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1.
J Bras Pneumol ; 50(4): e20240165, 2024.
Artigo em Inglês | MEDLINE | ID: mdl-39356913

RESUMO

OBJECTIVE: To establish the accuracy of frozen section examination in identifying tumor spread through air spaces (STAS), as well as to propose a reproducible technical methodology for frozen section analysis. We also aim to propose a method to be incorporated into the decision making about the need for conversion to lobectomy during sublobar resection. METHODS: This was a nonrandomized prospective study of 38 patients with lung cancer who underwent surgical resection. The findings regarding STAS in the frozen section were compared with the definitive histopathological study of paraffin-embedded sections. We calculated a confusion matrix to obtain the positive predictive value (PPV), negative predictive value (NPV), sensitivity, specificity and accuracy. RESULTS: The intraoperative frozen section analysis identified 7 STAS-positive cases that were also positive in the histopathological examination, as well as 3 STAS-negative cases that were positive in the in the histopathological examination. Therefore, frozen section analysis was determined to have a sensitivity of 70%, specificity of 100%, PPV of 100%, NPV of 90.3%, and accuracy of 92% for identifying STAS. CONCLUSIONS: Frozen section analysis is capable of identifying STAS during resection in patients with lung cancer. The PPV, NPV, sensitivity, and specificity showed that the technique proposed could be incorporated at other centers and would allow advances directly linked to prognosis. In addition, given the high accuracy of the technique, it could inform intraoperative decisions regarding sublobar versus lobar resection.


Assuntos
Secções Congeladas , Neoplasias Pulmonares , Sensibilidade e Especificidade , Humanos , Neoplasias Pulmonares/patologia , Neoplasias Pulmonares/cirurgia , Estudos Prospectivos , Masculino , Feminino , Pessoa de Meia-Idade , Idoso , Reprodutibilidade dos Testes , Período Intraoperatório , Valor Preditivo dos Testes , Idoso de 80 Anos ou mais , Invasividade Neoplásica , Adulto , Cuidados Intraoperatórios/métodos , Pneumonectomia/métodos
2.
Rev Assoc Med Bras (1992) ; 70(10): e20240532, 2024.
Artigo em Inglês | MEDLINE | ID: mdl-39356958

RESUMO

OBJECTIVE: The aim of this study was to investigate the role of systemic immune-inflammation index, neutrophil-lymphocyte ratio, lymphocyte-monocyte ratio, and platelet-lymphocyte ratios calculated in the first trimester as inflammatory markers in predicting gestational diabetes mellitus diagnosis. METHODS: This study was conducted retrospectively at a tertiary center between January 2020 and June 2023. A total of 111 pregnant women with gestational diabetes and 378 pregnant women in the control group were included in the study. Systemic immune-inflammation index, neutrophil-lymphocyte ratio, lymphocyte-monocyte ratio, and platelet-lymphocyte ratios values were compared between the gestational diabetes mellitus group patients and the healthy group. Receiver operating characteristic analysis curve was used for predicting gestational diabetes mellitus using systemic immune-inflammation index and lymphocyte-monocyte ratio. RESULTS: In pregnant women in the first trimester, systemic immune-inflammation index and lymphocyte-monocyte ratio values based on routine complete blood count parameters were found to be statistically significantly higher in gestational diabetes mellitus patients compared to healthy patients, while neutrophil-lymphocyte ratio and platelet-lymphocyte ratios values were found to be similar (p=0.033, p=0.005, p=0.211, and p=0.989). For predicting gestational diabetes mellitus, a cut-off value of 655.75 for systemic immune-inflammation index resulted in 80.2% sensitivity and 34.4% specificity, and a cut-off value of 3.62 for lymphocyte-monocyte ratio resulted in 56.8% sensitivity and 63.2% specificity, indicating good discriminatory ability. CONCLUSION: We believe that systemic immune-inflammation index and lymphocyte-monocyte ratio values measured in the first-trimester complete blood count parameters are effective in predicting gestational diabetes mellitus but are not effective in determining insulin requirement.


Assuntos
Biomarcadores , Diabetes Gestacional , Neutrófilos , Primeiro Trimestre da Gravidez , Curva ROC , Humanos , Diabetes Gestacional/sangue , Diabetes Gestacional/imunologia , Diabetes Gestacional/diagnóstico , Feminino , Gravidez , Primeiro Trimestre da Gravidez/sangue , Primeiro Trimestre da Gravidez/imunologia , Estudos Retrospectivos , Adulto , Biomarcadores/sangue , Inflamação/sangue , Inflamação/imunologia , Linfócitos/imunologia , Valor Preditivo dos Testes , Monócitos/imunologia , Estudos de Casos e Controles , Contagem de Plaquetas , Contagem de Linfócitos , Sensibilidade e Especificidade
3.
Trop Anim Health Prod ; 56(8): 314, 2024 Oct 02.
Artigo em Inglês | MEDLINE | ID: mdl-39356340

RESUMO

Bovine viral diarrhea virus (BVDV) causes ongoing economic losses to cattle industries, directly through reduced herd performance or indirectly through control program costs. ELISA assays, one of the most widely used techniques due to their ease of implementation, have been a valuable tool for mass surveillance and detection of BVDV. In this study, we developed a new indirect ELISA (rE2-ELISA) for serologic detection of BVDV. The assay considers three recombinant E2 protein subtypes as antigens, allowing serologic diagnosis of BVDV-1b (high prevalence worldwide), BVDV-1d and 1e (high prevalence in southern Chile) sub-genotypes. Recombinant E2 (rE2) proteins were successfully expressed in stably transfected CHO cells. Conditions for rE2 ELISAs were established after determining appropriate concentrations of antigen, blocking agent, secondary antibody, and serum dilutions to achieve maximum discrimination between positive and negative serum samples. The developed rE2-ELISA showed a sensitivity of 92.86% and a specificity of 98.33%. Clinical testing of 180 serum samples from herds in southern Chile showed high accuracy (kappa > 0.8) compared to the commercial BVDV Total Ab kit (IDEXX), with 95.37% positive and 87.5% negative predictive value. In addition, the rE2 ELISA has shown the capability to detect anti-BVDV antibodies from naturally infected animals with sub-genotypes 1b, 1e, or undetermined. These results indicate that the developed indirect ELISA could serve as a valid, and efficient alternative for identifying BVDV-infected animals, thus contributing to the success of disease control and eradication programs.


Assuntos
Doença das Mucosas por Vírus da Diarreia Viral Bovina , Ensaio de Imunoadsorção Enzimática , Sensibilidade e Especificidade , Animais , Ensaio de Imunoadsorção Enzimática/veterinária , Ensaio de Imunoadsorção Enzimática/métodos , Bovinos , Doença das Mucosas por Vírus da Diarreia Viral Bovina/diagnóstico , Doença das Mucosas por Vírus da Diarreia Viral Bovina/sangue , Doença das Mucosas por Vírus da Diarreia Viral Bovina/virologia , Chile , Genótipo , Vírus da Diarreia Viral Bovina Tipo 1/imunologia , Vírus da Diarreia Viral Bovina Tipo 1/isolamento & purificação , Vírus da Diarreia Viral Bovina/imunologia , Vírus da Diarreia Viral Bovina/isolamento & purificação , Proteínas do Envelope Viral/imunologia , Proteínas do Envelope Viral/genética , Antígenos Virais/imunologia , Cricetulus , Células CHO , Anticorpos Antivirais/sangue , Proteínas Recombinantes/imunologia
4.
Artigo em Inglês | MEDLINE | ID: mdl-39380589

RESUMO

Objective: To conduct a systematic review of external validation studies on the use of different Artificial Intelligence algorithms in breast cancer screening with mammography. Data source: Our systematic review was conducted and reported following the PRISMA statement, using the PubMed, EMBASE, and Cochrane databases with the search terms "Artificial Intelligence," "Mammography," and their respective MeSH terms. We filtered publications from the past ten years (2014 - 2024) and in English. Study selection: A total of 1,878 articles were found in the databases used in the research. After removing duplicates (373) and excluding those that did not address our PICO question (1,475), 30 studies were included in this work. Data collection: The data from the studies were collected independently by five authors, and it was subsequently synthesized based on sample data, location, year, and their main results in terms of AUC, sensitivity, and specificity. Data synthesis: It was demonstrated that the Area Under the ROC Curve (AUC) and sensitivity were similar to those of radiologists when using independent Artificial Intelligence. When used in conjunction with radiologists, statistically higher accuracy in mammogram evaluation was reported compared to the assessment by radiologists alone. Conclusion: AI algorithms have emerged as a means to complement and enhance the performance and accuracy of radiologists. They also assist less experienced professionals in detecting possible lesions. Furthermore, this tool can be used to complement and improve the analyses conducted by medical professionals.


Assuntos
Inteligência Artificial , Neoplasias da Mama , Mamografia , Mamografia/métodos , Humanos , Feminino , Neoplasias da Mama/diagnóstico por imagem , Detecção Precoce de Câncer/métodos , Sensibilidade e Especificidade , Algoritmos , Estudos de Validação como Assunto
5.
Rev Assoc Med Bras (1992) ; 70(10): e20240660, 2024.
Artigo em Inglês | MEDLINE | ID: mdl-39383393

RESUMO

OBJECTIVE: The objective of this study was to investigate serum Metrnl levels in pregnant women with gestational diabetes mellitus and compare them with pregnant women without gestational diabetes mellitus. METHODS: The gestational diabetes mellitus group consisted of 87 pregnant women diagnosed with gestational diabetes mellitus, and the control group consisted of 93 healthy pregnant women without gestational diabetes mellitus. Serum Metrnl levels were determined by the enzyme-linked immunosorbent assay method. RESULTS: The two groups were similar in terms of demographic features. The median serum Metrnl level was found to be 1.16 ng/mL in the gestational diabetes mellitus group, while it was determined as 2.2 ng/mL in the control group (p=0.001). The two groups were divided into two subgroups based on participants' body mass index, normal weight and overweight. The lowest median Metrnl level was detected in the normal weight gestational diabetes mellitus group, followed by the overweight gestational diabetes mellitus group, normal weight control group, and overweight control group (1.1, 1.2, 2, and 2.4 ng/mL, respectively). Receiver operating curve analysis was performed to determine the value of the serum Metrnl level in terms of predicting gestational diabetes mellitus. The area under the curve analysis of serum Metrnl for gestational diabetes mellitus estimation was 0.768 (p=0.000, 95%CI 0.698-0.839). The optimal cutoff value for serum Metrnl level was determined as 1.53 ng/mL with 69% sensitivity and 70% specificity. CONCLUSION: Serum Metrnl levels in pregnant women with gestational diabetes mellitus were found to be significantly lower than in pregnant women without gestational diabetes mellitus. The mechanisms underlying the decrease in serum Metrnl levels in gestational diabetes mellitus remain unclear for now, and future studies will reveal the role of Metrnl in the pathophysiology of gestational diabetes mellitus.


Assuntos
Biomarcadores , Índice de Massa Corporal , Diabetes Gestacional , Ensaio de Imunoadsorção Enzimática , Humanos , Diabetes Gestacional/sangue , Feminino , Gravidez , Adulto , Estudos Prospectivos , Estudos de Casos e Controles , Biomarcadores/sangue , Curva ROC , Valores de Referência , Adulto Jovem , Sobrepeso/sangue , Sensibilidade e Especificidade , Adipocinas
6.
Mem Inst Oswaldo Cruz ; 119: e230236, 2024.
Artigo em Inglês | MEDLINE | ID: mdl-39383402

RESUMO

BACKGROUND: During the coronavirus disease 19 (COVID-19) pandemic, diagnostic testing of the general population proved challenging due to limitations of the gold-standard diagnostic procedure using reverse transcription real-time polymerase chain reaction (RT-qPCR) for large-scale testing on the centralised model, especially in low-resource areas. OBJECTIVES: To address this, a point-of-care (PoC) diagnostic protocol for COVID-19 was developed, providing fast, reliable, and affordable testing, particularly for low-mid develop areas. METHODS: The PoC diagnostic process combines a simple paper-based RNA extraction method housed within a 3D-printed plastic device with a colorimetric reverse transcription loop-mediated isothermal amplification (RT-LAMP) assay. Nasopharyngeal/oropharyngeal swabs (NOS) and saliva samples were tested between 2020 and 2021, with the assistance of Santa Catarina's State Health Secretary, Brazil. FINDINGS: The developed diagnostic protocol showed a limit of detection of 9,900 copies and an overall diagnostic specificity of 98% for severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) from 1,348 clinical analysed samples. The diagnostic sensitivity was 95% for NOS samples, 85% for early morning saliva, and 69% for indiscriminate saliva. MAIN CONCLUSIONS: In conclusion, the developed device successfully extracted SARS-CoV-2 viral RNA from swabs and saliva clinical samples. When combined with colorimetric RT-LAMP, it provides results within 45 min using minimal resources, thus delivering a diagnostic kit protocol that is applicable in large-scale sampling.


Assuntos
COVID-19 , Técnicas de Diagnóstico Molecular , Técnicas de Amplificação de Ácido Nucleico , Testes Imediatos , SARS-CoV-2 , Saliva , Sensibilidade e Especificidade , Humanos , COVID-19/diagnóstico , SARS-CoV-2/genética , SARS-CoV-2/isolamento & purificação , Técnicas de Diagnóstico Molecular/métodos , Técnicas de Amplificação de Ácido Nucleico/métodos , Saliva/virologia , RNA Viral/análise , RNA Viral/isolamento & purificação , Teste de Ácido Nucleico para COVID-19/métodos , Pandemias , Brasil , Nasofaringe/virologia , Reprodutibilidade dos Testes , Teste para COVID-19/métodos
7.
BMC Pregnancy Childbirth ; 24(1): 628, 2024 Oct 01.
Artigo em Inglês | MEDLINE | ID: mdl-39354367

RESUMO

OBJECTIVE: This study introduces the complete blood count (CBC), a standard prenatal screening test, as a biomarker for diagnosing preeclampsia with severe features (sPE), employing machine learning models. METHODS: We used a boosting machine learning model fed with synthetic data generated through a new methodology called DAS (Data Augmentation and Smoothing). Using data from a Brazilian study including 132 pregnant women, we generated 3,552 synthetic samples for model training. To improve interpretability, we also provided a ridge regression model. RESULTS: Our boosting model obtained an AUROC of 0.90±0.10, sensitivity of 0.95, and specificity of 0.79 to differentiate sPE and non-PE pregnant women, using CBC parameters of neutrophils count, mean corpuscular hemoglobin (MCH), and the aggregate index of systemic inflammation (AISI). In addition, we provided a ridge regression equation using the same three CBC parameters, which is fully interpretable and achieved an AUROC of 0.79±0.10 to differentiate the both groups. Moreover, we also showed that a monocyte count lower than 490 / m m 3 yielded a sensitivity of 0.71 and specificity of 0.72. CONCLUSION: Our study showed that ML-powered CBC could be used as a biomarker for sPE diagnosis support. In addition, we showed that a low monocyte count alone could be an indicator of sPE. SIGNIFICANCE: Although preeclampsia has been extensively studied, no laboratory biomarker with favorable cost-effectiveness has been proposed. Using artificial intelligence, we proposed to use the CBC, a low-cost, fast, and well-spread blood test, as a biomarker for sPE.


Assuntos
Biomarcadores , Aprendizado de Máquina , Pré-Eclâmpsia , Humanos , Pré-Eclâmpsia/diagnóstico , Pré-Eclâmpsia/sangue , Feminino , Gravidez , Biomarcadores/sangue , Contagem de Células Sanguíneas/métodos , Adulto , Sensibilidade e Especificidade , Brasil , Índice de Gravidade de Doença , Curva ROC , Diagnóstico Pré-Natal/métodos
8.
Biosensors (Basel) ; 14(9)2024 Aug 27.
Artigo em Inglês | MEDLINE | ID: mdl-39329791

RESUMO

The COVID-19 pandemic highlighted testing inequities in developing countries. Lack of lateral flow test (LFT) manufacturing capacity was a major COVID-19 response bottleneck in low- and middle-income regions. Here we report the development of an open-access LFT for SARS-CoV-2 detection comparable to commercial tests that requires only locally available supplies. The main critical resource is a locally developed horse polyclonal antibody (pAb) whose sensitivity and selectivity are greatly enhanced by affinity purification. We demonstrate that these Abs can perform similarly to commercial monoclonal antibodies (mAbs), as well as mAbs and other pAbs developed against the same antigen. We report a workflow for test optimization using nasopharyngeal swabs collected for RT-qPCR, spiked with the inactivated virus to determine analytical performance characteristics as the limit of detection, among others. Our final prototype showed a performance similar to available tests (sensitivity of 83.3% compared to RT-qPCR, and 90.9% compared to commercial antigen tests). Finally, we discuss the possibility and the challenges of utilizing affinity-purified pAbs as an alternative for the local development of antigen tests in an outbreak context and as a tool to address inequalities in access to rapid tests.


Assuntos
COVID-19 , SARS-CoV-2 , SARS-CoV-2/isolamento & purificação , Humanos , COVID-19/diagnóstico , COVID-19/virologia , Anticorpos Monoclonais , Anticorpos Antivirais , Sensibilidade e Especificidade , Animais
9.
Mycoses ; 67(9): e13793, 2024 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-39239746

RESUMO

Sporotrichosis diagnosis involves a series of analyses, including culture and antibody detection in serum samples. Serologic methods may sometimes yield false-negative or false-positive results, leading to inaccurate diagnoses. This study assessed specific patient groups in which antibody detection of different isotypes and subclasses may lack sensitivity. An enzyme-linked immunosorbent assay (ELISA) with Sporothrix brasiliensis exoantigens was used to investigate IgM, IgG, IgG1, IgG2, IgG3, IgG4, IgA, IgA1 and IgA2 antibodies in human serum samples. Eighty serum samples from patients with different sporotrichosis clinical manifestations, including cutaneous forms with and without hypersensitivity manifestations, extracutaneous forms (bone, ocular, meningeal and pulmonary), disseminated cutaneous forms and disseminated forms in individuals living with HIV/AIDS, diabetics and alcoholics, were evaluated. The ELISA sensitivities in the detection of different antibodies ranged from 0.85 to 0.60 for the detection of IgG2 and IgG3, respectively. The antibodies with higher area under ROC curves were IgG2, IgG, IgA and IgA1. There were no significant differences in the immunological reactivity of the tested antibodies among different clinical forms of sporotrichosis. The data revealed a higher likelihood of a false-negative outcome in patients with lesions in the nasal mucosa regarding the detection of IgM and a lower likelihood in patients with lymphocutaneous sporotrichosis regarding the detection of IgG3. Patients with hypersensitivity manifestations had a 3.71 odds ratio to yield negative results in total IgG detection. In conclusion, we identified specific patient groups in which antibody detection may lack sensitivity, thus contributing to a better understanding of the diagnostic challenges associated with this condition.


Assuntos
Anticorpos Antifúngicos , Ensaio de Imunoadsorção Enzimática , Sensibilidade e Especificidade , Sporothrix , Esporotricose , Humanos , Esporotricose/imunologia , Esporotricose/diagnóstico , Anticorpos Antifúngicos/sangue , Sporothrix/imunologia , Sporothrix/classificação , Masculino , Feminino , Adulto , Pessoa de Meia-Idade , Isotipos de Imunoglobulinas/sangue , Isotipos de Imunoglobulinas/imunologia , Imunoglobulina G/sangue , Idoso , Adulto Jovem , Antígenos de Fungos/imunologia , Antígenos de Fungos/sangue , Imunoglobulina A/sangue , Imunoglobulina M/sangue
10.
Rev Assoc Med Bras (1992) ; 70(9): e20240523, 2024.
Artigo em Inglês | MEDLINE | ID: mdl-39230068

RESUMO

OBJECTIVE: The primary objective was to assess the diagnostic accuracy of a deep learning-based artificial intelligence model for the detection of acute appendicular fractures in pediatric patients presenting with a recent history of trauma to the emergency department. The secondary goal was to examine the effect of assistive support on the emergency doctor's ability to detect fractures. METHODS: The dataset was 5,150 radiographs of which 850 showed fractures, while 4,300 radiographs did not show any fractures. The process utilized 4,532 (88%) radiographs, inclusive of both fractured and non-fractured radiographs, in the training phase. Subsequently, 412 (8%) radiographs were appraised during validation, and 206 (4%) were set apart for the testing phase. With and without artificial intelligence assistance, the emergency doctor reviewed another set of 2,000 radiographs (400 fractures and 600 non-fractures each) for labeling in the second test. RESULTS: The artificial intelligence model showed a mean average precision 50 of 89%, a specificity of 92%, a sensitivity of 90%, and an F1 score of 90%. The confusion matrix revealed that the model trained with artificial intelligence achieved accuracies of 93 and 95% in detecting fractures, respectively. Artificial intelligence assistance improved the reading sensitivity from 93.7% (without assistance) to 97.0% (with assistance) and the reading accuracy from 88% (without assistance) to 94.9% (with assistance). CONCLUSION: A deep learning-based artificial intelligence model has proven to be highly effective in detecting fractures in pediatric patients, enhancing the diagnostic capabilities of emergency doctors through assistive support.


Assuntos
Inteligência Artificial , Fraturas Ósseas , Humanos , Fraturas Ósseas/diagnóstico por imagem , Criança , Pré-Escolar , Sensibilidade e Especificidade , Feminino , Aprendizado Profundo , Serviço Hospitalar de Emergência , Masculino , Reprodutibilidade dos Testes , Radiografia/métodos , Adolescente , Lactente
11.
Rev Assoc Med Bras (1992) ; 70(8): e20240370, 2024.
Artigo em Inglês | MEDLINE | ID: mdl-39230144

RESUMO

OBJECTIVE: In the hepatitis C virus (HCV) diagnostic algorithm, an anti-HCV screening test is recommended first. In countries with low HCV prevalence, anti-HCV testing can often give false-positive results. This may lead to unnecessary retesting, increased costs, and psychological stress for patients. METHODS: In this study, the most appropriate S/Co (signal-cutoff) value to predict HCV viremia in anti-HCV test(+) individuals was determined, and the effect of genotype differences was evaluated. Of the 96,515 anti-HCV tests performed between 2020 and 2023, 934 were reactive. A total of 332 retests and 65 patients without HCV-ribonucleic acid (RNA) analysis were excluded. Demographic data were calculated for 537 patients, and 130 patients were included in the study. RESULTS: The average age of 537 patients was 55±18 years, and 57.1% were women. The anti-HCV positivity rate was 0.62% (602/96,515), and the actual anti-HCV positivity rate was 0.13% (130/96,515). Anti-HCV levels were higher in HCV-RNA(+) patients than in HCV-RNA-negative individuals (p<0.0001) (Table 1). Receiver operating characteristic curve analysis identified the optimal S/Co value to be 10.86 to identify true positive cases. Sensitivity was 96.1%, specificity was 61.2%, positive predictive value (PPV) was 44.2%, and negative predictive value (NPV) was 98% (Figure 2). A total of 107 (82.3%) of the patients were identified as GT1, and the most common subtype was GT1b (n=100). CONCLUSION: If anti-HCV S/Co is ≥10.86, direct HCV RNA testing may be recommended; However, the possibility of false positivity should be considered in patients with a S/Co value below 10.86.


Assuntos
Genótipo , Hepacivirus , Anticorpos Anti-Hepatite C , Hepatite C , Valor Preditivo dos Testes , RNA Viral , Viremia , Humanos , Feminino , Masculino , Pessoa de Meia-Idade , Hepacivirus/genética , RNA Viral/sangue , RNA Viral/análise , Anticorpos Anti-Hepatite C/sangue , Hepatite C/genética , Hepatite C/sangue , Adulto , Idoso , Sensibilidade e Especificidade
12.
Mem Inst Oswaldo Cruz ; 119: e230183, 2024.
Artigo em Inglês | MEDLINE | ID: mdl-39292107

RESUMO

BACKGROUND: PPE 59, which is absent from bacillus Calmette Guérin (BCG) strains, seems to induce a humoral immune response in patients with tuberculosis (TB). Additional studies are needed to better evaluate this protein in immune response to tuberculosis. OBJECTIVES: To evaluate the response of antibodies to PPE59 in TB individuals, its combination with IgG response to other, previously tested mycobacterial antigens (Ag) and with sputum smear microbiology (SM) results. METHODS: We have cloned and expressed the rv3429 gene that encodes PPE59, then IgG, IgM, and IgA against PPE59 antigens measured by enzyme-linked immunosorbent assay (ELISA) in 212 sera samples obtained from the following subject cohorts: TB residents from Italy (79) and in Brazil (52); and an all-Brazilian cohort of 55 patients with other respiratory disorders; 10 patients infected with non-tuberculous mycobacteria, and 16 asymptomatic subjects. Drawing on results from a previous study(17) of serum samples from Brazilian subjects tested for IgG by ELISA against mycobacterial antigens ESAT-6, 16kDa, MT10.3, MPT-64 and 38kDa, the results were analysed in combination with those of the PPE59 and SM tests. FINDINGS: Keeping the specificity rate at 97%, the overall PPE59 IgA sensitivity was 42.7%, while IgG and IgM showed lower performance (p < 0.0001). Combining PPE59 IgA/16kDa IgG results increased sensitivity to 71%, and even higher rates when the results were combined with SM results (86.5%, p = 0.001), at 88.9% specificity. Positive IgA was associated with pulmonary image alterations of high TB probability (p < 0.05). MAIN CONCLUSIONS: Tests with TB patients found a moderate frequency of positivity for PPE59 IgA. However, the higher level of sensitivity attained in combination with PPE59 IgA/16kDa IgG/SM results unheard of before, although imperfect, suggests that this may be a potential additional tool for rapid detection of TB in low-resource areas.


Assuntos
Anticorpos Antibacterianos , Antígenos de Bactérias , Biomarcadores , Ensaio de Imunoadsorção Enzimática , Imunoglobulina A , Imunoglobulina G , Imunoglobulina M , Humanos , Anticorpos Antibacterianos/sangue , Antígenos de Bactérias/imunologia , Imunoglobulina G/sangue , Imunoglobulina M/sangue , Masculino , Feminino , Biomarcadores/sangue , Adulto , Imunoglobulina A/sangue , Sensibilidade e Especificidade , Pessoa de Meia-Idade , Mycobacterium tuberculosis/imunologia , Tuberculose/diagnóstico , Tuberculose/imunologia , Tuberculose/sangue , Adulto Jovem , Tuberculose Pulmonar/diagnóstico , Tuberculose Pulmonar/imunologia , Tuberculose Pulmonar/sangue , Escarro/microbiologia , Brasil , Proteínas de Bactérias/imunologia , Idoso , Adolescente , Estudos de Coortes
13.
Rev Assoc Med Bras (1992) ; 70(9): e20240696, 2024.
Artigo em Inglês | MEDLINE | ID: mdl-39292092

RESUMO

OBJECTIVE: In endometrial cancer surgery, sentinel lymph node dissection is used instead of staging surgery, particularly in advanced disease that is limited to the uterus. The aim of this study is to evaluate our practice of robotic sentinel lymph node dissection, which is applied to endometrial cancer patients in our tertiary cancer treatment center, according to the current literature, and to share our own data. METHODS: Included in our analysis are patients who underwent robotic sentinel lymph node dissection for endometrial cancer utilizing indocyanine green in our center between January 2018 and January 2024. RESULTS: In all, of the 93 endometrial carcinoma patients who underwent sentinel lymph node biopsy, 63 were classified as low-risk, while 30 were high-risk according to the European Society of Gynaecological Oncology and National Comprehensive Cancer Network guidelines. We found sentinel lymph nodes in both low-risk and high-risk patients, with an overall sensitivity of 96.32% (95% confidence interval [CI], 85.12-99.71), specificity of 100% (95%CI, 92.20-99.8), negative predictive value of 96.72% (95%CI, 87.03-99.89), and negative likelihood ratio of 0.06 (95%CI, 0.01-0.36). CONCLUSION: After evaluating our data retrospectively, we determined that we were compatible with the current literature.


Assuntos
Neoplasias do Endométrio , Procedimentos Cirúrgicos Robóticos , Biópsia de Linfonodo Sentinela , Centros de Atenção Terciária , Humanos , Feminino , Neoplasias do Endométrio/cirurgia , Neoplasias do Endométrio/patologia , Procedimentos Cirúrgicos Robóticos/métodos , Pessoa de Meia-Idade , Biópsia de Linfonodo Sentinela/métodos , Idoso , Estudos Retrospectivos , Adulto , Sensibilidade e Especificidade , Linfonodo Sentinela/patologia , Linfonodo Sentinela/cirurgia , Excisão de Linfonodo/métodos , Verde de Indocianina , Estadiamento de Neoplasias , Idoso de 80 Anos ou mais , Metástase Linfática
14.
Int J Mol Sci ; 25(18)2024 Sep 11.
Artigo em Inglês | MEDLINE | ID: mdl-39337297

RESUMO

We developed a protein to rapidly and accurately diagnose Chagas disease, a life-threatening illness identified by the WHO as a critical worldwide public health risk. Limitations in present day serological tests are complicating the current health situation and contributing to most infected persons being unaware of their condition and therefore untreated. To improve diagnostic testing, we developed an immunological mimic of the etiological agent, Trypanosoma cruzi, by combining ten pathogen-specific epitopes within the beta-barrel protein structure of Thermal Green Protein. The resulting multi-epitope protein, DxCruziV3, displayed high specificity and sensitivity as the antibody capture reagent in an ELISA platform with an analytical sensitivity that exceeds WHO recommendations. Within an immunochromatographic platform, DxCruziV3 showed excellent performance for the point of application diagnosis in a region endemic for multiple diseases, the municipality of Barcelos in the state of Amazonas, Brazil. In total, 167 individuals were rapidly tested using whole blood from a finger stick. As recommended by the Brazilian Ministry of Health, venous blood samples were laboratory tested by conventional assays for comparison. Test results suggest utilizing DxCruziV3 in different assay platforms can confidently diagnose chronic infections by T. cruzi. Rapid and more accurate results will benefit everyone but will have the most noticeable impact in resource-limited rural areas where the disease is endemic.


Assuntos
Doença de Chagas , Ensaio de Imunoadsorção Enzimática , Epitopos , Testes Sorológicos , Trypanosoma cruzi , Doença de Chagas/diagnóstico , Doença de Chagas/sangue , Doença de Chagas/imunologia , Humanos , Ensaio de Imunoadsorção Enzimática/métodos , Trypanosoma cruzi/imunologia , Testes Sorológicos/métodos , Epitopos/imunologia , Doença Crônica , Masculino , Sensibilidade e Especificidade , Feminino , Adulto , Anticorpos Antiprotozoários/sangue , Anticorpos Antiprotozoários/imunologia , Pessoa de Meia-Idade , Antígenos de Protozoários/imunologia , Antígenos de Protozoários/sangue , Brasil/epidemiologia
15.
Acta Trop ; 259: 107382, 2024 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-39244140

RESUMO

The diagnosis of Chagas disease mostly relies on the use of multiple serologic tests that are often unavailable in many of the remote settings where the disease is highly prevalent. In the Teniente Irala Fernández Municipality, in central Paraguay, efforts have been made to increase the diagnostic capabilities of specific rural health centres, but no quality assurance of the results produced has been performed. We comparatively analysed the results obtained with 300 samples tested using a commercial rapid diagnostic test (RDT) and enzyme linked immunosorbent assays (ELISA) at the laboratory of the Teniente Irala Fernández Health Center (CSTIF) with those generated upon repeating the tests at an independent well-equipped research laboratory (CEDIC). A subgroup of 52 samples were further tested at Paraguay's Central Public Health Laboratory (LCSP) by means of a different technique to evaluate the diagnostic performance of the tests carried out at CSTIF. We observed an excellent agreement between the ELISA results obtained at CSTIF and CEDIC (kappa coefficients between 0.85 and 0.93 for every kit evaluated), and an overall good performance of the tests carried out at CSTIF. However, the sensitivity of one kit was lower at CSTIF (81.3 %) than at CEDIC (100 %). The individual use of an RDT to detect the infection at CSTIF showed a similar sensitivity to that obtained combining it to an ELISA test (92.3% vs 88.5, p = 1). Nonetheless, the generalizability of this result is yet limited and will require of further studies.


Assuntos
Doença de Chagas , Atenção Primária à Saúde , População Rural , Sensibilidade e Especificidade , Testes Sorológicos , Paraguai , Humanos , Doença de Chagas/diagnóstico , Testes Sorológicos/métodos , Testes Sorológicos/normas , Adulto , Masculino , Feminino , Garantia da Qualidade dos Cuidados de Saúde , Adolescente , Ensaio de Imunoadsorção Enzimática/métodos , Ensaio de Imunoadsorção Enzimática/normas , Pessoa de Meia-Idade , Criança , Adulto Jovem , Pré-Escolar , Idoso , Anticorpos Antiprotozoários/sangue
16.
Sci Rep ; 14(1): 22520, 2024 09 28.
Artigo em Inglês | MEDLINE | ID: mdl-39342022

RESUMO

Monitoring yellow fever in non-human primates (NHPs) is an early warning system for sylvatic yellow fever outbreaks, aiding in preventing human cases. However, current diagnostic tests for this disease, primarily relying on RT-qPCR, are complex and costly. Therefore, there is a critical need for simpler and more cost-effective methods to detect yellow fever virus (YFV) infection in NHPs, enabling early identification of viral circulation. In this study, an RT-LAMP assay for detecting YFV in NHP samples was developed and validated. Two sets of RT-LAMP primers targeting the YFV NS5 and E genes were designed and tested together with a third primer set to the NS1 locus using NHP tissue samples from Southern Brazil. The results were visualized by colorimetry and compared to the RT-qPCR test. Standardization and validation of the RT-LAMP assay demonstrated 100% sensitivity and specificity compared to RT-qPCR, with a detection limit of 12 PFU/mL. Additionally, the cross-reactivity test with other flaviviruses confirmed a specificity of 100%. Our newly developed RT-LAMP diagnostic test for YFV in NHP samples will significantly contribute to yellow fever monitoring efforts, providing a simpler and more accessible method for viral early detection. This advancement holds promise for enhancing surveillance and ultimately preventing the spread of yellow fever.


Assuntos
Técnicas de Amplificação de Ácido Nucleico , Sensibilidade e Especificidade , Febre Amarela , Vírus da Febre Amarela , Animais , Vírus da Febre Amarela/genética , Vírus da Febre Amarela/isolamento & purificação , Brasil/epidemiologia , Febre Amarela/diagnóstico , Febre Amarela/virologia , Febre Amarela/epidemiologia , Técnicas de Amplificação de Ácido Nucleico/métodos , Técnicas de Diagnóstico Molecular/métodos , Primatas/virologia
17.
Parasitol Res ; 123(9): 312, 2024 Sep 02.
Artigo em Inglês | MEDLINE | ID: mdl-39218957

RESUMO

Abdominal angiostrongyliasis (AA) is a zoonotic and severe parasitic infection caused by Angiostrongylus costaricensis. AA is currently diagnosed by the observation of A. costaricensis-compatible structures in biopsies or the detection of antibodies in serological tests. However, molecular methods targeting homologous sequences of A. costaricensis have not been designed before, and therefore, an HRM-coupled qPCR was developed to detect the internal transcribed spacer 1 (ITS1) of the parasite. The present assay successfully amplified DNA of A. costaricensis obtained from different hosts and identified slight sequence differences through the HRM analysis. The detection limit of the HRM-qPCR was 0.00036 ng/µL, 1.0 ng/µL, and 0.1 ng/µL when A. costaricensis DNA was diluted in nuclease-free water, whole blood, and sera, respectively, which highlights its potential use for cell-free DNA detection. Moreover, the reaction did not cross-amplify DNA of Angiostrongylus cantonensis, Strongyloides stercoralis, and other nematodes, thus emphasizing its specificity. Additionally, the assay tested positive in formalin-fixed paraffin embedded biopsies with visible A. costaricensis adults or eggs, but not in samples without evident parasites or a low number of larvae, which suggests that the reaction is useful for confirming the presence of the nematode in clinical samples. Finally, DNA of sera from patients with AA was evaluated with the HRM-qPCR but none tested positive, possibly due to long storage periods of the samples which could have led to cfDNA degradation. These results indicate that this assay may be useful in the confirmation of AA and its prospection for cell-free DNA detection protocols.


Assuntos
Angiostrongylus , DNA de Helmintos , DNA Espaçador Ribossômico , Reação em Cadeia da Polimerase em Tempo Real , Sensibilidade e Especificidade , Infecções por Strongylida , Animais , Reação em Cadeia da Polimerase em Tempo Real/métodos , Angiostrongylus/genética , Angiostrongylus/isolamento & purificação , Infecções por Strongylida/diagnóstico , Infecções por Strongylida/parasitologia , DNA Espaçador Ribossômico/genética , DNA de Helmintos/genética , Humanos , Temperatura de Transição , Técnicas de Diagnóstico Molecular/métodos
18.
Vet Res Commun ; 48(6): 4185-4189, 2024 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-39340735

RESUMO

Bovine Genital Leptospirosis (BGL) is a chronic reproductive syndrome characterized by genital infection by Leptospira spp. An accurate diagnosis of BGL is crucial to implementing proper control measures in field conditions. This study aimed to evaluate the reliability of serology by Microscopic Agglutination Test (MAT) for diagnosing leptospirosis in subfertile cows with genital infection. Of three herds, 93 non-pregnant cows with reproductive failures were submitted to the blood sampling (serology by MAT) and genital samples (lipL32-PCR). A total of 62/93 (66.6%) cows presented seroreactive to cutoff 100, while 45/93 (48.4%) cows were positive to cutoff 200, mainly against the Sejroe serogroup. In PCR analysis, 55/93 (59.1%) were positive. MAT results were compared with PCR (considered the standard), and test parameters and Cohen's kappa (ƙ) were calculated for the cut-offs 100 and 200. A ROC curve was performed for each cut-off of titers 100 to 1,600. The sensitivity and specificity of MAT100 were calculated at 66.6% and 33.3%, while for MAT200 the sensitivity was estimated as 35% and specificity as 54.5%. The accuracy of MAT was poor, being 54.8% in MAT100 and 42% in MAT200. Furthermore, the area under the curve of ROC analysis was low for all titers, and the correlation was poor for MAT100 and MAT200 (ƙ < 0). The results demonstrated that MAT is a limited technique to diagnose bovine genital carriers individually, and if only MAT is applied, genital carriers may pass undetected, impairing the control programs.


Assuntos
Testes de Aglutinação , Doenças dos Bovinos , Leptospirose , Sensibilidade e Especificidade , Animais , Leptospirose/veterinária , Leptospirose/diagnóstico , Bovinos , Testes de Aglutinação/veterinária , Feminino , Doenças dos Bovinos/diagnóstico , Doenças dos Bovinos/microbiologia , Doenças dos Genitais Femininos/veterinária , Doenças dos Genitais Femininos/diagnóstico , Doenças dos Genitais Femininos/microbiologia , Leptospira/isolamento & purificação , Reprodutibilidade dos Testes
19.
Clin Oral Investig ; 28(10): 531, 2024 Sep 19.
Artigo em Inglês | MEDLINE | ID: mdl-39298025

RESUMO

AIM: To evaluate the influence of different levels of metal artifact reduction (MAR) tool and milliamperage (mA) on the diagnosis of fracture extension in endodontically treated teeth using cone beam CT (CBCT). MATERIALS AND METHODS: Ten maxillary premolars were endodontically treated and positioned in the empty sockets of a human maxilla covered with wax. CBCT acquisitions were performed using the Eagle Edge device (Dabi Atlante, Brazil) adjusted to 120 kVp, FOV of 4 × 6 cm, exposure time of 24 s and voxel size of 0.2 mm in 8 different conditions with different MAR (1, 2 and 3) and mA (3.2 and 6.3) levels. Crown-root fractures were simulated in the universal testing machine, and CBCT images were acquired again. Five radiologists evaluated the presence and extension of fractures with a 5-point scale. Statistical analysis was performed by analysis of variance, Tukey and Kappa test (α = 0.05). RESULTS: Although different mA levels did not significantly (p > 0.05) affect the diagnosis values for fracture presence and extension, when evaluated the different levels of MAR, AUC and sensitivity showed significantly higher values (p < 0.05) for MAR 0 using 6.3 mA and kappa agreement showed significantly higher values (p < 0.05) for MAR 0 and 2 using 6.3 mA. CONCLUSIONS: Although mA levels do not have a diagnostic effect when isolating the MAR level; in 6.3 mA, MAR 0 and 2 can positively influence the diagnosis of fracture extension in endodontically treated teeth using CBCT. CLINICAL RELEVANCE: The isolate evaluation of dental fracture presence can overlook diagnostics error of its extension.


Assuntos
Artefatos , Dente Pré-Molar , Tomografia Computadorizada de Feixe Cônico , Fraturas dos Dentes , Dente não Vital , Humanos , Fraturas dos Dentes/diagnóstico por imagem , Tomografia Computadorizada de Feixe Cônico/métodos , Dente não Vital/diagnóstico por imagem , Dente Pré-Molar/diagnóstico por imagem , Dente Pré-Molar/lesões , Técnicas In Vitro , Metais , Maxila/diagnóstico por imagem , Sensibilidade e Especificidade
20.
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