El rol del laboratorio en los hallazgos de intoxicación y exposición a monóxido de carbono / The role of the laboratory in findings of carbon monoxide poisoning and exposure

El monóxido de carbono es un gas altamente tóxico que se origina principalmente por la combustión incompleta de combustibles fósiles. La intoxicación presenta síntomas inespecíficos que solapan otras patologías y por lo tanto es indispensable la confirmación mediante la medición de la carboxihemoglobina en sangre. El laboratorio incorporó la determinación en el informe del estado ácido base a partir de octubre del 2018, debido a que previamente el médico debía solicitarla frente a la sospecha de una intoxicación. El objetivo del trabajo fue evaluar si esta medida implementada por el laboratorio contribuyó a mejorar el diagnóstico de intoxicación por CO, analizar las características de los pacientes con COHb mayor o igual a 5% y definir un valor de reporte inmediato para la COHb. El 46% de los casos con COHb mayor o igual a 5% no se relacionaban con una intoxicación y/o exposición a CO. De los casos de intoxicación se encontró que el 77% fueron diagnosticados a partir de la sospecha médica y un 23% por hallazgo del laboratorio. Se concluyó que es de mucha utilidad el rol del laboratorio en detectar aquellos casos que no fueron evidentes clínicamente. Existen ciertas patologías como las oncológicas o la enfermedad de Wilson donde se vieron valores elevados de COHb sin presentar intoxicación y se definió finalmente, como valor de reporte inmediato 7% para la COHb. (AU)

Carbon monoxide is a highly toxic gas that originates mainly from incomplete combustion of fossil fuels. Intoxication causes nonspecific symptoms that overlap with other conditions and, therefore, confirmation by measuring blood carboxyhemoglobin is essential. The laboratory incorporated the measurement in the acid-base status report as of October 2018, as it was previously required to be requested by the physician in case of suspected intoxication. The aim of this study was to evaluate whether this measure implemented by the laboratory contributed to the improvement of the diagnosis of CO intoxication, to analyze the characteristics of patients with COHb greater than or equal to 5% and to define an immediate reporting value for COHb. Overall, 46% of the cases with COHb greater than or equal to 5% were not related to CO poisoning and/or exposure. Of the cases of intoxication, 77% were diagnosed based on medical suspicion and 23% on laboratory findings. It was concluded that the laboratory has a useful role in detecting cases that were not clinically evident. There are certain diseases including different types of cancer or Wilson's disease where elevated COHb values were seen without intoxication and finally, 7% for COHb was defined as the immediate reporting value (AU)

Una experiencia de siete años en el laboratorio de Micología del Hospital de Pediatría Juan P. Garrahan / A seven-year experience in the Mycology Laboratory of Hospital de Pediatría Juan P. Garrahan

En noviembre del año 2015 nos incorporamos al Laboratorio de Micología del Servicio de Microbiología del Hospital Garrahan. En este breve resumen queremos compartir los avances logrados a través de nuestra experiencia durante siete años de trabajo profesional. Debido a los diagnósticos realizados y su complejidad, consideramos que el Hospital Garrahan, sus pacientes y la comunidad toda necesitan contar con un laboratorio de Micología que responda a sus necesidades. Creemos haber iniciado un camino que esperamos continúe y culmine con la creación de la Unidad de Micología (AU)

In November 2015 we joined the Mycology Laboratory of the Microbiology Service of the Hospital Garrahan. In this brief summary we want to share the advances achieved through our experience during seven years of professional work. Due to the diagnosis made and their complexity, we believe that the Hospital Garrahan, its patients and the entire community, need to have a Mycology laboratory that responds to their requirements. We believe we have started a path that we hope will continue and culminate with the creation of the Mycology Unit (AU)

Methylene Blue for the Treatment of Health Conditions: a Scoping Review

HIGHLIGHTS This scoping review summarizes the findings of clinical trials using methylene blue (MB) for the treatment of various health conditions. This research method allowed mapping main findings, clarifying research topics, and identifying gaps in the literature.

Abstract studies evaluating effective drugs for health conditions are of crucial importance for public health. Methylene blue (MB) is an accessible synthetic drug that presents low toxicity and has been used in several health areas due to its effectiveness. Objective: this scoping review aims to provide a comprehensive overview of relevant research regarding the use of MB for the treatment of health conditions. Methods: a five-stage framework Arksey and O'maley scoping review was conducted. The literature was searched in Cochrane Library database using Mesh term "methylene blue". Data were collected by two independent reviewers and submitted to descriptive synthesis. Results: The search resulted in 429 records, from which 16 were included after exclusion criteria were applied. The therapeutic use of MB was identified for acute conditions (malaria and septic shock), chronic conditions (discogenic back pain, bipolar disorder, refractory neuropathic pain, and post-traumatic stress disorder), and postoperative care (vasoplegic syndrome, and pain after haemorrhoidectomy, lumbar discectomy, and traumatic thoracolumbar fixation). Conclusion: there is much evidence emerging from clinical trials about the therapeutic use of MB for acute, chronic, and postoperative conditions; however, many gaps were identified, which open further avenues for future research.

Association between soluble biomarkers - microbial translocation, inflammation and cardiovascular risk in HIV- infected individuals: a systematic review

Microbial translocation is associated with the increased risk of cardiovascular disease in HIV-infected individuals. There is scarce information regarding the possible associations between the biomarkers of microbial translocation, inflammation and cardiovascular risk that can be evaluated in clinical laboratories using plasma or serum samples. This systematic review was conducted according to the PRISMA protocol in order to verify the most used soluble biomarkers of microbial translocation, inflammation and cardiovascular risk, as well as possible associations between them, in HIV-infected individuals. A search was performed using the Medline, Scopus and Web of Science databases to identify existing studies regarding the relationship between microbial translocation biomarkers, inflammation and cardiovascular risk in HIV-infected patients. Eleven articles that presented soluble biomarkers of microbial translocation (LPS, rDNA, sCD14, LBP and EndoCAb) were selected. The most frequently evaluated soluble biomarker was sCD14, followed by LPS; the latter were associated with some lipid profile parameters. This systematic review considered soluble blood biomarkers that can be utilized in laboratory diagnosis. The aim was to identify the interconnection between microbial translocation, inflammation and cardiovascular risk. Despite the fact that a large number of inflammation and cardiovascular risk biomarkers have been previously reported, it was noted that important markers involved in the pathophysiology of cardiovascular diseases need to be included in future research.

Respuesta del score RIPASA versus el histopatológico como predictor diagnóstico de apendicitis aguda en el Hospital Vicente Corral Moscoso año 2018 / Response of the RIPASA score versus the histopathology as a diagnostic predictor of acute appendicitis at the Vicente Corral Moscoso Hospital in 2018

Antecedentes: la Apendicitis Aguda (A.A) es una urgencia quirúrgica que requiere un diagnóstico y tratamiento oportuno. Muchas veces puede ser un gran reto para el cirujano por su relación con otras patologías, de allí la importancia de precisar su diagnóstico. Objetivo: validar la precisión diagnóstica del score RIPASA en apendicitis aguda comparándolo con el examen histopatológico. Metodología: se realizó un estudio de validación de pruebas diagnósticas con la información de 300 historias clínicas de pacientes apendicectomizados, atendidos en el Hospital Vicente Corral Moscoso durante el año 2018. Se evaluó mediante el score RIPASA al ingreso y se comparó con los resultados de histopatología como prueba gold standar. Se calculó la sensibilidad, especificidad, valor predictivo positivo, valor predictivo negativo, además se obtuvo Odds Ratio con su IC al 95% para establecer la validez predictiva de esta escala. Resultados: la media de edad fue de 32 años ± 13,7 (DS), más de la mitad fueron: sexo femenino 52%, área urbana 74.7% y bachillerato 62.7%, predominó la etnia mestiza 99.7%. Hubo una asociación de riesgo entre una alta probabilidad de apendicitis según el score RIPASA con A.A (OR 96,36; IC95%: 16,03­578,68; p= 0,000). El score RIPASA tiene una sensibilidad de 98.97%, especificidad 50.0%, VPP 98.63%, VPN 57,14%, RVP 1.98% Y RVN 0.2%. Conclusiones: RIPASA tiene alta probabilidad para detectar a personas con apendicitis aguda, pero no así a las sanas, por lo cual se requiere continuar con más estudios para establecer tal validez

Background: Acute Appendicitis (A.A) is a surgical emergency that requires timely diagnosis and treatment. Many times it can be a great challenge for the surgeon because of its relationship with other pathologies, hence the importance of specifying its diagnosis. Objective: To validate the diagnostic accuracy of the RIPASA score in acute appendicitis by comparing it with the histopathological examination. Methodology: A study of validation of diagnostic tests was carried out with the information of 300 medical records of appendectomized patients, treated at the Vicente Corral Moscoso Hospital during 2018. It was evaluated by means of the RIPASA score at admission and compared with the histopathology results. as gold standard test. Sensitivity, specificity, positive predictive value, negative predictive value were calculated, in addition Odds Ratio was obtained with its 95% CI to establish the predictive validity of this scale. Results: The mean age was 32 years ± 13.7 (SD), more than half were: female 52%, urban area 74.7% and high school 62.7%, mixed race ethnicity predominated 99.7%. There was a risk association between a high probability of appendicitis according to the RIPASA score with A.A (OR 96.36; 95% CI: 16.03­578.68; p = 0.000). The RIPASA score has a sensitivity of 98.97%, specificity 50.0%, PPV 98.63%, NPV 57.14%, RVP 1.8% and RVN 0.2% Conclusions: RIPASA has a high probability of detecting people with acute appendicitis, but not healthy ones, so it is necessary to continue with more studies to find such validity

Laboratorio de diagnóstico clínico y/o de vigilancia epidemiológica: norma técnica No. 22-2019-DRACES / Laboratory for clinical diagnosis and / or epidemiological surveillance: technical standard No. 22-2019-DRACES

La tinta del documento está bastante opaca. DRACES [Departamento de Regulación, Acreditación y Control de Establecimientos de Salud] Este documento tiene como objeto: "la regulación, autorización y control de los laboratorios de diagnóstico clínico y/o de vigilancia epidemiológica, en concordancia con el Reglamento para la Regulación, Autorización, Acreditación y Control de Establecimientos de Atención para la Salud, Acuerdo Gubernativo 376-2007." Es de carácter obligatorio, por lo que se aplica tanto al sector público, privado, social o subsector de la seguridad social, en todo el territorio nacional. Contiene además, las definiciones de los conceptos relacionados al tema principal, además de la infraestructura que deberá tener cada clínica, incluidos el equipo y recurso humano y técnico. En el capítulo II, incluye una clasificación del nivel de laboratorios, describiendo sus características, servicios, horarios, materiales técnicos y equipos.

Resistência aos compostos de amônio quaternário (QACs) de uso doméstico e hospitalar em patógenos prioritários multirresistentes / Resistance to quaternary ammonium compounds (QACs) for domestic and hospital use in multiresistant priority pathogens

Compostos de amônio quaternário (QACs) têm sido amplamente utilizados como desinfetantes e antissépticos, sendo essenciais na prevenção e controle de infecções bacterianas na medicina humana e veterinária. Embora patógenos prioritários multirresistentes têm sido muito bem caracterizados quanto ao perfil de suscetibilidade e contexto genético da resistência aos antibióticos, dados de resistência aos QACs são limitados. Assim, o objetivo do presente estudo foi avaliar a atividade in vitro dos QACs de uso doméstico e hospitalar [cloreto de benzalcônio (BAC), cloreto de cetilpiridinio (CPC) e brometo de cetiltrimetilamônio (CTAB)], contra patógenos prioritários multirresistentes, identificando os principais genes de resistência associados. Foram estudadas 100 cepas multirresistentes previamente sequenciados usando as plataformas Illumina MiSeq e NextSeq representativas de diferentes hospedeiros (humanos e animais) e fontes (ambientes e alimentos). As cepas foram identificadas como Klebsiella pneumoniae (n= 24), Escherichia coli (n= 30); Pseudomonas aeruginosa (n= 10), Enterobacter spp, (n= 8), Acinetobacter baumannii (n= 11) e Salmonella spp. (n= 17). Genes de resistência aos QACs foram identificados in silico através do alinhamento dos contigs obtidos de cada cepa sequenciada com genes de referência obtidos do GenBank, utilizando o programa Geneious versão 8 (Biomatters Ltd). A identidade de cada gene foi analisada utilizando o programa BLASTx, no qual um critério baseado em ≥90% identidade resultou na identificação dos genes mdfA (77%), qacE (44%), qacEΔ1 (43%), sugE(c) (29%), emrE (21%), qacA (19%), sugE(p) (5%), qacF (7%), qacH (7%) e qacL (7%) em 85 cepas; enquanto que 15 cepas não possuíam nenhum gene de resistência aos QACs. A concentração inibitória mínima (CIM) dos QACs para as 100 cepas foi determinada pelo método de microdiluição em caldo. Os resultados sugeriram que a resistência em patógenos prioritários circulando na interface humano-ambiente-animal não é restrita aos antibióticos, uma vez que a elevada ocorrência de genes qacE, qacEΔ1 e mdfA poderia estar associada com uma redução da suscetibilidade para QACs. Consequentemente, a resistência aos QACs poderia também contribuir para a persistência e adaptação destes patógenos nos seres humanos e outros animais, assim como em ambientes impactados antropogenicamente

Quaternary ammonium compounds (QACs) have been widely used as disinfectants and antiseptics, being applied as essential compounds in the prevention and control of bacterial infections in human-and veterinary hospital medicine. Although multiresistant priority pathogens have been well characterized with respect to their susceptibility profile and their genetic context of resistance for antibiotics, studies of resistance to QACs are limited. Thus, the objective of the present study was to evaluate the in vitro activity of QACs [(benzalkonium chloride (BAC), cetylpyridinium chloride (CPC) and cetyltrimethylammonium bromide (CTAB)] for household and hospital use against multiresistant priority pathogens, identifying the main resistance genes associated. A hundred multiresistant isolates (previously sequenced using the Illumina MiSeq and NextSeq platforms), representative of different hosts (humans and animals) and sources (environment and food) were studied. Isolates were identified as Klebsiella pneumoniae (n=24), Escherichia coli (n=30), Pseudomonas aeruginosa (n=10), Enterobacter spp. (n=8), Acinetobacter baumannii (n=11) and Salmonella spp. (n=17). In silico analysis for identification of genes conferring resistance to QACs were performed by aligning the contigs obtained from the strains with reference genes deposited in GenBank, using the Geneious version program (Biomatters Ltd). Similarities were analyzed using the BLASTx online program, considering the alignment criteria based on ≥ 90% identity. The result of these analysis revealed the presence of the following QAC genes: mdfA (77%), qacE (44%), qacEΔ1 (43%), sugE(c) (29%), emrE (21%), qacA (19%), sugE (p) (5%), qacF (7%), qacH (7%) e qacL (7%); while 15 strains showed no resistance genes for QACs. Determination of QACs minimum inhibitory concentration (MIC) for the 100 isolates, by the broth microdilution method. These results suggest that resistance to QACs in priority pathogens, circulating at the human-environment-animal interface, is not restricted to antibiotics, since the high occurrence of genes qacE, qacEΔ1 and mdfA were associated with a reduced susceptibility to QACs. Consequently, resistance to QACs could also contribute to the persistence and adaptation of these pathogens in humans and othes animals, as well as in anthropogenically impacted environments

Modelo de gestión impuesto por la transformación tecnológica en los laboratorios de análisis clínicos. Impacto en el Hospital Garrahan a treinta años de historia / Management model imposed by the technological transformation in clinical analysis laboratories. Impact at Hospital Garrahan on thirty years of history

Las actividades del laboratorio de análisis clínicos están fuertemente identificadas con el ritmo del cambio tecnológico. En los últimos treinta años se vive un cambio de época que afecta la cultura y la experiencia humana en todos sus aspectos. Este cambio modifico el modelo de producción. En el laboratorio ha incorporado tecnología que combina la química, la robótica, la óptica y la informática, y se ha impuesto como premisa externa de cambio un modelo de gestión global que afecta la forma de trabajo, la gestión y el rol de los profesionales (bioquímicos y técnicos). El hospital Garrahan inscribe su historia dentro de este periodo histórico y el proceso de cambio ha generado y genera incertidumbre, resistencia y adecuaciones al nuevo paradigma impuesto. Nos fijamos como objetivo analizar el impacto de este cambio sobre la gestión de procesos del laboratorio de nuestro hospital. Comprobamos que la demanda del laboratorio se incrementó al ritmo del crecimiento de consultas y egresos de pacientes, y de como este aumento demando adecuaciones de gestión, modificaciones arquitectónicas, incorporación de tecnologías (algunas emergentes), aumento en la trazabilidad de muestras y resultados, y mejoras en la seguridad del paciente en todos sus aspectos. Describimos el nuevo paradigma, sus ventajas, las adecuaciones hechas y los tiempos en que se fueron realizados. Concluimos sugiriendo un rol para los profesionales del laboratorio en función del paradigma en curso

Activities at the laboratory of clinical analysis are closely related to the pace of technological change. Over the past 30 years there has been a change of times affecting human culture and experience in all its aspects. This change has modified the model of production. The laboratory has incorporated technology combining chemistry, robotics, and optics, as well as information technology, and the premise of a global management model has been imposed affecting the way of working, administration, and the role of professionals (biochemists and technicians). Garrahan hospital has written its own history in this historical period and process of change has produced uncertainty, resistance as well as adaptation to this new paradigm. Our aim has been to analyze the impact of this change on the management of processes of the laboratory of our hospital. We have observed that the demand of the laboratory has increased at the same pace as the increase of patient visits and discharges. This increase has required modifications in the management and facilities, incorporation of new technologies (some of them state of the art), improved traceability of the samples and results, and improvements in patient safety in all aspects. Here we describe the new paradigm, its advantages, adaptations made, and improved times introduced. In our conclusions, we consider the new role for laboratory professionals in this paradigm.

Survey of Clinical Laboratory Practices for 2015 Middle East Respiratory Syndrome Coronavirus Outbreak in the Republic of Korea

BACKGROUND: It is crucial to understand the current status of clinical laboratory practices for the largest outbreak of Middle East respiratory syndrome coronavirus (MERS-CoV) infections in the Republic of Korea to be well prepared for future emerging infectious diseases. METHODS: We conducted a survey of 49 clinical laboratories in medical institutions and referral medical laboratories. A short questionnaire to survey clinical laboratory practices relating to MERS-CoV diagnostic testing was sent by email to the directors and clinical pathologists in charge of the clinical laboratories performing MERS-CoV testing. The survey focused on testing volume, reporting of results, resources, and laboratory safety. RESULTS: A total of 40 clinical laboratories responded to the survey. A total of 27,009 MERS-CoV real-time reverse transcription PCR (rRT-PCR) tests were performed. Most of the specimens were sputum (73.5%). The median turnaround time (TAT) was 5.29 hr (first and third quartile, 4.11 and 7.48 hr) in 26 medical institutions. The median TAT of more than a half of the laboratories (57.7%) was less than 6 hr. Many laboratories were able to perform tests throughout the whole week. Laboratory biosafety preparedness included class II biosafety cabinets (100%); separated pre-PCR, PCR, and post-PCR rooms (88.6%); negative pressure pretreatment rooms (48.6%); and negative pressure sputum collection rooms (20.0%). CONCLUSIONS: Clinical laboratories were able to quickly expand their diagnostic capacity in response to the 2015 MERS-CoV outbreak. Our results show that clinical laboratories play an important role in the maintenance and enhancement of laboratory response in preparation for future emerging infections.

Hermenéutica-dialéctica en la práctica clínica neurológica / Dialectic-hermeneutics in clinical practice neurology

Hablar de la hermenéutica-dialéctica, desde el enfoque de Miguel Martínez Migueles, implica abordar algunos antecedentes históricos de este método, la manera en la que es utilizado en la investigación tradicional, en las ciencias jurídicas y humanas, a través de la observación, y cómo puede ser utilizado en la práctica clínica para el diseño de un instrumento de valoración neurológica para los profesionales de enfermería.

Speaking of Dialectic-Hermeneutics, since the focus of Miguel Martinez Michaels is addressing some historical background of this method and how it is used in traditional research in legal sciences and humanities through observation and how it can be used in practice clinic for the design of a neurological assessment tool for nurses.

Laboratory diagnosis of chikungunya virus: Do we really need it.

Chikungunya (CHIK) fever is a re-emerging Aedes mosquito-transmitted viral disease caused by CHIK virus belonging to the Togaviridae family of genus Alphavirus. The disease is almost self-limiting, occurs with characteristic triad of sudden onset fever, rash and arthritis. During the recent outbreak CHIKV was also found to cause long-term arthralgia, severe neurological disease and even fatalities. Although there are no antiviral or vaccines available for CHIKV, still there are several advantages to diagnose the infection. The present article provides an overview of various diagnostic modalities available and its significance by searching PubMed MeSH terms "Chikungunya virus" and "Diagnosis" for recent articles. The gold standard of CHIKV diagnosis is culture, yet requires facilities and skills. Highly sensitive and specific PCR assays for CHIKV have been developed, but the reagents and equipment are costly for widespread use. Serological diagnosis by detecting IgM antibody is most widely used as it is relatively cheaper and easier to perform. Disadvantages of antibody testing are cross-reactivity with other alpha viruses, cannot differentiate between recent past and acute infection, and its sensitivity varies in clinical settings. When tested for diagnosing acute CHIKV disease, sensitivities were just 4 to 22% and after 1 week rose to more than 80%. As most acutely infected patients seek medical attention within the first few days of illness, the ideal test should detect RNA or antigen. Therefore, the more realistic aim would be to develop a reliable antigen detection assay that could be used in rural areas, where CHIKV infection often occurs.

Complicações diagnósticas na fibrose cística: métodos de rotina e novas alternativas / Complications in cystic fibrosis diagnostic: routine methods and new alternatives

A Fibrose Cística (FC) é uma doença grave presente em nosso meio. Apesar dos avanços nos métodos diagnósticos e no tratamento do paciente, existem inúmeros casos que ainda apresentam dificuldades para a definição desta doença. Os métodos mais comuns de diagnóstico são o teste de suor e análise de mutação para confirmação. Porém uma grande parcela dos pacientes com FC é portador de uma mutação não identificada e devem ser diagnosticados por várias outras medidas de disfunções orgânicas relatadas nesse artigo. O presente trabalho teve por objetivo descrever os principais meios de diagnosticar fibrose cística e a importância de uma definição precoce para que melhores resultados terapêuticos sejam atingidos.

Cystic Fibrosis (CF) is a several disease present in our midst despite the advances in diagnostic methods and patient's treatment. Despite the improvement there are still many cases that have complicated and unclear diagnosis. There are several ways to diagnosis cystic fibrosis, being more common the sweat test and mutation analysis to confirm. However, a large proportion of CF patients has a mutation that cannot be identified. These patients should be investigated by several other measures of organ dysfunction that this article reports. This study aimed to describe the principal means of diagnosing cystic fibrosis and the importance of an appropriate definition as soon as possible for better therapeutic results.

Desenvolvimento de imunoensaios e biossensores para determinação de LDL eletronegativa / Development of immunoassays and biosensors for electronegative LDL quantification

A lipoproteína de baixa densidade eletronegativa (LDL-) é um importante antígeno envolvido na patogênese da aterosclerose. A LDL(-) provoca resposta inflamatória e imunológica, levando à produção de autoanticorpos anti-LDL(-) e a formação subsequente de imunocomplexos (IC-LDL-), os quais também contribuem com o processo aterosclerótico. Diante disso, este trabalho teve como objetivo o desenvolvimento e validação de imunoensaios para quantificação plasmática de LDL(-), anti-LDL(-) e IC-LDL(-), assim como o desenvolvimento de ferramentas aplicáveis em um biossensor para LDL(-). Após a padronização de cada ELISA, foram avaliadas as características de desempenho dos métodos: limites de detecção (LD) e quantificação (LQ), precisão intra e inter-ensaios, exatidão, linearidade de diluição e interferentes. Os LD e LQ do ELISA para LDL(-) foram 0,423 mg/L e 0,517 mg/L de LDL(-), respectivamente. As concentrações plasmáticas de LDL(-) apresentaram linearidade quando os plasmas foram diluídos 1:1000, 1:2000, 1:4000 e 1:8000. Os LD e LQ do ELISA para anti-LDL(-) foram 0,0028 mg/L e 0,0032 mg/L de anti-LDL(-), respectivamente. Os plasmas apresentaram linearidade na diluição quando diluídos 1:100, 1:200, 1:400 e 1:800. Os LD e LQ do ELISA para IC-LDL(-) foram 0,023 g/L e 0,034 g/L de IC-LDL(-), respectivamente. Os plasmas apresentaram linearidade quando diluídos 1:12,5, 1:25, 1:50 e 1:100. Os três ELISAs apresentaram precisão intra e inter-ensaios e recuperação dentro dos limites requeridos para imunoensaios. Para o desenvolvimento de um biossensor para LDL(-), uma proteína recombinante, denominada GFP5-scFv, foi expressa em bactérias Escherichia coli da linhagem BL21(DE3). Para obtenção dessa proteína foi realizada a inserção da sequência de DNA de um fragmento variável de cadeia única (scFv) anti-LDL(-) em um vetor bacteriano com a sequência de DNA da proteína verde fluorescente (GFP5). Dessa forma, a GFP5-scFv é fluorescente e tem afinidade pela LDL(-)...

The electronegative low-density lipoprotein (LDL) is an important antigen involved in the pathogenesis of atherosclerosis. The LDL(-) causes inflammatory and immune response, leading to production of autoantibodies anti-LDL(-) and the subsequent formation of immune complexes (IC-LDL), which also contribute to the atherosclerotic process. Thus, this study aimed to develop and validate immunoassays for quantification of plasma LDL(-), anti-LDL(-) and LDL-IC(-) as well as developing tools applicable to a biosensor for LDL(-). After the standardization of each ELISA, were evaluated the performance characteristics of methods: limits of detection (LD) and quantification (LQ), intra- and inter-assays precision, accuracy, linearity of dilution and interferences. The LD and LQ of LDL(-) ELISA were 0.423 mg/L and 0.517 mg/L of LDL(-), respectively. Plasmas showed linearity when diluted 1:1000, 1:2000, 1:4000 and 1:8000. The LD and LQ of anti-LDL(-) ELISA were 0.0028 mg/L and 0.0032 mg/L of anti-LDL(-), respectively. Plasmas showed linearity when diluted 1:100, 1:200, 1:400 and 1:800. The LD and LQ of IC-LDL(-) ELISA were 0.023 g/L and 0.034 g/L of LDL-IC(-), respectively. Plasma showed linearity when diluted 1:12.5, 1:25, 1:50 and 1:100. The three ELISAs showed good intra- and inter-assays precision and recovery within the limits required for immunoassays. To develop a biosensor for LDL(-), a recombinant protein, called GFP5-scFv was expressed in Escherichia coli BL21(DE3) strain. The obtainment of this protein was performed inserting the DNA sequence of an anti-LDL(-) single chain variable fragment (scFv) in a bacterial vector with a DNA sequence of green fluorescent protein (GFP5). Thus, the scFv-GFP5 is fluorescent and has affinity for LDL(-). Were also synthesized gold nanoparticles, which can be efficiently used for quenching the fluorescence emission of GFP5-scFv. Therefore, immunoassays validated and developed applications for the biosensor are tools that have...

Current Status of External Quality Assessment of Fecal Occult Blood Test / 대한진단검사의학회지

BACKGROUND: Nationwide external quality assessment (EQA) of the fecal occult blood test (FOBT) in Korea was first introduced in 2007-2009. The EQA results were analyzed to assess the current status of FOBT and to plan the continuation of the EQA program. METHODS: The surveys included 40 hospitals in the preliminary survey conducted in 2007, 249 general hospitals in 2008, and 389 hospitals in 2009. In the surveys, the participating hospitals provided the results of the distributed materials and replies to the questionnaire on the FOBT test procedures and quality controls. RESULTS: In the surveys conducted between 2007 and 2009, a total of 650 institutes submitted 653 test system results; 3 institutes used 2 kinds of methods. All of the institutes used immunologic methods; 107 institutes (16.5%) used quantitative equipments and 546 institutes (84.0%) used qualitative kits. Most quantitative tests yielded consistent positive or negative results; however, their cut-off and measured values differed according to the equipments used. A low-level material tested in 2007 was negative in the quantitative methods but positive in some qualitative methods because of lower detection limits. The discordance rates among quantitative tests were 3.2% in 2007, 4.4% in 2008, and 0% in 2009 and the rates among qualitative tests were 13.8% in 2008 and 2.6% in 2009. Semi-solid EQA materials showed the ability to evaluate the overall test procedures with acceptable stability. CONCLUSIONS: In the first Korean FOBT EQA, commercially available EQA materials were proven to be stable. Continuation of the EQA program and further education of laboratory personnel are needed to reduce inconsistency in results. Further, the test kit, procedures, and result reports must be standardized.

Linfoma de Hodgkin: relato de caso / Hodgkin lymphoma: case report

JUSTIFICATIVA E OBJETIVOS: O presente estudo relata o caso de apresentação indolente e pouco usual do linfoma de Hodgkin, uma vez que o envolvimento primário de sistema nervoso central (SNC) ocorre em menos de 1% da doença em estadio avançado. RELATO DO CASO: Paciente do sexo feminino, 24 anos, cuja investigação diagnóstica foi realizada no Hospital São Paulo (HSP), vinculado à Universidade Federal de São Paulo (UNIFESP), durante o ano de 2007. Exames laboratoriais e de imagem, além de biópsia linfonodal associada à imuno-histoquímica permitiram a elucidação do correto diagnóstico e posterior encaminhamento para tratamento especializado, em outra instituição. CONCLUSÃO: Destarte, ressalta-se a importância de uma avaliação clínica completa e abrangente, devendo fazer parte do diagnóstico diferencial expressões atípicas de certas doenças.(AU)

BACKGROUND AND OBJECTIVES: This study reports an indolent and unusual presentation of Hodgkin Disease, since involvement of central nervous system is rare, occurring less than 1% in advanced stages. CASE REPORT: Female patient, a 24-years-old, whose diagnostic investigation took place in São Paulo Hospital (HSP), bounded to Federal University of São Paulo (UNIFESP), in 2007. Laboratorial and image exams, associated with lymph node biopsy and immunohistochemistry allowed correct diagnosis explanation, and subsequent guiding for specific therapy, in another institution. CONCLUSION: Thus, the case points out the importance of a complete and wide clinic evaluation, including atypical expressions of certain diseases in the differential diagnosis.(AU)