion The study showed greater sensitivity of anti-tTG compared to anti-EM for both IgA and IgG, in agreement with the literature data. The anti-tTG IgA test, in individuals without immunoglobulin A (IgA) deficiency, is recommended as the initial test in suspicion of celiac disease by current scientific guidelines on celiac disease. Based on our findings and literature data, in individuals with IgA deficiency, we suggest a screening using two automated IgG markers, tTG IgG and deamidated gliadin IgG (DG IgG). In addition to the lower sensitivity of anti-EM, indirect immunofluorescence (IFI), compared to the fully automated FEIA method, is a more subjective, observer-dependent, and labor-intensive methodology to be used as screening in large-scale Clinical Laboratory routines.

A-060 Thyroid Laboratory Database Assessment Shows that Total Triiodothyronine Significantly Decreases with Age

Abstract Introduction The effects of aging on human thyroid function are well studied in relationship to thyroid stimulating hormone (TSH) and free tiroxine (FT4); TSH increases, while FT4 decreases with age. Although animal studies show that total triiodotironine (TT3) is lower in aged male rats, these changes are still not clear enough in humans. The National Academy of Clinical Biochemistry (NACB) suggests that TT3 levels would be lower in subjects over 60 years (y.), but the number of elderly assessed was too small for a robust statistical analysis. In search on PubMed, crosschecking the terms elderly/age/aging, with triiodothyronine/T3/total T3, there are a few studies available using current assays with a significant number of patients. Therefore, the demonstration of how the TT3 performs with age is still an opened question. Objective The aim of this study was to observe if TT3 changes significantly with age. Methods We, retrospectively, analyzed blood samples requesting serum TT3 of unique patients, who had TSH measured on the same day; and thyroid peroxidase antibodies (TPOAb), thyroglobulin antibodies (TGAb), anti TSH receptor antibodies (TRAb) or TSH receptor-stimulating immunoglobulin (TSI) at any time since patient admission. Subjects were adults ≥20 y., admitted to a private reference clinical laboratory in Brazil, from January 01, 2014 to December 31, 2022, using anonymized data. Serum TT3 was measured in Cobas Roche, by electrochemiluminescence assay, reference interval 70–210 ng/dL. Exclusion criteria were (and/or): tests performed in hospitals, use of drugs that could interfere on TT3 or TSH, TSH ≤ 0.1 mIU/L or ≥10.0 mIU/L, and any positive antibody at any time. The level of statistical significance was P < 0.05. Results Initially, 959 297 records from the database were surveyed. After applying the exclusion criteria, 141 819 subjects were eligible: mean age = 45 y.; median = 42 y., 62% female (F). Statistically significant differences - P < 0.05 for all comparisons between groups - were found by age and gender in TT3 mean values (in ng/dL). Regarding age: from 20 to 59 y. = 120 ± 26.6; 60 to 79 y. = 109 ± 20.7; and 80 y. and over = 97.2 ± 20.0. By gender: F = 120 ± 28.9; male = 114 ± 20.4. It was also observed that, in elderly, while TT3 decreases, TSH increases (results not shown). Comparing TT3 means by age groups, it is 9% lower in subjects aged 60–79 y. and 19% lower in those aged 80 y. and older, compared to younger (20–59 y.) subjects. Conclusion TT3 results is influenced by several factors, including age, gender and laboratory assay. As far as we know, searching in PubMed, this is the largest collection of data on the relationship between TT3 and age. The data obtained showed a statistically significant decrease in TT3 in elderly, and mainly in very elderly, and higher levels in women than in men. This finding draws call attention to the fact that these parameters should taken into account when evaluating TT3 levels.

Fast and cost-effective protocol to produce Paracoccidioides spp. antigens

The existing methods for Paracoccidioides spp. antigen production are problematic in terms of standardization, specificity, stability, repeatability, and reproducibility. To optimize the methodology for Paracoccidioides spp. antigen production and evaluate its applicability in paracoccidioidomycosis immunodiagnosis. The antigens were obtained from Paracoccidioides lutzii isolates (01, 66, and 8334), Paracoccidioides brasiliensis sensu stricto (113), and Paracoccidioides restripiensis (B-339). These fungi were grown at 36 °C ± 1 °C, on modified Fava-Netto agar, according to Freitas et al. (2018). Paracoccidioides lutzii antigens were obtained after , 10, and 20 days of culture, whereas P. brasiliensis and P. restripiensis antigens were obtained after 10 days. Antigens were evaluated in natura, 10 and 20 times concentrated. Antigenic capacity was evaluated using a double immunodiffusion assay against serum samples from patients with paracoccidioidomycosis, histoplasmosis, and aspergillosis, and random blood donors. Cross-reactivity between Paracoccidioides spp. antigens was observed when P. brasiliensis, P. restrepiensis antigens, and P. lutzii antigens were evaluated with the polyclonal antibodies against P. lutzii and P. brasiliensis, respectively. No cross-reactivity was obtained for polyclonal antibodies against Histoplasma capsulatum, Aspergillus fumigatus, and random blood donors. The proposed protocol allowed stable, repeatable, and reproducible genus-specific antigen production at a low cost and in a short cultivation time. The proposed protocol allowed us to obtain genus-specific antigens that can be developed and reproduced in all laboratories in Brazil and South America, where paracoccidioidomycosis is a neglected disease, contributing to an early diagnosis, especially in endemic regions, regardless of the species.

Brazilian Registry of Interventional Cardiology during the COVID-19 Pandemic (RBCI-COVID19).

At the beginning of the COVID-19 pandemic, patients with myocardial infarction (MI) took longer to present to hospitals because of fear of contamination and health care access difficulties. To assess interventional cardiology procedures performed during the COVID-19 pandemic and its implications for MI approach. Prospective registry of 24 cardiac catheterization laboratories in Brazil, with adult patients undergoing interventional cardiology procedures between May 26 and November 30, 2020. The outcomes were cardiovascular (CV) and non-CV complications, death, and MI. Concomitant COVID-19 was confirmed using RT-PCR. Machine learning techniques were used with nonparametric Classification Trees models, and Simple Correspondence Analysis, with R statistical software package. Significance level adopted of 5%. This study included 1282 patients, 435 of whom (33.9%) had MI as follows: ST-segment elevation MI (STEMI), 239 (54.9%); and non-ST-segment elevation MI (NSTEMI), 196 (45.1%). Of the 1282 patients, 29 had CV complications, 47 had non-CV complications, and 31 died. The diagnosis of COVID-19 was confirmed in 77 patients (6%), with 15.58% mortality and non-CV complications in 6.49%. Most patients had significant coronary artery disease (63%), and an intracoronary thrombus was more often found in the presence of STEMI (3.4%) and COVID-19 (4%). A door-to-table time longer than 12 hours in NSTEMI was associated with 30.8% of complications, 25% in COVID-19 patients. All deaths were preceded by CV or non-CV complications. The presence of COVID-19 was associated with death and non-fatal complications of patients undergoing interventional cardiology procedures during the pandemic.

The Influence of the Omicron Variant on RNA Extraction and RT-qPCR Detection of SARS-CoV-2 in a Laboratory in Brazil.

The emergence of SARS-CoV-2 variants can affect their detection via RT-qPCR. The Omicron variant has a greater affinity for the upper respiratory system and causes clinical conditions similar to bronchitis, as opposed to the pneumonitis-like conditions caused by other SARS-CoV-2 variants. This characteristic increases the viscosity of clinical samples collected for diagnosis. Coinciding with the arrival of the Omicron variant, we observed a failure in control gene expression in our laboratory. In this report, we have optimized a rapid nucleic acid extraction step to restore gene expression and detect the presence of the SARS-CoV-2 virus. We reevaluated 3000 samples, compared variant types occurring in different time periods, and confirmed that the presence of the Omicron variant was responsible for changes observed in the characteristics of these clinical samples. For samples with large amounts of mucus, such as those containing the Omicron variant, a modification to the nucleic acid extraction step was sufficient to restore the quality of RT-qPCR results.

Constant testing for Pestivirus in cell lines reveals different routes of contamination.

Pestivirus can contaminate cell cultures and sera and cause serious problems that evolve the integrity of studies, confidence in diagnostic results, and safety of human and animal vaccines. Contaminations by Pestivirus and other viruses may occur at any time and regular assays of monitoring in cell cultures and your supplies are necessary. This study aimed to analyze the phylogeny of Pestivirus detected from cell cultures, calf serum, and standard strains of three laboratories in Brazil that carry out frequent tests for the monitoring of cellular contaminations. These samples were submitted to phylogenetic analysis to understand the genetic relationship between contaminants occurring in these facilities. As result, the Pestivirus found in samples were Bovine viral diarrhea virus (BVDV-1 and BVDV-2), Hobi-like viruses (often named BVDV-3), and Classical swine fever virus (CSFV), and the phylogenetic analysis help us to infer at three possible routes of contamination in this work.