Pruebas moleculares para la detección de SARS-CoV-2 / Molecular tests for the detection of SARS-CoV-2

RESUMEN Introducción: Los coronavirus generan enfermedades respiratorias, hepáticas o neurológicas, con gravedad variable en su huésped. La reacción en cadena de la polimerasa con transcriptasa inversa en tiempo real es la metodología de referencia para confirmar la detección del virus SARS-CoV-2. Se realizó una revisión bibliográfica en el periodo de julio a noviembre de 2021, en los recursos disponibles en MEDLINE, SciELO, Pubmed y Elsevier. Del total de consultas se citaron 35 referencias. Objetivo: Describir las pruebas moleculares para la detección de SARS-CoV-2. Desarrollo: Se han diseñado tres tipos de ensayos de rRT-PCR, para la detección de SARS-CoV-2. Las pruebas descritas son específicas para genes ARN (polymerase dependent RNA - RdRp), genes E (upE) y genes Nnes (N). Los Centros para la Prevención y Control de Enfermedades (CDC) de los EE.UU., han desarrollado la rRT-PCR dirigidos a la proteína N de la nucleocápside del material genético SARS-CoV-2, los ensayos genes E (upE) y gen de la proteína 1b (ORF 1b) se pueden complementar para la detección y confirmación. Conclusiones: Las dianas que se emplean para la detección específica del genoma del SARS-CoV-2 por RT-PCR en tiempo real, se encuentran en las regiones ORF1a y 1b, RdRp, N, S y E del ARN viral.

ABSTRACT Introduction: Coronaviruses cause respiratory, hepatic or neurological diseases, with variable severity in their host. Real-time reverse transcriptase polymerase chain reaction is the reference methodology to confirm the detection of the SARS-CoV-2 virus. A bibliographic review was carried out in the period from July to November 2021, in the resources available in MEDLINE, SciELO, Pubmed and Elsevier. Of the total queries, 35 references were cited. Objective: Describe molecular tests for the detection of SARS-CoV-2. Development: Three types of rRT-PCR assays have been designed for the detection of SARS-CoV-2. The tests described are specific for RNA polymerase dependent RNA (RdRp) genes, E (upE) genes, and Nnes (N) genes. The United States Centers for Disease Control and Prevention (CDC) have developed rRT-PCR directed at the N protein of the nucleocapsid of the SARS-CoV-2 genetic material, the gene E assays (upE) and protein gene 1b (ORF 1b) can be complemented for detection and confirmation. Conclusions: The targets used for the specific detection of the SARS-CoV-2 genome by real-time RT-PCR are found in the ORF1a and 1b, RdRp, N, S and E regions of the viral RNA.

Relación entre los anticuerpos contra antígenos extraíbles del núcleo y las enfermedades del tejido conectivo identificados por Immunoblots en un hospital y universidad de Lima / Relationship between antibodies against extractable nucleus antigens and connective tissue diseases identified by Immunoblot in a hospital in Lima, Peru

Objetivos: Determinar la relación de los anticuerpos con los antígenos del núcleo extraíble y las enfermedades del tejido conectivo identificadas por Immunoblot en un hospital de Lima, Perú. Material y métodos: Estudio de tipo observacional, ciencias básicas, analíticas y transversales, realizado en el Servicio de Inmunología del Hospital Nacional Arzobispo Loayza entre enero de 2018 y junio de 2018. Analizamos 291 historias clínicas de pacientes con enfermedad del tejido conectivo y para la detección de anticuerpos contra los antígenos extraíbles del núcleo se empleó el método de Immunoblots. Resultados: La frecuencia de los anticuerpos contra antígenos nucleares extraíbles en pacientes con enfermedad del tejido conectivo identificados por Immunoblot fue 789 (100%). Se demostró que existe una relación significativa p <0.05 de Anti-histonas (X2 = 64.19; p = 0,000), anti-nucleosomas (X2 = 71,16; p = 0,000), anti-dsDNA (X2 = 71,44; p = 0,000), anti-SM (X2 = 10,08; p = 0,003) y lupus eritematoso sistémico con prueba de Chi-cuadrado de Pearson. Se demostró que existe una relación significativa p <0.05 del Anti-SSA (X2 = 61,33; p = 0.001), anti-SSB (x2 = 51,00; p = 0.001), anti-Ro 52 (X2 = 62,60; p = 0,000) y síndrome de Sjogren con prueba de Chi-cuadrado de Pearson. Se demostró que existe una relación significativa p <0.05 de Anti-CENP B (p = 0.001) y calcinosis, fenómeno de Raynaud, dismotilidad esofágica, esclerodactilia y Telangiectasia (CREST) con Fisher. Conclusiones: Existe relación de anticuerpos con antígenos de núcleo extraíbles y lupus eritematoso sistémico, síndrome de Sjogren, enfermedad mixta del tejido conectivo, enfermedad del CREST, esclerodermia y polimiositis.

Objectives: To determine the relationship of antibodies to extractable nucleus antigens and connective tissue diseases identified by Immunoblot in a hospital in Lima, Peru. Material and methods: Study of the observational type, basic sciences, analytical and transversal, carried out in the Immunology service of the national Hospital Archbishop Loayza between January 2018 and June 2018. We analyzed 291 clinical histories of patients with connective tissue disease and for the detection of antibodies to the extractable antigens of the nucleus the method of Immunoblot was employed. Results: The frequency of the antibodies against extractable nuclear antigens in patients with connective tissue disease identified by Immunoblot was 789 (100%). It was demonstrated that there is significant relationship p < 0.05 of Anti-histones (X2 = 64.19; p = 0,000), anti-nucleosomas (X2 = 71,16; p = 0,000), anti-dsDNA (X2 = 71,44; p = 0,000), anti-SM (X2 = 10,08; p = 0,003) and Lupus Systemic erythematosus with Pearson Chi-square test. It was demonstrated that there is significant relationship p < 0.05 of the Anti-SSA (X2 = 61,33; p = 0.001), anti-SSB (X2 = 51,00; p = 0.001), anti-Ro 52 (X2 = 62,60; p = 0,000) and Sjogren's syndrome with Pearson Chi-square test. It was demonstrated that there is significant relationship p < 0.05 of Anti-CENP B (p = 0.001) and calcinosis, Raynaud's phenomenon, esophageal dysmotility, sclerodactyly and Telangiectasia (CREST) with exact Fisher statistician. Conclusions: There is a relationship of antibodies to extractable nucleus antigens and systemic lupus erythematosus, Sjogren's syndrome, mixed connective tissue disease, calcinosis, Raynaud's phenomenon, esophageal dysmotility, sclerodactyly and Telangiectasias (CREST), Scleroderma and Polymyositis.

Anticuerpos antimitocondriales y anticuerpos contra antígenos extraíbles del núcleo en pacientes con cirrosis biliar primaria / Antimitochondrial Antibodies and antibodies against extractable nucleus antigens in patients with primary biliary cirrhosis

Resumen Objetivo: Determinar la frecuencia de anticuerpos antimitocondriales y de anticuerpos contra antígenos extraíbles del núcleo en pacientes con cirrosis biliar primaria. Material y métodos: Estudio de tipo cuantitativo, observacional y transversal, realizado en el Servicio de Inmunología del Hospital Nacional Arzobispo Loayza entre enero 2018 y marzo 2019. Se revisaron las historias clínicas de 30 pacientes con características presuntivas de cirrosis biliar primaria; para la detección de los anticuerpos antinucleares y anticuerpos antimitocondriales se empleó el kit inmunológico en sangre y observación con microscopio de inmunofluorescencia a 40X y para la detección de los anticuerpos contra antígenos extraíbles del núcleo se empleó el método Immunoblot. Resultados: Se estudiaron 30 pacientes con cirrosis biliar primaria, 20 fueron de sexo femenino (66,7%). El patrón de tinción más frecuente fue el citoplasmático moteado reticular en 17(56,7%), seguido del patrón citoplasmático moteado reticular y patrón moteado en 7(23,3%) pacientes, y en menor frecuencia el patrón citoplasmático moteado reticular y patrón centromérico. Nueve (42,9%) pacientes con cirrosis biliar primaria tenían anti-M2. Se demostró mayor frecuencia, 21(70%) de los pacientes con cirrosis biliar primaria tenían anticuerpos antimitocondriales. Conclusiones: Se encontró alta frecuencia de patrón citoplasmático moteado reticular en pacientes con cirrosis biliar primaria, se demostró asociación significativa con los anti-M2 y anticuerpos antimitocondriales.

Summary Objective: To determine the frequency of antimitochondrial antibodies and antibodies against extractable nucleus antigens in patients with primary biliary cirrhosis. Methods : A quantitative, observational and cross-sectional study was carried out at the Immunology Service of the Arzobispo Loayza National Hospital between January 2018 and March 2019. The medical records of 30 patients with presumptive characteristics of primary biliary cirrhosis were reviewed; for the detection of the antinuclear antibodies and antimitochondrial antibodies, the immunological kit was used in blood and observation with a 40X immunofluorescence microscope, and the Immunoblot method was used for the detection of the antibodies against extractable nucleus antigens. Results: Thirty patients with primary biliary cirrhosis disease were studied, 20 were female (66.7%). The most frequent staining pattern was the reticular mottled cytoplasmic in 17 (56.7%), followed by the reticular mottled cytoplasmic pattern and mottled pattern in 7 (23.3%) patients, and less frequently the reticular mottled cytoplasmic pattern and centromeric. Nine (42.9%) patients with primary biliary cirrhosis had anti-M2. In the present investigation, a higher frequency was demonstrated, 21 (70%) of the patients with primary biliary cirrhosis had antimitochondrial antibodies. Conclusions: A high frequency of reticular mottled cytoplasmic pattern was found in patients with primary biliary cirrhosis; a significant association with anti-M2 and antimitochondrial antibodies was demonstrated.

Staining of antinuclear antibodies and antibodies against removable nuclear antigens in connective tissue diseases

Introduction and objectives: Connective tissue diseases are inflammatory, autoimmune diseases and threaten quality of life. To determine the relationship between staining patterns of antinuclear antibodies and antibodies against extractable nuclear antigens in patients with connective tissue disease. Materials and methods: Observational, basic, analytical and transversal study. Study conducted in the Immunology Service of the Arzobispo Loayza National Hospital between January 2017 and June 2017. We analyzed 291 samples of patients with CTD and for the detection of anti-nuclear antibody staining patterns, the immunological kit and observation with microscope of at 40X Immunofluorescence and for the detection of the antibodies against extractable nuclear antigens. The Immunoblot method was employed. Statistical analyses were carried out with the statistical package SPSS version 21 for Windows. We used the Pearson Chi-square test for the categorical variables, a value of p < 0.05 was considered significant. Results: There was a significant relationship p < 0.05 of the homogeneous pattern, the mottled pattern with Anti-histones (p = 0.000), Anti-nucleosomes (p = 0.000), Anti-Ro 52 (p = 0.000), Anti-SSA (p = 0.001), Anti-SSB (p = 0.003), Anti-dsDNA (p = 0.000) with the Pearson Chi-square test. There was a significant relationship of p < 0.05 of the centromeric pattern with Anti-Cenp B (p = 0.000) with Fisher's exact statistic. Conclusions: There was a significant relationship between the anti-nuclear antibody staining patterns and the antibodies to the core extractable antigens in patients with systemic lupus erythematosus, Sjögren's syndrome, Calcinosis, Raynaud's phenomenon, esophageal Dysmotility, sclerodactyly and Telangiectasia (CREST), Scleroderma and Polymyositis

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Staining of antinuclear antibodies and antibodies against removable nuclear antigens in connective tissue diseases.

INTRODUCTION AND OBJECTIVES: Connective tissue diseases are inflammatory, autoimmune diseases and threaten quality of life. To determine the relationship between staining patterns of antinuclear antibodies and antibodies against extractable nuclear antigens in patients with connective tissue disease. MATERIALS AND METHODS: Observational, basic, analytical and transversal study. Study conducted in the Immunology Service of the Arzobispo Loayza National Hospital between January 2017 and June 2017. We analyzed 291 samples of patients with CTD and for the detection of anti-nuclear antibody staining patterns, the immunological kit and observation with microscope of at 40X Immunofluorescence and for the detection of the antibodies against extractable nuclear antigens. The Immunoblot method was employed. Statistical analyses were carried out with the statistical package SPSS version 21 for Windows. We used the Pearson Chi-square test for the categorical variables, a value of p<0.05 was considered significant. RESULTS: There was a significant relationship p<0.05 of the homogeneous pattern, the mottled pattern with Anti-histones (p=0.000), Anti-nucleosomes (p=0.000), Anti-Ro 52 (p=0.000), Anti-SSA (p=0.001), Anti-SSB (p=0.003), Anti-dsDNA (p=0.000) with the Pearson Chi-square test. There was a significant relationship of p<0.05 of the centromeric pattern with Anti-Cenp B (p=0.000) with Fisherâ¿¿s exact statistic. CONCLUSIONS: There was a significant relationship between the anti-nuclear antibody staining patterns and the antibodies to the core extractable antigens in patients with systemic lupus erythematosus, Sjögrenâ¿¿s syndrome, Calcinosis, Raynaudâ¿¿s phenomenon, esophageal Dysmotility, sclerodactyly and Telangiectasia (CREST), Scleroderma and Polymyositis.

Patrones de tinción de anticuerpos antinucleares identificados por Inmunofluorescencia indirecta en pacientes con enfermedad del tejido conectivo / Antinuclear antibody staining patterns identified by indirect immunofluorescence in patients with connective tissue disease

Objetivos: Determinar la frecuencia de patrones de tinción de anticuerpos antinucleares identificados por inmunofluorescencia indirecta en pacientes con enfermedad del tejido conectivo en un hospital de nivel III de Lima, Perú. Material y métodos: Estudio de tipo cuantitativo, observacional y transversal, realizado en el Servicio de Inmunología del Hospital Nacional Arzobispo Loayza entre enero 2017 y junio 2017. Se revisaron 291 historias clínicas de pacientes con enfermedad del tejido conectivo y con detección de los patrones de tinción de anticuerpos antinucleares en suero, usando la técnica de Inmunofluorescencia Indirecta. Resultados: La frecuencia de estudios de anticuerpos antinucleares fue 322 (100 %), los patrones de anticuerpos antinucleares fueron PCNA6 (1,86%), patrón homogéneo 109(33,85%), patrón centromérico 34(10,56%), patrón moteado 135(41,93%), patrón citoplasmático 25(7,76%), patrón nucleolar 9(2,80%), patrón NUMA 1- Huso Acromático 3(0,93%) y patrón lisosoma 1(0,31%). La positividad de anticuerpos antinucleares se determinó mayormente en pacientes con lupus eritematoso sistémico 85(29.21%), esclerosis sistémica progresiva limitada (CREST) 34(11,68%), síndrome de Sjögren 77(26,46%), esclerodermia 15(5,15%), enfermedades mixtas del tejido conectivo72 (24,74%) y polimiositis 8 (2,75%). Conclusiones: Se encontró elevada frecuencia del patrón homogéneo y del patrón moteado en pacientes con enfermedad del tejido conectivo. El patrón homogéneo se asoció fuertemente al lupus eritematoso sistémico y el patrón moteado se asoció al síndrome de Sjögren. Por lo tanto, el método ANA-IFI puede reducir el gasto de un trabajo inmunológico detallado con una pérdida mínima en la exactitud diagnóstica de la enfermedad. (AU)

Objectives: To determine the antinuclear antibody staining patterns identified by indirect immunofluorescence in patients with connective tissue disease in hospital level III in Lima, Peru. Methods: Cross-sectional study performed at the Servicio de Inmunología of the Hospital Nacional Arzobispo Loayza between January and June 2017. We reviewed 291 clinical charts of patients with connective tissue disease who had determination of antinuclear antibodies by indirect immunofluorescence. Results: 322 determinations were analyzed; patterns detected were PCNA 6 (1.86%); homogeneous pattern 109(33.85%), centromeric pattern 34(10.56%), speckled pattern 135(41.93%), cytoplasmic pattern 25(7.76%), nucleolar pattern 9(2.80%), NUMA 1- Huso achromatic pattern 3(0.93%) and lysosomal pattern 1(0.31%). The spectrum of diseases in which antinuclear antibodies were looked for were systemic lupus 85(29.21%), limited progressive systemic sclerosis CREST) 34(11.68%), Sjögren´s syndrome 77(26.46%), scleroderma 15(5.15%), mixed connective tissue disorders 72 (24.74%) and polymyositis 8 (2.75%). Conclusions: A high frequency of homogeneous and speckled patterns were observed, the former associated with systemic lupus and the latter with Sjögren´s syndrome. Detection of antinuclear antibodies by indirect immunofluorescence is an accurate diagnostic method. (AU)

Contaminación con bacterias patógenas de estetoscopios del personal médico en un hospital de nivel III en Lima, Perú / Contamination of medical stethoscopes with pathogenic bacteria in a level III hospital in Lima, Peru

Objetivos: Determinar el grado de contaminación bacteriana con bacterias patógenas de los estetoscopios del personal médico en un hospital general de Lima, Perú. Material y métodos: Estudio de tipo observacional,descriptivo y transversal, realizado en el Hospital Nacional Arzobispo Loayza, entre los meses de enero y juniodel 2013. Se estudiaron 124 muestras de estetoscopios del personal médico en las siguientes áreas: UCI 20; neonatología 13; quemados 3; medicina 52; emergencia 36. Se recolectaron las muestras con hisopos humedecidos,en condiciones estériles (En presencia de un mechero de vidrio para alcohol) y luego fueron introducidos en tuboscon preparado de caldo BHI (Infusión cerebro corazón) para ser incubados por 24 horas a 37°C; se cultivó en Agarsangre, Agar MacConkey, Agar manitol y Agar cetrimidepara su posterior determinación de bacterias patógenaspor procedimientos bioquímicos ,luego se identificó la susceptibilidad bacteriana con la técnica de Kirby- Bauer...

Objectives: To determine the degree of contamination with pathogenic bacteria by stethoscopes used for medical personnel in a general hospital in Lima, Peru. Methods: Cross sectional study carried-out at Hospital Nacional Arzobispo Loaya between January and June 2013. A total of 124 samples were evaluated from the following areas: 20 from the ICU, 13 from the neonatology service, 3 from the burn unit; 52 from medicine wards, and 36 from the emergency room. The samples were collected using a moist cotton swab in sterile conditions (using a alcohol bunser burner) and introduced in tubes containing brain-heart infusion to be incubated at 37C for 24 hours. The determination of pathogenic bacteria used MacConkey and manitol agar with specific biochemical methods. The Kirby-Bauer method was used to determine the antibiotic susceptibility pattern...