RESUMEN
El priapismo es una erección dolorosa y persistente acompañada o no de estímulo sexual. Una causa poco frecuente de esta anormalidad es la leucemia mieloide crónica. Se han reportado pocos casos de priapismo como manifestación inicial de una leucemia de este tipo en pacientes adolescentes. A continuación, se informa el caso de un paciente de 16 años de edad que presentó priapismo como manifestación inicial de una leucemia mieloide crónica. Durante su evolución, no se realizó aspiración de los cuerpos cavernosos. Se inició tratamiento hematológico específico y, ante la persistencia del priapismo, fue necesario realizar un shunt de cuerpos cavernosos en dos ocasiones, tratamiento a pesar del cual existen altas probabilidades de secuelas.
Priapism is a painful and persistent erection, with or without sexual stimulation. A rare cause of such abnormality is chronic myeloid leukemia. Few cases of priapism as an initial manifestation of this type of leukemia have been reported in adolescent patients. Here we describe the case of a 16-year-old patient who presented with priapism as the initial manifestation of chronic myeloid leukemia. No cavernosal aspiration was performed. A specific hematological treatment was started and, given the persistence of priapism, the patient required 2 corpora cavernosa shunt procedures; despite this treatment, there is a high probability of sequelae.
Asunto(s)
Humanos , Masculino , Adolescente , Priapismo/complicaciones , Priapismo/etiología , Leucemia Mielógena Crónica BCR-ABL Positiva/complicaciones , Leucemia Mielógena Crónica BCR-ABL Positiva/diagnóstico , Enfermedad CrónicaRESUMEN
Chronic myeloid leukemia (CML) is a type of leukemia whose main genetic marker is the reciprocal translocation that leads to the production of the BCR::ABL1 oncoprotein. The expression of some genes may interfere with the progression and development of leukemias. MicroRNAs are small non-coding RNAs that have the potential to alter the expression of some genes and may be correlated with some types of leukemia and could be used as biomarkers in the diagnosis and prognosis of patients. Therefore, this project carried out an analysis of microRNA-type plasma biomarkers in patients with chronic myeloid leukemia at unique points, including follow-up analysis of patients from the Erasto Gaertner Hospital. 35 microRNAs were analyzed in different cohorts. Inside those groups, 70 samples were analyzed at unique points and 11 patients in a follow-up analysis. Statistically different results were found for microRNA-7-5p, which was found to be upregulated in patients with high expression of the BCR::ABL1 transcript when compared to healthy controls. This microRNA also had evidence of behavior related to BCR::ABL1 when analyzed in follow-up, but strong evidence was not found. In this way, this work obtained results that may lead to manifestations of a relationship between miR-7-5p and chronic myeloid leukemia, and evaluations of possible microRNAs that are not related to this pathology.
Asunto(s)
Leucemia Mielógena Crónica BCR-ABL Positiva , MicroARNs , Humanos , MicroARNs/genética , Proteínas de Fusión bcr-abl/genética , Proteínas de Fusión bcr-abl/metabolismo , Leucemia Mielógena Crónica BCR-ABL Positiva/diagnóstico , Leucemia Mielógena Crónica BCR-ABL Positiva/genética , Leucemia Mielógena Crónica BCR-ABL Positiva/patología , Translocación Genética , BiomarcadoresRESUMEN
Priapism is a painful and persistent erection, with or without sexual stimulation. A rare cause of such abnormality is chronic myeloid leukemia. Few cases of priapism as an initial manifestation of this type of leukemia have been reported in adolescent patients. Here we describe the case of a 16-year-old patient who presented with priapism as the initial manifestation of chronic myeloid leukemia. No cavernosal aspiration was performed. A specific hematological treatment was started and, given the persistence of priapism, the patient required 2 corpora cavernosa shunt procedures; despite this treatment, there is a high probability of sequelae.
El priapismo es una erección dolorosa y persistente acompañada o no de estímulo sexual. Una causa poco frecuente de esta anormalidad es la leucemia mieloide crónica. Se han reportado pocos casos de priapismo como manifestación inicial de una leucemia de este tipo en pacientes adolescentes. A continuación, se informa el caso de un paciente de 16 años de edad que presentó priapismo como manifestación inicial de una leucemia mieloide crónica. Durante su evolución, no se realizó aspiración de los cuerpos cavernosos. Se inició tratamiento hematológico específico y, ante la persistencia del priapismo, fue necesario realizar un shunt de cuerpos cavernosos en dos ocasiones, tratamiento a pesar del cual existen altas probabilidades de secuelas.
Asunto(s)
Leucemia Mielógena Crónica BCR-ABL Positiva , Priapismo , Masculino , Humanos , Adolescente , Priapismo/etiología , Priapismo/complicaciones , Leucemia Mielógena Crónica BCR-ABL Positiva/complicaciones , Leucemia Mielógena Crónica BCR-ABL Positiva/diagnóstico , Enfermedad CrónicaRESUMEN
Chronic myeloid leukemia (CML) is a well-characterized oncological disease in which virtually all patients possess a translocation (9;22) that generates the tyrosine kinase BCR::ABL1 protein. This translocation represents one of the milestones in molecular oncology in terms of both diagnostic and prognostic evaluations. The molecular detection of the BCR::ABL1 transcription is a required factor for CML diagnosis, and its molecular quantification is essential for assessing treatment options and clinical approaches. In the CML molecular context, point mutations on the ABL1 gene are also a challenge for clinical guidelines because several mutations are responsible for tyrosine kinase inhibitor resistance, indicating that a change may be necessary in the treatment protocol. So far, the European LeukemiaNet and the National Comprehensive Cancer Network (NCCN) have presented international guidelines on CML molecular approaches, especially those related to BCR::ABL1 expression. In this study, we show almost three years' worth of data regarding the clinical treatment of CML patients at the Erasto Gaertner Hospital, Curitiba, Brazil. These data primarily comprise 155 patients and 532 clinical samples. BCR::ABL1 quantification by a duplex-one-step RT-qPCR and ABL1 mutations detection were conducted. Furthermore, digital PCR for both BCR::ABL1 expression and ABL1 mutations were conducted in a sub-cohort. This manuscript describes and discusses the clinical importance and relevance of molecular biology testing in Brazilian CML patients, demonstrating its cost-effectiveness.
Asunto(s)
Proteínas de Fusión bcr-abl , Leucemia Mielógena Crónica BCR-ABL Positiva , Humanos , Brasil , Proteínas de Fusión bcr-abl/genética , Resistencia a Antineoplásicos/genética , Leucemia Mielógena Crónica BCR-ABL Positiva/diagnóstico , Leucemia Mielógena Crónica BCR-ABL Positiva/tratamiento farmacológico , Leucemia Mielógena Crónica BCR-ABL Positiva/genética , Mutación , Inhibidores de Proteínas Quinasas/farmacología , Translocación GenéticaRESUMEN
Use of the potent tyrosine kinase inhibitor imatinib as the first-line treatment in chronic myeloid leukemia (CML) has decreased mortality from 20% to 2%. Approximately 30% of CML patients experience imatinib resistance, however, largely because of point mutations in the kinase domain of the BCR-ABL1 fusion gene. The aim of this study was to use next-generation sequencing (NGS) to identify mutations related to imatinib resistance. The study included 22 patients diagnosed with CML and experiencing no clinical response to imatinib. Total RNA was used for cDNA synthesis, with amplification of a fragment encompassing the BCR-ABL1 kinase domain using a nested-PCR approach. Sanger and NGS were applied to detect genetic alterations. HaplotypeCaller was used for variant calling, and STAR-Fusion software was applied for fusion breakpoint identification. After sequencing analysis, F311I, F317L, and E450K mutations were detected respectively in three different participants, and in another two patients, single nucleotide variants in BCR (rs9608100, rs140506, rs16802) and ABL1 (rs35011138) were detected. Eleven patients carried e14a2 transcripts, nine had e13a2 transcripts, and both transcripts were identified in one patient. One patient had co-expression of e14a2 and e14a8 transcripts. The results identify candidate single nucleotide variants and co-expressed BCR-ABL1 transcripts in cellular resistance to imatinib.
Asunto(s)
Proteínas de Fusión bcr-abl , Leucemia Mielógena Crónica BCR-ABL Positiva , Humanos , Mesilato de Imatinib/uso terapéutico , Proteínas de Fusión bcr-abl/genética , Leucemia Mielógena Crónica BCR-ABL Positiva/tratamiento farmacológico , Leucemia Mielógena Crónica BCR-ABL Positiva/genética , Leucemia Mielógena Crónica BCR-ABL Positiva/diagnóstico , Mutación , Inhibidores de Proteínas Quinasas/uso terapéutico , Nucleótidos/uso terapéutico , Resistencia a Antineoplásicos/genéticaRESUMEN
BCR-ABL1 negative atypical chronic myeloid leukemia (aCML) is a rare type of myeloproliferative / myelodysplastic syndrome characterized by leukocytosis and proliferation of dysplastic neutrophilic precursors in the absence of positivity for the BCR-ABL1 fusion gene. We report a 66-year-old woman and a 57-year-old man with aCML, who initially presented with general malaise and weight loss, associated with anemia, thrombocytopenia, and leukocytosis with left shift and dysplasia in the neutrophil series. Both evolved unfavorably after admission and died a few days later due to multiple organ failure.
Asunto(s)
Leucemia Mielógena Crónica BCR-ABL Positiva , Leucemia Mieloide Crónica Atípica BCR-ABL Negativa , Trombocitopenia , Anciano , Femenino , Humanos , Leucemia Mielógena Crónica BCR-ABL Positiva/diagnóstico , Leucemia Mielógena Crónica BCR-ABL Positiva/genética , Leucemia Mieloide Crónica Atípica BCR-ABL Negativa/genética , Leucocitosis , Masculino , Persona de Mediana EdadRESUMEN
The present research refers to elaborating a new label-free electrochemical biosensor used to detect the BCR/ABL fusion gene. We used a hybrid nanocomposite composed of chitosan and zinc oxide nanoparticles (Chit-ZnONP) immobilized on a polypyrrole (PPy) film. DNA segments were covalently immobilized, allowing biomolecular recognition. Atomic force microscopy (AFM), cyclic voltammetry (CV), and electrochemical impedance spectroscopy (EIS) were used to evaluate the assembly stages of the biosensor. The biosensor's analytical performance was investigated using recombinant plasmids containing the target oncogene and clinical samples from patients with chronic myeloid leukemia (CML). A limit of detection (LOD) of 1.34 fM, limit of quantification (LOQ) of 4.08 fM, and sensitivity of 34.03 µA fM-1 cm2 were calculated for the BCR/ABL fusion oncogene. The sensing system exhibited high specificity, selectivity, and reproducibility with a standard deviation (SD) of 4.21%. Additionally, a linear response range was observed between 138.80 aM to 13.88 pM with a regression coefficient of 0.96. Also, the biosensor shows easy operationalization and fast analytical response, contributing to the early cancer diagnosis. The proposed nanostructured device is an alternative for the genetic identification BCR/ABL fusion gene.
Asunto(s)
Técnicas Biosensibles , Leucemia Mielógena Crónica BCR-ABL Positiva , Nanocompuestos , Técnicas Biosensibles/métodos , ADN/genética , Técnicas Electroquímicas/métodos , Humanos , Leucemia Mielógena Crónica BCR-ABL Positiva/diagnóstico , Leucemia Mielógena Crónica BCR-ABL Positiva/genética , Nanocompuestos/química , Polímeros/química , Pirroles , Reproducibilidad de los ResultadosRESUMEN
Chronic myelogenous leukemia (CML) is a myeloproliferative neoplasm that expresses the Philadelphia chromosome and constitutively activated Bcr-Abl tyrosine kinase in hematopoietic progenitor cells. Bcr-Abl tyrosine-kinase inhibitors (TKI) do not definitively cure all CML patients. The efficacy of TKI is reduced in CML patients in the blastic phase-the most severe phase of the disease-and resistance to this drug has emerged. There is limited knowledge on the underlying mechanisms of disease progression and resistance to TKI beyond BCR-ABL1, as well as on the impact of TKI treatment and disease progression on the metabolome of CML patients. The present study reports the metabolomic profiles of CML patients at different phases of the disease treated with TKI. The plasma metabolites from CML patients were analyzed using liquid chromatography, mass spectrometry, and bioinformatics. Distinct metabolic patterns were identified for CML patients at different phases of the disease and for those who were resistant to TKI. The lipid metabolism in CML patients at advanced phases and TKI-resistant patients is reprogrammed, as detected by analysis of metabolomic data. CML patients who were responsive and resistant to TKI therapy exhibited distinct enriched pathways. In addition, ceramide levels were higher and sphingomyelin levels were lower in resistant patients compared with control and CML groups. Taken together, the results here reported established metabolic profiles of CML patients who progressed to advanced phases of the disease and failed to respond to TKI therapy as well as patients in remission. In the future, an expanded study on CML metabolomics may provide new potential prognostic markers for disease progression and response to therapy.
Asunto(s)
Resistencia a Antineoplásicos , Leucemia Mielógena Crónica BCR-ABL Positiva , Biomarcadores , Progresión de la Enfermedad , Resistencia a Antineoplásicos/genética , Humanos , Leucemia Mielógena Crónica BCR-ABL Positiva/diagnóstico , Leucemia Mielógena Crónica BCR-ABL Positiva/tratamiento farmacológico , Leucemia Mielógena Crónica BCR-ABL Positiva/genética , Lípidos/uso terapéutico , Inhibidores de Proteínas Quinasas/farmacología , Inhibidores de Proteínas Quinasas/uso terapéuticoRESUMEN
BCR-ABL1 negative atypical chronic myeloid leukemia (aCML) is a rare type of myeloproliferative / myelodysplastic syndrome characterized by leukocytosis and proliferation of dysplastic neutrophilic precursors in the absence of positivity for the BCR-ABL1 fusion gene. We report a 66-year-old woman and a 57-year-old man with aCML, who initially presented with general malaise and weight loss, associated with anemia, thrombocytopenia, and leukocytosis with left shift and dysplasia in the neutrophil series. Both evolved unfavorably after admission and died a few days later due to multiple organ failure.
Asunto(s)
Humanos , Masculino , Femenino , Persona de Mediana Edad , Anciano , Trombocitopenia , Leucemia Mielógena Crónica BCR-ABL Positiva/diagnóstico , Leucemia Mielógena Crónica BCR-ABL Positiva/genética , Leucemia Mieloide Crónica Atípica BCR-ABL Negativa/genética , LeucocitosisRESUMEN
The detection of BCR-ABL1 mRNA transcripts is essential to molecular chronic myeloid leukemia (CML) diagnosis. In most cases, the RT-qPCR technique is performed as the gold standard diagnosis tool for clinical cases. However, this method requires expensive reagents and equipment, such as a real-time thermal cycler, probes and master mix. Consequently, the development and validation of simple and low-cost methods are essential for a rapid CML diagnosis in less specialized and equipped centers. In this study, we develop and demonstrate an accessible, rapid, and low-cost method using RT-LAMP for BCR-ABL1 detection in both cell lines and CML clinical samples, using fluorescent and colorimetric assays. Both methods demonstrated diagnostic specificity of 100% and while diagnostic sensitivity reaches more than 90% in samples with RT-qPCR cycle threshold above 31. The obtained data indicates that the proposed method here described is robust, specific and rapid approach for CML diagnosis with outstanding performance, especially for CML diagnostic procedure where present high BCR-ABL1 expression.
Asunto(s)
Colorimetría , Proteínas de Fusión bcr-abl/genética , Leucemia Mielógena Crónica BCR-ABL Positiva/diagnóstico , Técnicas de Diagnóstico Molecular , Técnicas de Amplificación de Ácido Nucleico , Línea Celular Tumoral , Proteínas de Fusión bcr-abl/metabolismo , Humanos , Leucemia Mielógena Crónica BCR-ABL Positiva/genética , ARN Mensajero/genética , ARN Mensajero/metabolismo , Espectrometría de FluorescenciaRESUMEN
This observational, multicenter study aimed to report the clinical evolution of COVID-19 in patients with chronic myeloid leukemia in Latin America. A total of 92 patients presented with COVID-19 between March and December 2020, 26% of whom were severe or critical. The median age at COVID-19 diagnosis was 48 years (22-79 years), 32% were 60 years or older, and 61% were male. Thirty-nine patients presented with at least one comorbidity (42.3%). Eighty-one patients recovered (88%), and 11 (11.9%) died from COVID-19. There was one case of reinfection. Patients with a major molecular response presented superior overall survival compared to patients with no major molecular response (91 vs. 61%, respectively; p = 0.004). Patients in treatment-free remission and receiving tyrosine kinase inhibitors showed higher survival rates than patients who underwent hematopoietic stem cell transplantation and those who did not receive tyrosine kinase inhibitors (100, 89, 50, and 33%, respectively; p < 0.001).
Asunto(s)
COVID-19 , Leucemia Mielógena Crónica BCR-ABL Positiva , Prueba de COVID-19 , Humanos , América Latina/epidemiología , Leucemia Mielógena Crónica BCR-ABL Positiva/diagnóstico , Leucemia Mielógena Crónica BCR-ABL Positiva/epidemiología , Leucemia Mielógena Crónica BCR-ABL Positiva/terapia , Masculino , SARS-CoV-2Asunto(s)
Proteínas de Fusión bcr-abl , Leucemia Mielógena Crónica BCR-ABL Positiva , ADN , Proteínas de Fusión bcr-abl/genética , Genómica , Humanos , Leucemia Mielógena Crónica BCR-ABL Positiva/diagnóstico , Leucemia Mielógena Crónica BCR-ABL Positiva/genética , Reacción en Cadena de la PolimerasaRESUMEN
O objetivo deste relato é apresentar o caso clínico de uma cadela, sem raça definida, com cinco anos de idade, diagnosticada com leucemia mieloide crônica (LMC). As leucemias são neoplasias malignas que se originam de células precursoras da medula óssea e as consequências podem ser trombocitopenia, anemia, leucocitose persistente e presença de células neoplásicas no sangue. O tratamento de escolha envolve o uso de inibidores de tirosina quinase, porém este não pode ser usado neste caso. Dessa forma a cadela recebeu diferentes protocolos quimioterápicos que incluíram inicialmente hidroxiureia, citarabina, doxorrubicina e prednisona. Devido a remissão parcial dos sinais clínicos e a resposta terapêutica pouco duradoura a essas medicações o protocolo foi alterado para quimioterapia metronômica com clorambucil. O uso desses quimioterápicos não foram eficazes em reduzir a leucocitose e controlar a anemia e trombocitopenia da paciente, devido a ocorrência do surgimento de células imaturas no sangue e resistência aos quimioterápicos. Na ausência da crise e da possibilidade do uso dos inibidores de tirosina quinase, a hidroxiureia permanece sendo o quimioterápico de eleição. O animal apresentou sobrevida de 210 dias, devido a leucocitose e anemia severas pouco responsivas ao protocolo terapêutico utilizado e o surgimento no hemograma de precursores neutrofilicos que ocorreu 46 dias
The aim of this report is to present the clinical case of a five-year-old mixed breed female dog diagnosed with chronic myeloid leukemia (CML). Leukemias are malignant neoplasms that originate from bone marrow precursor cells and the consequences can be thrombocytopenia, anemia, persistent leukocytosis and the presence of neoplastic cells in the blood. The treatment of choice involves the use of tyrosine kinase inhibitors, but it cannot be used in this case. Thus, the dog received different chemotherapy protocols that initially included hydroxyurea, cytarabine, doxorubicin and prednisone. Due to the partial remission of clinical signs and the short-term therapeutic response to these medications, the protocol was changed to metronomic chemotherapy with chlorambucil. The use of these chemotherapeutic agents was not effective in reducing leukocytosis and controlling the patients anemia and thrombocytopenia, due to the occurrence of immature cells in the blood and resistance to chemotherapeutic agents. In the absence of the crisis and the possibility of using tyrosine kinase inhibitors, hydroxyurea remains the chemotherapy of choice. The animal had a 210-day survival, due to severe leukocytosis and anemia, which were not responsive to the therapeutic protocol used and the appearance in the blood count of neutrophilic precursors that occurred 46 days after the beginning of hydroxyurea treatment.
Asunto(s)
Animales , Perros , Perros/anomalías , Leucemia Mielógena Crónica BCR-ABL Positiva/diagnóstico , Leucemia Mielógena Crónica BCR-ABL Positiva/terapia , Leucemia Mielógena Crónica BCR-ABL Positiva/veterinaria , Quimioterapia , Técnicas de Laboratorio ClínicoRESUMEN
Objectives The quantitation of BCR-ABL1 mRNA is mandatory for chronic myeloid leukemia (CML) patients, and RT-qPCR is the most extensively used method in testing laboratories worldwide. Nevertheless, substantial variation in RT-qPCR results makes inter-laboratory comparability hard. To facilitate inter-laboratory comparative assessment, an international scale (IS) for BCR-ABL1 was proposed. Methods The laboratory-specific conversion factor (CF) to the IS can be derived from the World Health Organization (WHO) genetic reference panel; however, this material is limited to the manufacturers to produce and calibrate secondary reference reagents. Therefore, we developed secondary reference calibrators, as lyophilized cellular material, aligned to the IS. Our purpose was both to re-evaluate the CF in 18 previously harmonized laboratories and to propagate the IS to new laboratories. Results Our field trial including 30 laboratories across Latin America showed that, after correction of raw BCR-ABL1/ABL1 ratios using CF, the relative mean bias was significantly reduced. We also performed a follow-up of participating laboratories by annually revalidating the process; our results support the need for continuous revalidation of CFs. All participating laboratories also received a calibrator to determine the limit of quantification (LOQ); 90% of them could reproducibly detect BCR-ABL1, indicating that these laboratories can report a consistent deep molecular response. In addition, aiming to investigate the variability of BCR-ABL1 measurements across different RNA inputs, we calculated PCR efficiency for each individual assay by using different amounts of RNA. Conclusions In conclusion, for the first time in Latin America, we have successfully organized a harmonization platform for BCR-ABL1 measurement that could be of immediate clinical benefit for monitoring the molecular response of patients in low-resource regions.
Asunto(s)
Proteínas de Fusión bcr-abl/genética , Leucemia Mielógena Crónica BCR-ABL Positiva/diagnóstico , Leucemia Mielógena Crónica BCR-ABL Positiva/genética , Calibración , Humanos , América Latina , Control de Calidad , ARN Mensajero/genética , Reacción en Cadena en Tiempo Real de la Polimerasa/métodos , Estándares de Referencia , Reproducibilidad de los ResultadosRESUMEN
O objetivo deste relato é apresentar o caso clínico de uma cadela, sem raça definida, com cinco anos de idade, diagnosticada com leucemia mieloide crônica (LMC). As leucemias são neoplasias malignas que se originam de células precursoras da medula óssea e as consequências podem ser trombocitopenia, anemia, leucocitose persistente e presença de células neoplásicas no sangue. O tratamento de escolha envolve o uso de inibidores de tirosina quinase, porém este não pode ser usado neste caso. Dessa forma a cadela recebeu diferentes protocolos quimioterápicos que incluíram inicialmente hidroxiureia, citarabina, doxorrubicina e prednisona. Devido a remissão parcial dos sinais clínicos e a resposta terapêutica pouco duradoura a essas medicações o protocolo foi alterado para quimioterapia metronômica com clorambucil. O uso desses quimioterápicos não foram eficazes em reduzir a leucocitose e controlar a anemia e trombocitopenia da paciente, devido a ocorrência do surgimento de células imaturas no sangue e resistência aos quimioterápicos. Na ausência da crise e da possibilidade do uso dos inibidores de tirosina quinase, a hidroxiureia permanece sendo o quimioterápico de eleição. O animal apresentou sobrevida de 210 dias, devido a leucocitose e anemia severas pouco responsivas ao protocolo terapêutico utilizado e o surgimento no hemograma de precursores neutrofilicos que ocorreu 46 dias(AU)
The aim of this report is to present the clinical case of a five-year-old mixed breed female dog diagnosed with chronic myeloid leukemia (CML). Leukemias are malignant neoplasms that originate from bone marrow precursor cells and the consequences can be thrombocytopenia, anemia, persistent leukocytosis and the presence of neoplastic cells in the blood. The treatment of choice involves the use of tyrosine kinase inhibitors, but it cannot be used in this case. Thus, the dog received different chemotherapy protocols that initially included hydroxyurea, cytarabine, doxorubicin and prednisone. Due to the partial remission of clinical signs and the short-term therapeutic response to these medications, the protocol was changed to metronomic chemotherapy with chlorambucil. The use of these chemotherapeutic agents was not effective in reducing leukocytosis and controlling the patients anemia and thrombocytopenia, due to the occurrence of immature cells in the blood and resistance to chemotherapeutic agents. In the absence of the crisis and the possibility of using tyrosine kinase inhibitors, hydroxyurea remains the chemotherapy of choice. The animal had a 210-day survival, due to severe leukocytosis and anemia, which were not responsive to the therapeutic protocol used and the appearance in the blood count of neutrophilic precursors that occurred 46 days after the beginning of hydroxyurea treatment.(AU)
Asunto(s)
Animales , Perros , Perros/anomalías , Leucemia Mielógena Crónica BCR-ABL Positiva/diagnóstico , Leucemia Mielógena Crónica BCR-ABL Positiva/terapia , Leucemia Mielógena Crónica BCR-ABL Positiva/veterinaria , Técnicas de Laboratorio Clínico , QuimioterapiaRESUMEN
CONTEXTO: O diagnóstico das leucemias Ph+ é feito pela demonstração da presença do cromossomo Philadelphia (22q-) com translocação t (9;22) (q34; q11) ou rearranjo BCR-ABL, identificado por citogenética (já disponível no SUS), ISH ou método molecular (RT-PCR), em pacientes com leucocitose (e ocasionalmente trombocitemia) persistente. Além disso, o Leukemianet recomenda que RT-qPCR seja utilizado no monitoramento do tratamento; nos casos de falha ou progressão da doença, para a detecção de mutações; e nos casos que requerem maior atenção. PERGUNTA: Qual a acurácia diagnóstica da Reação em Cadeia da Polimerase Transcriptase Reversa (RT-PCR) qualitativa e quantitativa (RT-qPCR) e da Hibridização in situ (ISH) para o diagnóstico e monitoramento das leucemias Philadelphia positivo em adultos e crianças/adolescentes? TECNOLOGIA: Hibridização in situ (ISH), Reação em Cadeia da polimerase por transcriptase reversa qualitativa (RT-PCR) e Reação em Cadeia da polimerase por transcriptase reversa quantitativa (RT-qPCR). EVIDÊNCIAS CIENTÍFICAS: As bases Medline (via Pubmed) e Embase foram pesquisadas e onze estudos foram elegíveis para responder a pergunta de pesquisa. Esses estudos foram agrupados de acordo com os respectivos testes índice e referência. Nenhum dos estudos incluídos foi realizado em populações com suspeita de leucemias Ph+ (triagem). Todos os estudos incluem pacientes com diagnóstico confirmado de leucemias ou outras doenças hematológicas, proliferativas ou não, e avaliam a acurácia com base na detecção dos casos positivos (detecção do transcrito BCR-ABL) e negativos (doenças hematológicas não relacionadas ao BCR-ABL, leucemias Ph- ou indivíduos sadios). Alguns estudos inclusive avaliam a capacidade de detecção e quantificação de transcritos de um teste após a terapia com imatinibe, interferon ou transplante de medula óssea (TMO) (avaliação da resposta molecular). De modo geral, houve boa acurácia entre as técnicas de ISH e RT-PCR (quali e quantitativo), RT-qPCR e RT-PCR qualitativo, tipos diferentes de RT-PCR e RT-qPCR conduzido em amostras de sangue periférico e medula óssea (MO). A maioria dos resultados de sensibilidade e especificidade foi superior a 80%. AVALIAÇÃO DE IMPACTO ORÇAMENTÁRIO (AIO): O cenário simulado com o menor preço de mercado apresentou o menor impacto orçamentário. No contexto do diagnóstico, esses custos poderiam variar entre R$ 1.313.872,46 e R$ 2.920.036,56, a depender da combinação da frequência de uso das tecnologias, no período entre 2019 e 2023. No contexto do monitoramento, no qual apenas a técnica de RT-PCR quantitativo é utilizada, os valores variaram de R$ 67.281.142,19, para 4 avaliações/ano, até R$ 16.820.285,55, para uma avaliação/ano. CONSIDERAÇÕES FINAIS: O diagnóstico de leucemias Ph+ típica é simples e consiste em documentar, na presença de leucocitose persistente (ou ocasionalmente trombocitose), a presença da anormalidade cromossômica Filadélfia (Ph), o t (9; 22) (q34; q11), por citogenética de rotina, ou anormalidades moleculares BCR-ABL1 relacionadas ao Ph, por hibridização fluorescente in situ (FISH) ou por estudos moleculares. Estudos de acurácia diagnóstica evidenciam bons resultados de sensibilidade e especificidade para os testes ISH e RT-PCR (qualitativo e quantitativo). Ademais, um desses estudos mostra que existe boa correlação entre os resultados obtidos por RT-qPCR em amostras de sangue periférico e em MO. Os testes diagnósticos pleiteados nesse relatório eram anteriormente fornecidos aos pacientes com LMC e LLA Ph+ pelas empresas fabricantes dos inibidores de tirosinoquinase, fato que não acontece atualmente e que pode deixar os usuários sem a integralidade assistencial necessária. Esse relatório pleiteia, portanto, a ampliação do uso de ISH e RT-PCR para LMC e LLA Ph+, no âmbito do SUS, haja vista que, atualmente, esses procedimentos são fornecidos apenas para portadores de doenças raras, que não incluem as leucemias. A AIO mais atrativa considerou os menores valores praticados no mercado. No contexto do diagnóstico, houve variação entre R$ 1.313.872,46 e R$ 2.920.036,56, no período entre 2019 e 2013. No contexto do monitoramento, os valores variaram de R$ 67.281.142,19, para 4 avaliações/ano, até R$ 16.820.285,55, para uma avaliação/ano. RECOMENDAÇÃO PRELIMINAR DA CONITEC: A Conitec, em sua 78ª reunião ordinária, realizada no dia 05 e 06 de junho de 2019, recomendou a incorporação a incorporação dos testes diagnósticos ISH e RT-PCR (qualitativo e quantitativo), para o diagnóstico e o monitoramento das leucemias Ph+ (LMC e LLA Ph+) em adultos e em crianças/adolescentes. A matéria foi disponibilizada para consulta pública. CONSULTA PÚBLICA: Foram apresentadas as características das 641 (seiscentas e quarenta e uma) contribuições recebidas, das quais 400 foram de experiência e opinião e 241 técnico científicas. Do total de contribuições, 99% dos participantes concordaram totalmente com a recomendação preliminar da Comissão Nacional de Incorporação de Tecnologias no Sistema Único de Saúde. RECOMENDAÇÃO FINAL DA CONITEC: Os membros presentes deliberaram, por unanimidade, recomendar a incorporação, ao SUS, dos testes de reação em cadeia da polimerase transcriptase reversa (RT-PCR) qualitativa e quantitativa (RT-qPCR) e hibridização in situ (ISH) para o diagnóstico e monitoramento da leucemia mieloide crônica (LMC) e da leucemia linfoblástica aguda cromossoma Philadelphia positivo (LLA Ph+). Foi assinado o Registro de Deliberação nº 464/2019. DECISÃO: Incorporar a reação em cadeia da polimerase - transcriptase reversa (RT-PCR) qualitativa e quantitativa (RT-qPCR) e hibridização in situ (ISH) para o diagnóstico e monitoramento da leucemia mieloide crônica (LMC) e da leucemia linfoblástica aguda cromossoma Philadelphia positivo (LLA Ph+), no âmbito do Sistema Único de Saúde - SUS. Dada pela Portaria nº 57, publicada no Diário Oficial da União nº 224, seção 1, página 79, em 20 de novembro de 2019.
Asunto(s)
Humanos , Leucemia Mielógena Crónica BCR-ABL Positiva/diagnóstico , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa/instrumentación , Leucemia-Linfoma Linfoblástico de Células Precursoras/diagnóstico , Evaluación de la Tecnología Biomédica , Sistema Único de Salud , Brasil , Análisis Costo-Beneficio/economíaRESUMEN
This study evaluated the effectiveness of thermocycler temperature control, considering the influence of other determinant factors for the optimization of PCR. The reduction in the number of repeated PCR tests, applied in the diagnosis and prognosis of chronic myeloid leukemia at the National Cancer Institute in Brazil, was used as a measure of effectiveness. This indicator was evaluated using samples obtained before and after the temperature control in the wells of the thermocyclers. There was a reduction of 18.9% in the number of repeated exams in the second sample. A structured interview with laboratory staff indicated that there was no change in the other determinant factors.
Asunto(s)
Reacción en Cadena de la Polimerasa/métodos , Brasil , Cartilla de ADN/química , Humanos , Leucemia Mielógena Crónica BCR-ABL Positiva/diagnóstico , TemperaturaAsunto(s)
Crisis Blástica/complicaciones , Médula Ósea/patología , Leucemia Mielógena Crónica BCR-ABL Positiva/complicaciones , Enfermedades del Nervio Óptico/etiología , Nervio Óptico/patología , Agudeza Visual , Crisis Blástica/diagnóstico , Humanos , Leucemia Mielógena Crónica BCR-ABL Positiva/diagnóstico , Imagen por Resonancia Magnética , Masculino , Persona de Mediana Edad , Enfermedades del Nervio Óptico/diagnóstico , Microscopía con Lámpara de Hendidura , Pruebas del Campo VisualRESUMEN
A 12-year-old Rottweiler dog was presented with a history of prostration, weight loss and hyporexia for six months. Based on complete blood tests (hematological and biochemical analyses), bone marrow examination and imaging analysis, a diagnosis of chronic myeloid leukemia was made. Treatment with hydroxyurea at a dosage of 18â¯mg/kg twice daily was not effective in controlling the high count of white blood cells. Furthermore, after 35 days of hydroxyurea treatment, the animal developed onycholysis, with sloughing of the claws of the left pelvic and left thoracic limbs and exposure of the distal phalanx. Interruption of the medication was implemented, with clinical healing of the ungual lesions observed three months after initiation of the drug. White blood cells returned to normal after using cyclophosphamide. Currently, the animal is in complete remission, having a disease-free interval of 575 days without chemotherapy. To the authors' knowledge, this is the first report of hydroxyurea-induced onycholysis within a short-term period in a dog diagnosed with chronic myeloid leukemia.