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1.
Appl Microbiol Biotechnol ; 104(24): 10601-10612, 2020 Dec.
Article in English | MEDLINE | ID: mdl-33141297

ABSTRACT

This study evaluated the reliability of MALDI-TOF MS coupled with statistical tools for the identification of Streptococcus mutans in comparison with PCR-based techniques. Bacterial isolates were identified and serotyped by conventional PCR, using S. mutans species and serotype-specific primers. For bacterial identification, mass spectra data from S. mutans and other streptococci were compared with Biotyper V 3.1 database and the mass peak lists were examined by cluster and principal component (PCA) analysis. Identification of potential biomarkers was performed using UniProtKB/Swiss-Prot and UniProtKB/TrEMBL databases and BLAST tool of the NCBI database. PCR identified 100% of the isolates as S. mutans. S. mutans strains were typed as serotypes c (85.6%), e (8.6%), k (4.8%), and f (0.9%). Although only the 70% of the strains tested were identified at species level by the Biotyper database, PCA and cluster analysis of mass peaks allowed the identification of 100% S. mutans isolates and its differentiation from the other oral and non-oral streptococci. One mass peak at m/z value of 9572.73 was identified as species-specific biomarker for S. mutans. No biomarkers were identified for S. mutans serotypes. KEY POINTS: • MALDI-TOF MS coupled with statistical tools for the identification of S. mutans. • Detection of species identifying biomarkers by MALDI-TOF MS. • PCR identification and serotyping of S. mutans from saliva samples.


Subject(s)
Streptococcus mutans , Streptococcus , Lasers , Reproducibility of Results , Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization , Streptococcus mutans/genetics
2.
Int Microbiol ; 23(4): 589-596, 2020 Nov.
Article in English | MEDLINE | ID: mdl-32445163

ABSTRACT

The aim of this study was to determine the prevalence of Streptococcus mutans and its serotypes in samples from oral cavity of young Galician population and their relationship with the oral health state. The variables generally associated with dental caries, such as salivary flow rate, buffering capacity, eating habits, and lifestyle, were also analysed. No relationship was found between the variables studied and the presence of S. mutans in the oral cavity or the existence of dental caries. Presumptive strains of S. mutans were isolated from saliva samples from 48% of the analysed population. The use of conventional microbiological methods, API 20 Strep system, and species-specific polymerase chain reaction (PCR) allowed to substantiate the identity of the strains as S. mutans. Multiplex PCR protocols, developed in this study for the simultaneous detection of S. mutans and serotypes c, e, and f and for detection of S. mutans and serotype k, also confirmed this result and demonstrated that serotype c was predominant in the studied young Galician population (86%). Serotypes e (8%), k (3%), and f (2%) were also detected. Serotype c was detected in carious and caries-free subjects, while the remaining serotypes were only found in subjects with caries.


Subject(s)
Dental Caries/microbiology , Molecular Typing/methods , Mouth/microbiology , Serotyping/methods , Streptococcus mutans/classification , Streptococcus mutans/isolation & purification , Feeding Behavior , Female , Humans , Male , Oral Health , Polymerase Chain Reaction , Saliva/microbiology , Saliva/physiology , Spain , Streptococcus mutans/genetics
3.
Enferm Infecc Microbiol Clin (Engl Ed) ; 37(4): 231-238, 2019 Apr.
Article in English, Spanish | MEDLINE | ID: mdl-30929668

ABSTRACT

BACKGROUND: Reflex testing of antibodies and viral load in the same sample for diagnosing hepatitis C virus infection speeds up access to treatment. However, how hepatitis C is diagnosed in Spanish hospitals is unknown. OBJECTIVE: To describe the available resources and procedures for the diagnosis of hepatitis C virus infection in Spain. METHODS: Survey sent to public and private Spanish hospitals with teaching accreditation with at least 200 beds. RESULTS: Of the 160 hospitals that met the inclusion criteria, 90 centres (response rate 56.3%) completed the survey. Two hospitals (2.2%) have no diagnostic resources, 15 (16.7%) can only test for anti-hepatitis C virus(Ab), 9 (10.0%) for Ab and viral load, 47 (52.2%) for Ab, viral load and genotype, 2 (2.2%) for Ab, viral load and core antigen, and 15 (16.7%) can perform Ab, core antigen, viral load and genotype tests. When an Ab test is positive, 28 (31.1%) hospitals perform reflex testing. When an active infection is diagnosed, some communication strategy is used in 62 (68.9%) hospitals. Approximately 44.2% of the respondents believe that all determinations needed to reach a definitive diagnosis should be done on a single blood sample. CONCLUSION: Although 81% of Spanish hospitals have the resources to perform reflex hepatitis C virus infection testing, it is only done in 31%, and less than a half of respondents believe that the definitive diagnosis should be performed on a single sample.


Subject(s)
Hepatitis C, Chronic/diagnosis , Cross-Sectional Studies , Health Care Surveys , Hepatitis C, Chronic/virology , Humans , Spain , Virology/methods
4.
Enferm. infecc. microbiol. clín. (Ed. impr.) ; 37(4): 231-238, abr. 2019. tab
Article in Spanish | IBECS | ID: ibc-189202

ABSTRACT

ANTECEDENTES: El diagnóstico de la infección por el virus de la hepatitis C en un solo paso (anticuerpos y carga viral en una misma muestra) disminuiría el tiempo de acceso al tratamiento. Se desconoce el proceso del diagnóstico de la hepatitis C en los hospitales españoles. OBJETIVO: Describir los recursos diagnósticos disponibles y el proceso del diagnóstico de la infección por virus de la hepatitis C en España. MÉTODOS: Encuesta dirigida a hospitales españoles, públicos o privados con acreditación docente, de 200 camas o más. RESULTADOS: De los 160 hospitales con criterios de inclusión, respondieron 90 (tasa de respuesta del 56,3%). Dos centros (2,2%) no disponen de ningún recurso diagnóstico, 15 (16,7%) solo pueden realizar determinación de anticuerpos (Ac), 9 (10%) de Ac y carga viral (CV), 47 (52,2%) de Ac, CV y genotipo, 2 (2,2%) de Ac, CV y antígeno core, y 15 (16,7%) de Ac, antígeno core, CV y genotipo. Ante un resultado de Ac positivo, 28 (31,1%) hospitales hacen diagnóstico virológico en la misma muestra. Cuando hay una infección activa, se utiliza alguna estrategia de comunicación en 62 (68,9%) hospitales. El 44,2% de los respondedores cree que las determinaciones para llegar a un diagnóstico definitivo se deben hacer con una muestra única de sangre. CONCLUSIONES: Aunque el 81% de los hospitales dispone de medios para realizar el diagnóstico de la infección por virus de la hepatitis C en un solo paso, únicamente lo hace el 31%. Menos de la mitad de los encuestados cree que el diagnóstico definitivo se debe hacer con una única muestra


BACKGROUND: Reflex testing of antibodies and viral load in the same sample for diagnosing hepatitis C virus infection speeds up access to treatment. However, how hepatitis C is diagnosed in Spanish hospitals is unknown. OBJECTIVE: To describe the available resources and procedures for the diagnosis of hepatitis C virus infection in Spain. METHODS: Survey sent to public and private Spanish hospitals with teaching accreditation with at least 200 beds. RESULTS: Of the 160 hospitals that met the inclusion criteria, 90 centres (response rate 56.3%) completed the survey. Two hospitals (2.2%) have no diagnostic resources, 15 (16.7%) can only test for anti-hepatitis C virus(Ab), 9 (10.0%) for Ab and viral load, 47 (52.2%) for Ab, viral load and genotype, 2 (2.2%) for Ab, viral load and core antigen, and 15 (16.7%) can perform Ab, core antigen, viral load and genotype tests. When an Ab test is positive, 28 (31.1%) hospitals perform reflex testing. When an active infection is diagnosed, some communication strategy is used in 62 (68.9%) hospitals. Approximately 44.2% of the respondents believe that all determinations needed to reach a definitive diagnosis should be done on a single blood sample. CONCLUSION: Although 81% of Spanish hospitals have the resources to perform reflex hepatitis C virus infection testing, it is only done in 31%, and less than a half of respondents believe that the definitive diagnosis should be performed on a single sample


Subject(s)
Humans , Hepatitis C, Chronic/diagnosis , Hepatitis C, Chronic/virology , Cross-Sectional Studies , Spain , Virology/methods , Health Care Surveys
5.
Enferm. infecc. microbiol. clín. (Ed. impr.) ; 37(4): 256-259, abr. 2019. tab
Article in Spanish | IBECS | ID: ibc-189206

ABSTRACT

INTRODUCCIÓN: Presentamos el mayor estudio realizado en Galicia sobre prevalencia y distribución de genotipos/subtipos del VHC. MÉTODOS: Estudio retrospectivo recopilando al total de pacientes crónicamente infectados por VHC entre el 1 de enero del 2000 y el 31 de diciembre del 2015 en 3 de las principales áreas sanitarias: Santiago, Pontevedra y Vigo. Recogimos datos sobre prevalencia y distribución de genotipos/subtipos, vías de transmisión, género, edad y coinfección con VIH/VHB. RESULTADOS: Reclutamos a 4.469 pacientes, con una mediana de edad 50 años (RIC 57-45), el 72,3% hombres, el 0,4% coinfectado con otro genotipo, el 20,6% coinfectado con VIH y un 35,2% con VHB. Transmisión parenteral (83,1%), desconocida (15,3%), sexual (1,4%) y vertical (0,2%). Distribución de genotipos: 62,9% VHC-1 (29,2% VHC-1a y 31,9% VHC-1b), 3,4% VHC-2, 21,0% VHC-3, 12,6% VHC-4 y 0,1% VHC-5. CONCLUSIÓN: La distribución de genotipos muestra diferencias significativas respecto a la observada en España. Dicha distribución varía con la edad, el género, la coinfección con VIH o VHB, y dentro de áreas geográficas


INTRODUCTION: We present the largest study conducted in Galicia on the prevalence and distribution of HCV genotypes/subtypes. METHODS: Retrospective study collecting the total number of patients chronically infected by HCV between 2000.01.01 to 2015.12.31 in 3 of the main health areas: Santiago, Pontevedra and Vigo. RESULTS: We collected a total of 4469 patients. The median age was 50 years (IQR 57-45), 72,3% were men, 0,4% were coinfected with another genotype, 20,6% were coinfected with HIV and 35.2% with HBV. The main route of transmission was parenteral (83,1%), followed by unknown (15,3%), sexual (1,4%) and vertical (0,2%). The distribution of genotypes was: 62,9% HCV-1 (29,2% HCV-1a and 31,9% HCV-1b), 3,4% HCV-2, 21,0% HCV-3, 12,6% HCV-4 and 0,1% HCV-5. CONCLUSION: The distribution of genotypes in Galicia shows significant differences with respect to that observed in Spain. This distribution varies with age, gender, coinfection with HIV and/or HBV, and within geographical areas


Subject(s)
Humans , Male , Female , Middle Aged , Hepacivirus/genetics , Hepatitis C, Chronic/epidemiology , Hepatitis C, Chronic/virology , Genotype , Hepacivirus/classification , Retrospective Studies , Spain/epidemiology , Time Factors
6.
Rev. esp. enferm. dig ; 111(1): 10-16, ene. 2019. tab, graf
Article in Spanish | IBECS | ID: ibc-182154

ABSTRACT

Antecedentes: la implementación de estrategias de diagnóstico de la hepatitis C en un solo paso contribuiría a disminuir el tiempo de acceso al tratamiento, evitando el progreso de la enfermedad y sus complicaciones y facilitando la eliminación de la enfermedad. Objetivo: evaluar la validez y la seguridad de las posibles estrategias del diagnóstico de la infección por virus de la hepatitis C (VHC) en un solo paso, así como sus costes y eficiencia. Métodos: estudio de evaluación económica de la eficiencia (coste/eficacia) mediante la construcción de árboles de decisión con análisis de sensibilidad determinista. La perspectiva del análisis es la del financiador (Sistema Nacional de Salud), que solo considera los costes directos. Se han considerado únicamente los costes diferenciales (costes de los test analíticos realizados). El ámbito del estudio es España. Se define la eficacia de una estrategia diagnóstica como el porcentaje de pacientes con infección activa por VHC con diagnóstico positivo y la eficiencia, como el coste de diagnosticar correctamente un paciente con infección activa. Resultados: todas las estrategias para el diagnóstico de VHC en un solo paso evaluadas presentan validez y seguridad aceptables como consecuencia de la elevada sensibilidad y especificidad que tienen los test utilizados. La estrategia más eficiente es Ac-Ag, seguida de Ac-Ag-CV y Ac-CV. La estrategia más eficiente es Ac-Ag debido a su menor coste por paciente testado, pero es menos eficaz que Ac-CV. Conclusión: los hallazgos de este estudio pueden contribuir a establecer las estrategias de diagnóstico más adecuadas en un solo paso teniendo en cuenta su eficacia y eficiencia


Background: implementing one-step strategies for hepatitis C diagnosis would help shorten the time to treatment access. Thus avoiding disease progression and complications, while facilitating hepatitis C virus (HCV) elimination. Objective: to assess the validity and certainty of potential one-step strategies for the diagnosis of HCV infection and their associated cost and efficiency. Methods: the study design is an economic appraisal of efficiency (cost/efficacy) using decision trees and deterministic sensitivity analysis. The analysis was performed from the payer perspective (Spanish National Health System), which exclusively considers the direct costs. Only the differential costs (diagnostic testing costs) were taken into account and the study was set in Spain. The efficacy of a diagnostic strategy was defined as the percentage of patients with an active HCV infection who received a positive diagnosis and the efficiency was defined as the cost per patient with a correctly diagnosed and active infection. Results: the one-step strategies evaluated for the diagnosis of HCV had an acceptable validity and certainty due to the high sensitivity and specificity of the considered tests. The Ab-Ag strategy was the most efficient, followed by Ab-Ag-VL and Ab-VL. Ab-Ag was the most efficient due to the lower cost per patient tested, although the efficacy was lower than the Ab-VL efficacy. Conclusion: the study findings may help to establish more appropriate one-step diagnostic approaches whilst considering the efficacy and efficiency


Subject(s)
Humans , Hepatitis C, Chronic/diagnosis , Mass Screening/economics , Health Care Costs/statistics & numerical data , Antiviral Agents/therapeutic use , Hepatitis C, Chronic/drug therapy , Early Diagnosis , National Health Strategies , Point-of-Care Testing/trends , Decision Trees
7.
Rev Esp Enferm Dig ; 111(1): 10-16, 2019 Jan.
Article in English | MEDLINE | ID: mdl-30561219

ABSTRACT

BACKGROUND: implementing one-step strategies for hepatitis C diagnosis would help shorten the time to treatment access. Thus avoiding disease progression and complications, while facilitating hepatitis C virus (HCV) elimination. OBJECTIVE: to assess the validity and certainty of potential one-step strategies for the diagnosis of HCV infection and their associated cost and efficiency. METHODS: the study design is an economic appraisal of efficiency (cost/efficacy) using decision trees and deterministic sensitivity analysis. The analysis was performed from the payer perspective (Spanish National Health System), which exclusively considers the direct costs. Only the differential costs (diagnostic testing costs) were taken into account and the study was set in Spain. The efficacy of a diagnostic strategy was defined as the percentage of patients with an active HCV infection who received a positive diagnosis and the efficiency was defined as the cost per patient with a correctly diagnosed and active infection. RESULTS: the one-step strategies evaluated for the diagnosis of HCV had an acceptable validity and certainty due to the high sensitivity and specificity of the considered tests. The Ab-Ag strategy was the most efficient, followed by Ab-Ag-VL and Ab-VL. Ab-Ag was the most efficient due to the lower cost per patient tested, although the efficacy was lower than the Ab-VL efficacy. CONCLUSION: the study findings may help to establish more appropriate one-step diagnostic approaches whilst considering the efficacy and efficiency.


Subject(s)
Cost-Benefit Analysis , Decision Trees , Hepatitis C/diagnosis , Diagnostic Tests, Routine/economics , Disease Progression , Hepacivirus/immunology , Hepatitis C/economics , Hepatitis C/virology , Hepatitis C Antibodies/analysis , Hepatitis C Antigens/analysis , Humans , Insurance, Health, Reimbursement , National Health Programs/economics , Predictive Value of Tests , Reproducibility of Results , Sensitivity and Specificity , Viral Load
8.
Enferm Infecc Microbiol Clin (Engl Ed) ; 37(4): 256-259, 2019 Apr.
Article in English, Spanish | MEDLINE | ID: mdl-29759421

ABSTRACT

INTRODUCTION: We present the largest study conducted in Galicia on the prevalence and distribution of HCV genotypes/subtypes. METHODS: Retrospective study collecting the total number of patients chronically infected by HCV between 2000.01.01 to 2015.12.31 in 3of the main health areas: Santiago, Pontevedra and Vigo. RESULTS: We collected a total of 4469 patients. The median age was 50 years (IQR 57-45), 72,3% were men, 0,4% were coinfected with another genotype, 20,6% were coinfected with HIV and 35.2% with HBV. The main route of transmission was parenteral (83,1%), followed by unknown (15,3%), sexual (1,4%) and vertical (0,2%). The distribution of genotypes was: 62,9% HCV-1 (29,2% HCV-1a and 31,9% HCV-1b), 3,4% HCV-2, 21,0% HCV-3, 12,6% HCV-4 and 0,1% HCV-5. CONCLUSION: The distribution of genotypes in Galicia shows significant differences with respect to that observed in Spain. This distribution varies with age, gender, coinfection with HIV and/or HBV, and within geographical areas.


Subject(s)
Hepacivirus/genetics , Hepatitis C, Chronic/epidemiology , Hepatitis C, Chronic/virology , Female , Genotype , Hepacivirus/classification , Humans , Male , Middle Aged , Prevalence , Retrospective Studies , Spain/epidemiology , Time Factors
9.
Rev. esp. quimioter ; 30(6): 468-471, dic. 2017. graf
Article in Spanish | IBECS | ID: ibc-169402

ABSTRACT

Introducción. La edad y sexo a la que se produce la primoinfección por VEB en España es un tema poco estudiado. El objetivo de este trabajo es conocer su relación con la presencia de la primoinfección por VEB entre los años 2006 a 2015 en nuestra área sanitaria. Pacientes y métodos. Se estudiaron 578 pacientes del área sanitaria de Santiago de Compostela con patrones serológicos de primoinfección por VEB, resultados serológicos de IgM-VCA positivo, IgG-VCA positivo y EBNA negativo correspondientes a los años 2006 a 2015. Resultados. Se encontraron 260/578 (45%) adolescentes (11-19 años). En número de casos por edad se observaron dos máximos, a los 2 y 16 años. Entre los 14-19 años, un 62% (79/127) de mujeres tenían entre 14-16 años, mediana de edad 15,8 años (IQ: 14,8-16,4) frente a un 48% (49/102) de hombres, mediana de edad 16 años (IQ: 15,7-16,6) (p=0,032, p=0,02, respectivamente). Conclusiones. Como en nuestro estudio, en los países desarrollados la mayoría de primoinfecciones por VEB ocurren en la adolescencia y se observa una distribución bimodal en relación a la edad. Durante la adolescencia las mujeres adquieren antes que los hombres la primoinfección por VEB (AU)


Introduction. In Spain, the age and sex to which the primary infection by EBV is produced is poorly studied. The objective of this work is to know its relation with the presence of the primary infection by EBV between the years 2006 and 2015 in our health area. Patients and methods. From the Santiago de Compostela health area between 2006 and 2015, 578 patients with serological patterns of EBV primoinfection were selected. This patients presented serological results of IgM-VCA positive, IgG-VCA positive and EBNA negative. Results. We found 260/578 (45%) adolescents (11- 19 years). In the number of cases by age the maximum was observed, at 2 and 16 years. Between 14-19 years, 62% (79/127) of women between 14-16 years of age, median age 15.8 years (IQ: 14.8-16.4) compared to 48% (49/102) of men, median age 16 years (IQ: 15.7-16.6) (p = 0.032, p = 0.02, respectively). Conclusions. As in our study, in the developed countries the majority of primary infections by EBV occur in adolescence and a bimodal distribution is observed in relation to age. During adolescence women acquire before men the first infection by EBV (AU)


Subject(s)
Humans , Male , Female , Child , Adolescent , Herpesvirus 4, Human/isolation & purification , Epstein-Barr Virus Infections/epidemiology , Infectious Mononucleosis/epidemiology , Age and Sex Distribution , Retrospective Studies
10.
Enferm. infecc. microbiol. clín. (Ed. impr.) ; 33(8): e44-e52, oct. 2015. ilus, tab
Article in Spanish | IBECS | ID: ibc-143292

ABSTRACT

El presente documento intenta reflejar y actualizar las principales tareas y cometidos que un laboratorio de microbiología debería tener para realizar el diagnóstico y seguimiento de los pacientes infectados por el VIH. Se distribuye en 3 apartados: en el primero se trata el diagnóstico serológico, que en los últimos años ha sufrido una importante renovación, y en el que hemos procurado adecuarnos a las demandas diagnósticas y epidemiológicas actuales, para que desde los laboratorios podamos contribuir a no perder oportunidades de diagnóstico. En una segunda parte se describe la determinación de la carga viral plasmática, y se hace una exhaustiva revisión de los avances tecnológicos y de las recomendaciones actuales, además de abordar un tema de enorme interés clínico, la significación de la viremia persistente de bajo grado. Finalmente, en el tercer apartado se desarrolla el tema de las resistencias a los fármacos antirretrovirales tanto en pacientesnaive como en fracaso, analizando la transcriptasa reversa, la proteasa y la integrasa, e incorporando como novedades las técnicas de determinación del tropismo viral, y el papel de las variantes minoritarias


This document attempts to update the main tasks and roles of the Clinical Microbiology laboratory in HIV diagnosis and monitoring. The document is divided into three parts. The first deals with HIV diagnosis and how serological testing has changed in the last few years, aiming to improve diagnosis and to minimize missed opportunities for diagnosis. Technological improvements for HIV Viral Load are shown in the second part of the document, which also includes a detailed description of the clinical significance of low-level and very low-level viremia. Finally, the third part of the document deals with resistance to antiretroviral drugs, incorporating clinical indications for integrase and tropism testing, as well as the latest knowledge on minority variants


Subject(s)
Humans , AIDS Serodiagnosis/methods , HIV Infections/microbiology , HIV Seropositivity/microbiology , Viral Load/methods , Anti-Retroviral Agents/therapeutic use , Drug Resistance, Viral
12.
Enferm Infecc Microbiol Clin ; 33(8): e44-52, 2015 Oct.
Article in Spanish | MEDLINE | ID: mdl-25444049

ABSTRACT

This document attempts to update the main tasks and roles of the Clinical Microbiology laboratory in HIV diagnosis and monitoring. The document is divided into three parts. The first deals with HIV diagnosis and how serological testing has changed in the last few years, aiming to improve diagnosis and to minimize missed opportunities for diagnosis. Technological improvements for HIV Viral Load are shown in the second part of the document, which also includes a detailed description of the clinical significance of low-level and very low-level viremia. Finally, the third part of the document deals with resistance to antiretroviral drugs, incorporating clinical indications for integrase and tropism testing, as well as the latest knowledge on minority variants.


Subject(s)
AIDS Serodiagnosis , HIV Infections/diagnosis , Viremia/diagnosis , AIDS Serodiagnosis/methods , AIDS Serodiagnosis/trends , Algorithms , Anti-HIV Agents/therapeutic use , Antiretroviral Therapy, Highly Active , CD4 Lymphocyte Count , Drug Resistance, Viral , Drug Therapy, Combination , Female , Genotyping Techniques , HIV Infections/drug therapy , HIV Infections/prevention & control , HIV Infections/virology , HIV-1/genetics , HIV-1/immunology , HIV-1/isolation & purification , HIV-2/genetics , HIV-2/immunology , HIV-2/isolation & purification , Humans , Immunoassay/methods , Infectious Disease Transmission, Vertical , Male , Pregnancy , Pregnancy Complications, Infectious/drug therapy , Viral Load , Viremia/drug therapy , Viremia/virology
16.
Enferm Infecc Microbiol Clin ; 24(9): 541-5, 2006 Nov.
Article in Spanish | MEDLINE | ID: mdl-17125671

ABSTRACT

OBJECTIVE: The aim of this study is to assess a real-time PCR technique on the LightCycler 2.0 with SYBR-Green I detection as compared to another real-time PCR method based on detection with FRET (fluorescence resonance energy transfer) probes for the quantification of CMV DNA. METHODS: The two real-time PCR methods were used to test plasma samples from immunocompromised patients with clinically suspected CMV disease, patients under follow-up without symptoms, and healthy adults. A standard curve for quantitative analysis by the SYBR-Green I method was performed with 10-fold diluted solutions of DNA from the CMV Towne strain (ATCC VR-977) cultured in MRC-5 monolayer. In addition, frozen samples from patients positive for CMV pp65 antigenemia were also analyzed and results compared using the two real time PCR methods. RESULTS: The real-time PCR technique using SYBR-Green I on the LightCycler 2.0 was a highly specific, fast, simple and reliable test to quantify CMV; moreover, it was cost-effective. CONCLUSION: Quantification of CMV DNA in plasma using this sensitive, fast, low-cost method was advantageous for the diagnosis and follow up of patients with opportunistic CMV infection, which are increasingly more frequent in our daily hospital clinical practice.


Subject(s)
Antigens, Viral/blood , Computer Systems , Cytomegalovirus Infections/blood , DNA, Viral/blood , Fluorescent Dyes/analysis , Opportunistic Infections/blood , Organic Chemicals/analysis , Polymerase Chain Reaction/methods , Viremia/blood , AIDS-Related Opportunistic Infections/blood , AIDS-Related Opportunistic Infections/diagnosis , AIDS-Related Opportunistic Infections/virology , Adult , Benzothiazoles , Bone Marrow Transplantation , Cell Line , Child , Computer Systems/economics , Cytomegalovirus/genetics , Cytomegalovirus/immunology , Cytomegalovirus/isolation & purification , Cytomegalovirus Infections/diagnosis , Cytomegalovirus Infections/virology , Diamines , Fluorescence Resonance Energy Transfer , Humans , Immunocompromised Host , Infant, Newborn , Neoplasms/complications , Neutrophils/virology , Opportunistic Infections/diagnosis , Opportunistic Infections/virology , Phosphoproteins/blood , Polymerase Chain Reaction/economics , Polymerase Chain Reaction/instrumentation , Polymerase Chain Reaction/standards , Postoperative Complications/blood , Postoperative Complications/diagnosis , Postoperative Complications/virology , Quinolines , Reagent Kits, Diagnostic , Reference Standards , Sensitivity and Specificity , Viral Matrix Proteins/blood , Viremia/diagnosis , Viremia/virology
17.
Enferm. infecc. microbiol. clín. (Ed. impr.) ; 24(9): 541-545, nov. 2006. tab, graf
Article in Es | IBECS | ID: ibc-051038

ABSTRACT

Objetivo. El objetivo del presente estudio es evaluar una técnica rápida y sencilla de reacción en cadena de la polimerasa (PCR) en tiempo real en LightCycler 2.0 revelada con SYBR-Green I, comparándola con otra técnica de PCR en tiempo real que utiliza sondas FRET (fluorescence resonance energy transfer) para revelar la amplificación. Métodos. Los dos métodos de PCR en tiempo real se compararon utilizando muestras de plasma de pacientes inmunodeprimidos con sospecha clínica de enfermedad por citomegalovirus (CMV), de pacientes monitorizados sin sintomatología, y de adultos sanos. El estudio se completó con otras muestras de plasma congeladas de casos positivos por antigenemia pp65, y con ADN de CMV de la cepa Towne (ATCC VR-977) obtenido de cultivo en MRC-5, con el que elaboramos una curva estándar para su cuantificación. Resultados. La PCR revelada con SYBR-Green I resultó ser claramente la más rentable por su alta sensibilidad, rapidez y sencilla realización, además de su bajo coste. Conclusión. La determinación cuantitativa de ADN de CMV en plasma utilizando un método sensible, rápido y de bajo coste, como el que proponemos, supone una clara ventaja para el diagnóstico y seguimiento de estos cuadros, especialmente en hospitales como el nuestro, donde en los últimos años se ha incrementado sensiblemente el número de pacientes susceptibles de padecer esta infección oportunista (AU)


Objective. The aim of this study is to assess a real-time PCR technique on the LightCycler 2.0 with SYBR-Green I detection as compared to another real-time PCR method based on detection with FRET (fluorescence resonance energy transfer) probes for the quantification of CMV DNA. Methods. The two real-time PCR methods were used to test plasma samples from immunocompromised patients with clinically suspected CMV disease, patients under follow-up without symptoms, and healthy adults. A standard curve for quantitative analysis by the SYBR-Green I method was performed with 10-fold diluted solutions of DNA from the CMV Towne strain (ATCC VR-977) cultured in MRC-5 monolayer. In addition, frozen samples from patients positive for CMV pp65 antigenemia were also analyzed and results compared using the two real time PCR methods. Results. The real-time PCR technique using SYBR-Green I on the LightCycler 2.0 was a highly specific, fast, simple and reliable test to quantify CMV; moreover, it was cost-effective. Conclusion. Quantification of CMV DNA in plasma using this sensitive, fast, low-cost method was advantageous for the diagnosis and follow up of patients with opportunistic CMV infection, which are increasingly more frequent in our daily hospital clinical practice (AU)


Subject(s)
Humans , Polymerase Chain Reaction/methods , Cytomegalovirus/genetics , Computer Systems , DNA, Viral/analysis , Cytomegalovirus Infections/blood , Nucleic Acid Amplification Techniques/methods
18.
Enferm Infecc Microbiol Clin ; 24(4): 264-76, 2006 Apr.
Article in Spanish | MEDLINE | ID: mdl-16725087

ABSTRACT

Viral hepatitis is an infectious disease affecting the liver. At this time, five different human hepatitis viruses are recognized and characterized in detail: Hepatitis A virus (HAV), hepatitis B virus (HBV), hepatitis C virus (HCV), hepatitis D virus (HDV), and hepatitis E virus (HEV). The hepatitis viruses differ widely in their modes of transmission and clinical features. Whereas all of them can cause acute hepatitis, only HBV, HDV, and HCV cause chronic hepatitis. This article reviews the epidemiology and clinical manifestations of the different types of viral hepatitis.


Subject(s)
Hepatitis, Viral, Human/epidemiology , Adolescent , Adult , Aged , Child , Child, Preschool , Female , Global Health , Hepatitis Viruses/classification , Hepatitis Viruses/isolation & purification , Hepatitis Viruses/physiology , Hepatitis, Viral, Human/classification , Hepatitis, Viral, Human/diagnosis , Hepatitis, Viral, Human/immunology , Hepatitis, Viral, Human/prevention & control , Hepatitis, Viral, Human/transmission , Hepatitis, Viral, Human/virology , Humans , Infant , Male , Middle Aged , Viral Hepatitis Vaccines
19.
Enferm. infecc. microbiol. clín. (Ed. impr.) ; 24(4): 264-276, abr. 2006. tab, graf
Article in Es | IBECS | ID: ibc-047907

ABSTRACT

La hepatitis viral es una enfermedad infecciosa que afecta al hígado. Hasta el momento, han sido reconocidos y caracterizados minuciosamente en humanos cinco diferentes virus de la hepatitis: el virus de la hepatitis A (VHA), el virus de la hepatitis B (VHB), el virus de la hepatitis C (VHC), el virus de la hepatitis D (VHD) y el virus de la hepatitis E (VHE). Los virus de la hepatitis difieren ampliamente en sus formas de transmisión y características clínicas; ya que mientras todos pueden causar hepatitis aguda, sólo el VHB, el VHD y el VHC ocasionan hepatitis crónica. Este artículo presenta una revisión sobre la epidemiología y manifestaciones clínicas de las diferentes hepatitis virales (AU)


Viral hepatitis is an infectious disease affecting the liver. At this time, five different human hepatitis viruses are recognized and characterized in detail: Hepatitis A virus (HAV), hepatitis B virus (HBV), hepatitis C virus (HCV), hepatitis D virus (HDV), and hepatitis E virus (HEV). The hepatitis viruses differ widely in their modes of transmission and clinical features. Whereas all of them can cause acute hepatitis, only HBV, HDV, and HCV cause chronic hepatitis. This article reviews the epidemiology and clinical manifestations of the different types of viral hepatitis (AU)


Subject(s)
Child , Adult , Aged , Child, Preschool , Adolescent , Middle Aged , Humans , Hepatitis, Viral, Human/epidemiology , Viral Hepatitis Vaccines , Global Health , Hepatitis Viruses/classification , Hepatitis Viruses/isolation & purification , Hepatitis Viruses/physiology , Hepatitis, Viral, Human/classification , Hepatitis, Viral, Human/diagnosis , Hepatitis, Viral, Human/immunology , Hepatitis, Viral, Human/prevention & control , Hepatitis, Viral, Human/transmission , Hepatitis, Viral, Human/virology
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