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3.
Salud bienestar colect ; 4(3): 108-128, sept.-dic. 2020. tab, ilus
Article in Spanish | LILACS | ID: biblio-1282578

ABSTRACT

INTRODUCCIÓN: en el mismo año en que se declara Año Internacional de la Enfermería y Partería, la inesperada aparición del nuevo coronavirus SARS-CoV-2,dio un giro a lo que se tenía planeado dentro de los programas de salud a nivel mundial y deja en evidencia las debilidades de los sistemas sanitarios, donde el continente más afectado por dicho virus fue América, ya que sus esfuerzos por contener la pandemia fueron insuficientes, el tiempo de reacción para establecer protocolos de salud fue tardío y la disponibilidad para dotar al personal de salud de equipos de protección fue mínimo, y aun así el accionar del personal sanitario en especial de enfermería. OBJETIVO: describir la situación de enfermería en América, frente a la pandemia Covid-19. METODOLOGÍA: la investigación se realizó mediante un diseño narrativo, de carácter documental, analítico de enfoque cualitativo y método inductivo; obteniendo la información de fuentes secundarias confiables. RESULTADOS Y CONCLUSIONES: la actual pandemia demuestra la importancia de disponer de profesionales de salud en un número adecuado según las necesidades y cuidados que requiere cada paciente; es por esta razón que se precisa que los países inviertan en mejorar las condiciones de trabajo de los profesionales de enfermería, que incluyan equipos de protección individual, apoyo al trabajo en equipo y educación continua en enfermería, lo cual llevará a importantes logros, evidenciando el profesionalismo de enfermería y su entrega absoluta, al aplicar sus cuatro roles fundamentales con el fin de proteger la salud y mejorar la vida de las personas, a pesar de los evidentes riesgos reales y potenciales a los que se enfrentan a nivel laboral.


INTRODUCTION: in the same year in which the International Year of Nursing and Midwifery is declared, the unexpected appearance of the new SARS-CoV-2 coronavirus, gave a turn to what was planned within health programs worldwide and leaves in evidence the weaknesses of the health systems, where the continent most affected by this virus was America, since their efforts to contain the pandemic were insufficient, the reaction time to establish health protocols was late and the availability to provide staff with The health ofprotective equipment was minimal, and even so, the actions of health personnel, especially nursing personnel. OBJECTIVE: to describe the nursing situation in America, in the face of the Covid-19 pandemic. METHODOLOGY: the research was carried out through a narrative, documentary, analytical design with a qualitative approach and an inductive method; obtaining the information from reliable secondary sources. RESULTS AND CONCLUSIONS: the current pandemic shows the importance of having adequate numbers of health professionals according to the needs and care that each patient requires; It is for this reason that it is necessary for countries to invest in improving the working conditions of nursing professionals, which include individual protection equipment, support for teamwork and continuing education in nursing, which will lead to important achievements, evidencing the Nursing professionalism and its absolute dedication, by applying its four fundamental roles in order to protect health and improve people's lives, despite the obvious real and potential risks they face at the work level.


Subject(s)
Humans , Pneumonia, Viral/nursing , Pneumonia, Viral/virology , Coronavirus Infections , COVID-19/epidemiology , Antigens, Viral/analysis , Pneumonia, Viral/diagnostic imaging , Americas/epidemiology , Radiography, Thoracic , Tomography, X-Ray Computed , Polymerase Chain Reaction , Clinical Laboratory Techniques/methods , Nurse's Role , Ecuador/epidemiology , Asymptomatic Infections/epidemiology , Critical Care Nursing , COVID-19 Testing , SARS-CoV-2 , COVID-19/transmission , COVID-19/diagnostic imaging
4.
Braz. j. infect. dis ; 24(1): 85-88, Feb. 2020. tab, graf
Article in English | LILACS | ID: biblio-1089325

ABSTRACT

ABSTRACT The antigenic potential of seven immunogenic peptides of the dengue virus was evaluated in the sera of patients with dengue confirmed by IgM/IgG serology. Antibodies IgM and IgG against dengue virus peptides were analyzed by ELISA in 31 dengue sero-positive and 20 sero-negative patients. The P5 peptide showed significant IgG immunoreactivity mostly in the sera of patients with dengue without warning signs in comparison with patients with dengue with warning signs, correlating with mild disease. This finding suggests that the low antibody response against P5 epitope could be a risk factor for higher susceptibility to dengue virus infection with warning signs, and that P5 could be a potential antigen for vaccine development.


Subject(s)
Humans , Male , Female , Child , Adolescent , Adult , Middle Aged , Aged , Young Adult , Peptides/immunology , Viral Envelope Proteins/immunology , Dengue Virus/immunology , Dengue Vaccines , Antibodies, Viral/immunology , Epitopes/immunology , Immunoglobulin G/immunology , Immunoglobulin M/immunology , Enzyme-Linked Immunosorbent Assay , Statistics, Nonparametric , Dengue/immunology , Dengue/prevention & control , Antibody Formation , Antigens, Viral/immunology
5.
Chinese Journal of Biotechnology ; (12): 1314-1322, 2020.
Article in Chinese | WPRIM | ID: wpr-826845

ABSTRACT

To screen the best genotypeⅠJapanese encephalitis virus subunit vaccine candidate antigens, the prMEIII gene, the polytope gene and the prMEIII-polytope fusion gene of the GenotypeⅠJapanese encephalitis virus GS strain were cloned into prokaryotic expression vector pET-30a. The recombinant proteins were obtained after the induction and purification. The prepared recombinant proteins were immunized to mice, and the immunogenicity of the subunit vaccine candidate antigens was evaluated through monitoring the humoral immune response by ELISA, detecting the neutralizing antibody titer by plaque reduction neutralization test, and testing the cell-mediated immune response by lymphocyte proliferation assay and cytokine profiling. The recombinant proteins with the molecular weights of 35 (prMEIII), 28 (polytope antigen) and 57 kDa (prMEIII-polytope) induced strong humoral and cellular immune responses in mice. Compared with prMEIII-polytope and polytope proteins, the prMEIII protein induced a significant expression of IL-2 and IFN-γ (P0.05). The study suggests that the prMEIII protein can be used for the development of the Japanese encephalitis virus subunit vaccine.


Subject(s)
Animals , Antibodies, Viral , Blood , Antigens, Viral , Allergy and Immunology , Encephalitis Virus, Japanese , Allergy and Immunology , Encephalitis, Japanese , Allergy and Immunology , Immunogenicity, Vaccine , Mice , Mice, Inbred BALB C , Vaccines, Subunit , Allergy and Immunology , Viral Vaccines , Allergy and Immunology
6.
Chinese Journal of Biotechnology ; (12): 2066-2075, 2020.
Article in Chinese | WPRIM | ID: wpr-878466

ABSTRACT

To achieve uniform soluble expression of multiple proteins in the same Escherichia coli strain, and simplify the process steps of antigen production in genetic engineering subunit multivalent vaccine, we co-expressed three avian virus proteins including the fowl adenovirus serotype 4 (FAdV-4) Fiber-2 protein, infectious bursal disease virus (IBDV) VP2 protein and egg-drop syndrome virus (EDSV) Fiber protein in E. coli BL21(DE3) cells after optimization of gene codon, promoter, and tandem expression order. The purified proteins were analyzed by Western blotting and agar gel precipitation (AGP). The content of the three proteins were well-proportioned after co-expression and the purity of the purified proteins were more than 80%. Western blotting analysis and AGP experiment results show that all the three co-expression proteins had immunoreactivity and antigenicity. It is the first time to achieve the three different avian virus antigens co-expression and co-purification, which simplified the process of antigen production and laid a foundation for the development of genetic engineering subunit multivalent vaccine.


Subject(s)
Animals , Antigens, Viral/genetics , Biological Assay , Chickens/immunology , Escherichia coli/genetics , Infectious bursal disease virus/immunology , Poultry Diseases , Vaccines, Synthetic/isolation & purification , Viral Structural Proteins/immunology , Viral Vaccines/immunology
7.
Mem. Inst. Oswaldo Cruz ; 115: e200287, 2020. tab, graf
Article in English | LILACS | ID: biblio-1154869

ABSTRACT

BACKGROUND The heat-labile nature of Dengue virus (DENV) in serum samples must be considered when applying routine diagnostic tests to avoid issues that could impact the accuracy of test results with direct implications for case management and disease reporting. OBJECTIVES To check if pre-analytical variables, such as storage time and temperature, have an impact on the accuracy of the main routine diagnostic tests for dengue. METHODS Virus isolation, reverse transcription real-time polymerase chain reaction (RT-PCR) and NS1 enzyme-linked immunosorbent assay (ELISA) were evaluated using 84 samples submitted to different pre-analytical conditions. FINDINGS Sensitivity and negative predictive value were directly affected by sample storage conditions. RT-PCR and virus isolation showed greater dependence on well-conserved samples for an accurate diagnosis. Interestingly, even storage at -30ºC for a relatively short time (15 days) was not adequate for accurate results using virus isolation and RT-PCR tests. On the other hand, NS1 ELISA showed no significant reduction in positivity for aliquots tested under the same conditions as in the previous tests. MAIN CONCLUSIONS Our results support the stability of the NS1 marker in ELISA diagnosis and indicate that the accuracy of routine tests such as virus isolation and RT-PCR is significantly affected by inadequate transport and storage conditions of serum samples.


Subject(s)
Humans , Immunologic Tests/methods , Enzyme-Linked Immunosorbent Assay/methods , Viral Nonstructural Proteins/immunology , Reverse Transcriptase Polymerase Chain Reaction/methods , Dengue/diagnosis , Dengue Virus/isolation & purification , Antigens, Viral/blood , Predictive Value of Tests , Sensitivity and Specificity , Viral Nonstructural Proteins/genetics , Dengue/blood , Dengue/virology , Dengue Virus/genetics , Dengue Virus/immunology , Antibodies, Viral/blood , Antigens, Viral/immunology
8.
Clinics ; 75: e2290, 2020. tab, graf
Article in English | LILACS | ID: biblio-1142772

ABSTRACT

OBJECTIVES: COVID-19 is a public health emergency of international concern whose detection in recovered asymptomatic patients is dependent on accurate diagnosis as it enables the estimation of the susceptibility of the population to the infection. This demand has resulted in the development of several commercial assays employing recombinant proteins, but the results of these assays are not reliable as they do not involve comparison with natural viral antigens. We independently used the SARS-CoV-2 whole viral antigen (WVA) and recombinant nucleocapsid protein (rNP) to develop in-house ELISAs for IgG detection; the results of these ELISAs were then compared to obtain reliable results. METHODS: WVA and rNP ELISAs were performed on COVID-19 negative sera from patients before the pandemic in Brazil, and on RT-qPCR-positive or SARS-CoV-2-IgG against rNP and IgG against WVA-positive samples from recently infected patients in Sao Paulo, Brazil. RESULTS: Both ELISAs detected a large fraction of infected patients but exhibited certain drawbacks. Higher signals and lower numbers of false-negatives were observed in rNP ELISA; however, a higher fraction of false-positives was observed in control groups. A high number of false-negatives was observed with WVA ELISA. Correlating the results of rNP and WVA ELISAs resulted in improved performance for COVID-19 diagnosis. CONCLUSION: The choice of antigen is an important aspect in optimizing the laboratory diagnosis of COVID-19. The use of rNP ELISA for the detection of anti-SARS-CoV-2 IgG antibodies seems promising, but comparison of the results with those of WVA ELISA is crucial for accurate test development prior to commercialization. IgG serology using several assays, and with the spectral patterns of SARS-CoV-2, resulted in confusing information that must be clarified before the establishment of diagnostic serology criteria.


Subject(s)
Humans , SARS-CoV-2 , COVID-19 , Brazil , Sensitivity and Specificity , Clinical Laboratory Techniques , COVID-19 Testing , Antibodies, Viral , Antigens, Viral
9.
Chinese Journal of Biotechnology ; (12): 281-289, 2019.
Article in Chinese | WPRIM | ID: wpr-771378

ABSTRACT

In previous studies, we found that truncated rotavirus VP4* (aa 26-476) could be expressed in soluble form in Escherichia coli and confer high protection against rotavirus in the mouse mode. In this study, we further improved the immunogenicity of VP4* by polymerization. The purified VP4* was polymerized through incubation at 37 ℃ for 24 h, and then the homogeneity of the particles was analyzed by HPLC, TEM and AUC, while the thermal stability and antigenicity was analyzed by DSC and ELISA, respectively. Finally, the immunogenicity and protective efficacy of the polymers analyzed by a mouse maternal antibody model. The results showed that VP4* aggregated into homogeneous polymers, with high thermostability and neutralizing antibody binding activity. In addition, VP4* polymers (endotoxin <20 EU/dose) stimulated higher neutralizing antibodies and confer higher protection against rotavirus-induced diarrhoea compared with the VP4* trimers when immunized with aluminium adjuvant. In summary, the study in VP4* polymers provides a new strategy for the development of recombinant rotavirus vaccines.


Subject(s)
Animals , Antibodies, Viral , Antigens, Viral , Capsid , Capsid Proteins , Mice , Polymerization , Rotavirus , Rotavirus Infections
10.
Rev. Soc. Bras. Med. Trop ; 52: e20180353, 2019. tab, graf
Article in English | LILACS | ID: biblio-1057248

ABSTRACT

Abstract INTRODUCTION: Dengue is an important mosquito-borne disease in tropical and subtropical regions. Adhesion molecules have not been systematically characterized in the renal tissue of patients with severe dengue (SD). The objective of this study was to detect viral antigens in samples from patients that evolved with SD, correlating with the expression of ICAM-1, VCAM-1, VE-cadherin, and E-selectin to contribute to a better understanding of the pathophysiology of SD. METHODS: Kidney specimens from patients with SD were selected according to clinical and laboratorial data and submitted to histological and immunohistochemistry analysis. A semiquantitative evaluation was performed considering positive immunostaining in 20 glomeruli. RESULTS: Viral antigens were mainly detected in distal tubules. The intense immunostaining of VCAM-1 and ICAM-1 was observed. The expression of E-selectin was discrete, and VE-cadherin expression varied from mild to moderate. VCAM-1 was slightly intense in the glomerular capsule; the expression of ICAM-1 was diffuse. E-selectin was diffuse, and VE-cadherin varied from mild to moderate. The most frequent histological findings were glomerular congestion, mild glomerulitis, acute renal injury, and glomerular atrophy. CONCLUSIONS: The results appear to demonstrate an imbalance between vascular endothelial permeability regulating events in renal lesions in SD. The increase in the expression of ICAM-1 and VCAM-1 is an in-situ indicator of higher permeability with a consequent influx of cells favoring the inflammation of the endothelium. These molecules are important in the pathophysiology of the disease and provide the possibility of developing new markers for the evaluation, clinical follow-up, and therapeutic response of patients with SD.


Subject(s)
Humans , Male , Female , Child, Preschool , Child , Adolescent , Adult , Young Adult , Intercellular Adhesion Molecule-1/physiology , Vascular Cell Adhesion Molecule-1/physiology , E-Selectin/physiology , Severe Dengue/physiopathology , Severe Dengue/blood , Endothelium/physiopathology , Immunohistochemistry , Biomarkers/blood , Antigens, CD/physiology , Antigens, CD/blood , Cadherins/physiology , Cadherins/blood , Up-Regulation , Intercellular Adhesion Molecule-1/blood , Disease Progression , Vascular Cell Adhesion Molecule-1/blood , E-Selectin/blood , Middle Aged , Antigens, Viral/blood
11.
Rev. Soc. Bras. Med. Trop ; 52: e20180457, 2019. tab, graf
Article in English | LILACS | ID: biblio-1041557

ABSTRACT

Abstract INTRODUCTION: We defined the cut-off values of the antigenemia and cytomegalovirus (CMV) DNA tests in HIV/AIDS patients to identify CMV disease. METHODS: A total of 97 samples from 68 patients with and without CMV disease were analyzed by viral DNA detection and antigenemia assay. RESULTS: Qualitative and quantitative results significantly differed between assays. The cut-off values for the antigenemia and qPCR assays were 1.5 positive cells/200,000 leukocytes and 3.715 log/mL, respectively. CONCLUSIONS: Antigenemia and qPCR are suitable for monitoring CMV disease in HIV patients, however, the threshold values should be determined within the centers where the patients are monitored.


Subject(s)
Humans , DNA, Viral/analysis , AIDS-Related Opportunistic Infections/diagnosis , Cytomegalovirus Infections/diagnosis , Cytomegalovirus/isolation & purification , Brazil/epidemiology , DNA, Viral/blood , Predictive Value of Tests , Prospective Studies , ROC Curve , Sensitivity and Specificity , AIDS-Related Opportunistic Infections/blood , Cytomegalovirus Infections/blood , Viral Load , Cytomegalovirus/genetics , Real-Time Polymerase Chain Reaction , Antigens, Viral/blood
12.
Article in Korean | WPRIM | ID: wpr-714781

ABSTRACT

As part of the immunoserology program of the Korean Association of External Quality Assessment Service, we organized two trials on the external quality assessment of hepatitis viral markers in 2016 and 2017. The hepatitis viral antigens and antibodies program consisted of 10 test items. We delivered two and three types of pooled sera specimens to 965 and 965 institutions for the first and second trials of external proficiency testing in 2016, respectively. The number of participating laboratories was 915 (94.8%) and 913 (95.0%) in the first and second trials in 2016, respectively. We also delivered three kinds of pooled sera specimens to 936 and 1,015 institutions for the first and second trials of external proficiency testing in 2017, respectively. The number of participating laboratories was 920 (98.3%) and 996 (98.1%) in the first and second trials in 2017, respectively. The most commonly tested items were hepatitis B surface antigen, followed by the antibodies to hepatitis B surface antigen, anti-hepatitis C virus, hepatitis B envelope antigen, antibodies to hepatitis B envelope antigen, anti-hepatitis A virus and antibodies to hepatitis B core antigen. The most frequently used methods for detecting viral markers were the chemiluminescence immunoassay and the electrochemiluminescence immunoassay, but they yielded a few-false positive results due to the matrix effect. The immunochromatographic assay yielded false-negative results for anti-hepatitis A virus due to low sensitivity. Continuous improvement in the quality of viral hepatitis testing through participation in the survey seems necessary.


Subject(s)
Antibodies , Antigens, Viral , Biomarkers , Hepatitis A , Hepatitis B , Hepatitis B Core Antigens , Hepatitis B Surface Antigens , Hepatitis B virus , Hepatitis C , Hepatitis , Immunoassay , Chromatography, Affinity , Korea , Laboratory Proficiency Testing , Luminescence
13.
Arq. neuropsiquiatr ; 75(8): 580-588, Aug. 2017. tab, graf
Article in English | LILACS | ID: biblio-888309

ABSTRACT

ABSTRACT The polyspecific antibody synthesis in multiple sclerosis (MS) gained diagnostic relevance with the frequent combination of measles-, rubella- and varicella zoster antibodies (MRZ-antibody reaction) but their pathophysiological role remains unknown. This review connects the data for intrathecal polyspecific antibody synthesis in MS and neurolupus with observations in the blood of patients with Guillain-Barré syndrome (GBS). Simultaneously increased antibody and autoantibody titers in GBS blood samples indicate that the polyspecific antibodies are based on a general property of an immune network, supported by the deterministic day-to-day concentration variation of antibodies in normal blood. Strongly correlated measles- and rubella- antibody variations point to a particular connectivity between the MRZ antibodies. The immune network, which provides serological memory in the absence of an antigen, implements the continuous change of the MRZ pattern in blood, not followed by the earlier immigrated B cells without corresponding connectivity in the brain. This may explain the different antibody patterns in cerebrospinal fluid, aqueous humor and blood of the individual MS patient. A complexity approach must implement a different view on causation in chronic diseases and causal therapies.


RESUMO A síntese de anticorpos poliespecíficos em esclerose múltipla (EM) ganhou relevância diagnóstica com a combinação frequente de anticorpos contra sarampo, rubéola e varicela-zoster (reação de anticorpos MRZ), mas seu papel fisiopatológico permanece desconhecido. Esta revisão relaciona os dados da síntese intratecal de anticorpos poliespecíficos em EM e Neurolupus com observações no sangue de pacientes com síndrome de Guillain Barré (SGB). Simultaneamente, os títulos aumentados de anticorpos e autoanticorpos em amostras de sangue de SGB indicam que os anticorpos poliespecíficos se baseiam numa propriedade geral de uma rede imunitária, suportada pela variação determinística da concentração diária de anticorpos no sangue normal. As variações fortemente correlacionadas de anticorpos contra sarampo e rubéola apontam para uma conectividade particular entre os anticorpos MRZ. A rede imunitária, que fornece memória sorológica na ausência de um antígeno, implementa a mudança contínua do padrão MRZ no sangue, não seguida pelas células B que imigraram anteriormente sem conectividade no cérebro. Isto pode explicar os diferentes padrões de anticorpos no LCR, humor aquoso e sangue do paciente individual de EM. Uma abordagem complexa deve implementar uma visão diferente sobre a causalidade em doenças crônicas e terapias causais.


Subject(s)
Humans , Guillain-Barre Syndrome/immunology , Antibodies, Viral/blood , Multiple Sclerosis/immunology , Antibody Specificity/immunology , Rubella/immunology , Immunoglobulin G/blood , Cerebrospinal Fluid/chemistry , Herpes Zoster/immunology , Measles/immunology , Antibodies, Bacterial , Multiple Sclerosis/cerebrospinal fluid , Mumps/immunology , Antigens, Viral/immunology
14.
J. pediatr. (Rio J.) ; 93(3): 246-252, May.-June 2017. tab, graf
Article in English | LILACS | ID: biblio-841353

ABSTRACT

Abstract Objective: The aim of this study was to evaluate the QuickVue® RSV Test Kit (QUIDEL Corp, CA, USA) as a screening tool for respiratory syncytial virus in children with acute respiratory disease in comparison with the indirect immunofluorescence assay as gold standard. In Brazil, rapid antigen detection tests for respiratory syncytial virus are not routinely utilized as a diagnostic tool, except for the diagnosis of dengue and influenza. Methods: The authors retrospectively analyzed 486 nasopharyngeal aspirate samples from children under age 5 with acute respiratory infection, between December 2013 and August 2014, the samples were analyzed by indirect immunofluorescence assay and QuickVue® RSV Test kit. Samples with discordant results were analyzed by real time PCR and nucleotide sequencing. Results: From 313 positive samples by immunofluorescence assays, 282 (90%) were also positive by the rapid antigen detection test, two were positive only by rapid antigen detection test, 33 were positive only by immunofluorescence assays, and 171 were positive by both methods. The 35 samples with discordant results were analyzed by real time PCR; the two samples positive only by rapid antigen detection test and the five positive only by immunofluorescence assays were also positive by real time PCR. There was no relation between the negativity by QuickVue® RSV Test and viral load or specific strain. The QuickVue® RSV Test showed sensitivity of 90%, specificity of 98.8%, predictive positive value of 99.3%, and negative predictive value of 94.6%, with accuracy of 93.2% and agreement κ index of 0.85 in comparison to immunofluorescence assay. Conclusions: This study demonstrated that the QuickVue® RSV Test Kit can be effective in early detection of Respiratory syncytial virus in nasopharyngeal aspirate and is reliable for use as a diagnostic tool in pediatrics.


Resumo Objetivo: Avaliar o teste QuickVue® RSV Test Kit (QUIDEL Corp, CA, EUA) para o diagnóstico rápido do vírus sincicial respiratório em crianças com doença respiratória aguda, comparandoo com a imunofluorescência indireta como padrão ouro. Visto que, no Brasil, testes rápidos para detecção de antígenos para vírus sincicial respiratório não são rotineiramente utilizados como ferramenta de diagnóstico, exceto para Dengue e Influenza. Métodos: Um total de 486 amostras de aspirado de nasofaringe de crianças menores de 5 anos com doença respiratória aguda, coletadas entre dezembro de 2013 e agosto de 2014, foram analisadas por imunofluorescência e pelo teste QuickVue®. Amostras com resultados discordantes entre os métodos foram submetidas a PCR em tempo real e sequenciamento. Resultados: Das 313 amostras positivas por IFI, 282 foram positivas no teste rápido (90%), 2 amostras foram positivas apenas no teste rápido (0.6%), 33 apenas na imunofluorescência (10.5%) e 171 foram negativas em ambos os métodos. As 35 amostras com resultados discordantes foram testadas por PCR em tempo real, sendo que duas que foram positivas apenas no teste rápido e 5 apenas na imunofluorescência confirmaram-se positivas. Não houve relação entre a ausência de positividade no teste QuickVue® com a carga ou com a cepa viral. O teste QuickVue® mostrou sensibilidade de 90.1%, especificidade 98.9%, valor preditivo positivo 99.3%, valor preditivo negativo de 94.6%, acurácia de 93.2% e índice de concordância de 0.85 em comparação à imunofluorescência. Conclusões: Nosso estudo demonstrou que o teste QuickVue® RSV pode ser efetivo na detecção precoce do vírus sincicial respiratório em amostras de aspirado de nasofaringe e é confiável como uma ferramenta de diagnósticos em pediatria.


Subject(s)
Humans , Male , Female , Child, Preschool , Respiratory Syncytial Viruses/immunology , Respiratory Syncytial Virus Infections/diagnosis , Reverse Transcriptase Polymerase Chain Reaction , Antigens, Viral/analysis , Reagent Kits, Diagnostic , Respiratory Syncytial Viruses/isolation & purification , Respiratory Tract Infections/virology , Brazil , Retrospective Studies , Sensitivity and Specificity , Respiratory Syncytial Virus Infections/virology , Fluorescent Antibody Technique, Indirect
15.
Mem. Inst. Oswaldo Cruz ; 112(6): 458-468, June 2017. tab, graf
Article in English | LILACS | ID: biblio-841802

ABSTRACT

ABSTRACT BACKGROUND Dengue fever may present hemorrhages and cavitary effusions as result of exacerbated immune responses. We investigated hydro-alcoholic extracts from leaves (UGL) and bark (UGB) of the medicinal species Uncaria guinanensis with respect to antiviral effects in Dengue virus (DENV) infection and in immunological parameters associated with in vivo physiopathological features. METHODS Chemical profiles from UGB or UGL were compared in thin layer chromatography and 1H nuclear magnetic resonance using flavonoid compounds and a pentacyclic oxindole alkaloid-enriched fraction as references. DENV-2-infected hepatocytes (Huh-7) were treated with extracts. Cell viability, DENV antigens and immunological factors were detected by enzyme-linked immunosorbent assay (ELISA) or flow cytometry. FINDINGS The UGL mainly differed from UGB by selectively containing the flavonoid kaempferitrin. UGB and UGL improved hepatocyte viability. Both extracts reduced intracellular viral antigen and inhibited the secretion of viral non-structural protein (NS1), which is indicative of viral replication. Reduction in secretion of macrophage migration inhibitory factor was achieved by UGB, of interleukin-6 by UGL, and of interleukin-8 by both UGB and UGL. MAIN CONCLUSIONS The U. guianensis extracts presented, antiviral and immunomodulatory effects for DENV and possibly a hepatocyte-protective activity. Further studies may be performed to consider these products as potential candidates for the development of an herbal product for the future treatment of dengue.


Subject(s)
Humans , Antiviral Agents/pharmacology , Plant Extracts/pharmacology , Cell Survival/drug effects , Cytokines/drug effects , Cytokines/immunology , Chemokines/drug effects , Chemokines/immunology , Uncaria/chemistry , Dengue/physiopathology , Dengue/immunology , Dengue/virology , Dengue Virus/drug effects , Dengue Virus/immunology , Antigens, Viral/drug effects , Antigens, Viral/immunology , Enzyme-Linked Immunosorbent Assay , Flow Cytometry
16.
Mem. Inst. Invest. Cienc. Salud (Impr.) ; 15(1): 7-15, abr. 2017. ilus
Article in Spanish | LILACS, BDNPAR | ID: biblio-1008720

ABSTRACT

Los flavivirus son responsables de una considerable morbi-mortalidad a nivel mundial. Entre ellos, el virus del dengue (DENV) es causante de graves problemas de salud pública en Paraguay. El objetivo del estudio fue detectar infecciones por flavivirus a través de una reacción de RT-nested PCR genérica para flavivirus en 195 muestras de individuos con sospecha de dengue, negativos por el test inmunocromatográfico (antígeno NS1 ­ DENV), provenientes del área metropolitana de Asunción entre 2011 y 2013. Las muestras positivas para flavivirus fueron sometidas a dos reacciones de RT-nested PCRs específicas para DENV. El límite de detección (LD) para flavivirus fue de 0,2 UFP/reacción. En total 43/195 muestras fueron positivas para flavivirus. De estas, 38/43 (88,4%) correspondieron a DENV (6 DENV-1, 30 DENV-2 y 2 DENV-3). Además, 5/43 casos (11,6%) positivos para flavivirus fueron negativos para DENV por ambas reacciones específicas, pudiendo deberse a infecciones por otros flavivirus. Los resultados sugieren que la utilización de una reacción genérica seguida de otras reacciones específicas para DENV en casos febriles negativos para NS1 por el método inmunocromatográfico permitiría detectar más casos de infecciones por DENV y además, podría contribuir a la identificación de casos debido a infecciones por otros flavivirus.


Flaviviruses are responsible for considerable worldwide morbidity and mortality. Among them, the dengue virus (DENV) causes serious public health problems in Paraguay. The objective of the study was to detect flavivirus infections using a generic RT-nested -PCR in 195 samples of individuals with suspected dengue and negative for the inmunochromatographic test (NS1 antigen ­ DENV), from the metropolitan area of Asuncion between 2011 and 2013. The flavivirus-positive samples were subjected to two reactions of DENV-specific RT-nested PCRs. The detection limit (DL) for flavivirus was 0.2 PFU / reaction. In total, 43/195 samples were positive for flavivirus. Of them, 38/43 (88,4%) corresponded to DENV (6 DENV-1, 30 DENV-2 and 2 DENV-3). In addition, 5/43 cases (11.6%) positive for flavivirus were negative for DENV by both specific reactions, and may be infections caused by other flaviviruses. The results suggest that the use of a generic reaction followed by other DENV specific reactions in febrile negative cases for NS1 by the immunochromatographic method would allow the detection of more cases of DENV infections and could contribute to the identification of cases due to infections by others flaviviruses.


Subject(s)
Humans , Male , Female , Infant , Child, Preschool , Child , Adolescent , Adult , Middle Aged , Aged , Aged, 80 and over , Flavivirus Infections/diagnosis , Dengue Virus/isolation & purification , Flavivirus/isolation & purification , Paraguay , Cross-Sectional Studies , Genome, Viral , Reverse Transcriptase Polymerase Chain Reaction , Dengue Virus/genetics , Dengue Virus/immunology , Fever , Flavivirus/genetics , Antigens, Viral/isolation & purification
17.
Article in Chinese | WPRIM | ID: wpr-297247

ABSTRACT

<p><b>OBJECTIVE</b>To investigate the antigen clearance time, time to symptom disappearance, and the association between them using immunofluorescence assay for dynamic monitoring of influenza virus antigen in children with influenza.</p><p><b>METHODS</b>A total of 1 063 children suspected of influenza who visited the Hunan People's Hospital from March to April, 2016 were enrolled. The influenza A/B virus antigen detection kit (immunofluorescence assay) was used for influenza virus antigen detection. The children with positive results were given oseltamivir as the antiviral therapy and were asked to re-examine influenza virus antigen at 5, 5-7, and 7 days after onset.</p><p><b>RESULTS</b>Of all children suspected of influenza, 560 (52.68%) had an influenza virus infection. A total of 215 children with influenza virus infection were followed up. The clearance rate of influenza virus antigen was 9.8% (21 cases) within 5 days after onset. The cumulative clearance rate of influenza virus antigen was 32.1% (69 cases) within 5-7 days, and 98.1% (211 cases) within 7-10 days after onset. Among these children, 6 children (2.8%) achieved the improvement in clinical symptoms within 3 days after onset. The cumulative rate of symptom improvement was 84.7% (182 cases) within 3-5 days after onset, and 100% achieved the improvement after 5 days of onset.</p><p><b>CONCLUSIONS</b>The time to improvement in symptoms after treatment is earlier than antigen clearance time. Almost all of the children achieve influenza virus antigen clearance 7-10 days after onset. Therefore, it is relatively safe for children to go back to school within 7-10 days after onset when symptoms disappear.</p>


Subject(s)
Antigens, Viral , Blood , Child , Child, Preschool , Female , Fluorescent Antibody Technique , Humans , Infant , Influenza A virus , Allergy and Immunology , Influenza B virus , Allergy and Immunology , Male , Time Factors
18.
Article in Chinese | WPRIM | ID: wpr-297227

ABSTRACT

<p><b>OBJECTIVE</b>To investigate the expression of IFN-λ1 in respiratory epithelial cells of children with respiratory syncytial virus (RSV) infection and its relationship with RSV load.</p><p><b>METHODS</b>The nasopharyngeal swabs were collected from the children who were hospitalized with respiratory tract infection from June 2015 to June 2016. A direct immunofluorescence assay was used to detect the antigens of seven common respiratory viruses (including RSV) in the nasopharyngeal swabs. A total of 120 children who were only RSV positive were selected as the RSV infection group. A total of 50 children who had negative results in the detection of all viral antigens were selected as the healthy control group. Fluorescence quantitative real-time PCR was used to determine the RSV load and the expression of IFN-λ1 mRNA in the nasopharyngeal swabs of children in the two groups.</p><p><b>RESULTS</b>The expression of IFN-λ1 in the RSV infection group was significantly higher than that in the healthy control group (P<0.05). The expression of IFN-λ1 was positively correlated with RSV load (r=0.56, P<0.05).</p><p><b>CONCLUSIONS</b>RSV can induce the expression of IFN-λ1 in respiratory epithelial cells, suggesting that IFN-λ1 may play an important role in anti-RSV infection.</p>


Subject(s)
Antigens, Viral , Child, Preschool , Epithelial Cells , Allergy and Immunology , Female , Humans , Infant , Infant, Newborn , Interleukins , Physiology , Male , Nasopharynx , Microbiology , Real-Time Polymerase Chain Reaction , Respiratory Syncytial Virus Infections , Allergy and Immunology , Virology , Viral Load
19.
VozAndes ; 28(1): 47-48, 2017.
Article in Spanish | LILACS | ID: biblio-986907

ABSTRACT

Como primer paso para abordar el manejo de rehidratación oral se debe realizar una estratifcación de la severidad de la deshidratación utilizando preferentemente el "método Dhaka" [5]; tabla 1. El défcit de líquidos puede estimarse mediante la diferencia del peso corporal entre los momentos previos y al momento de la atención (relación 1 Kg equivale a 1 litro). Se recomienda iniciar la hidratación preferentemente por vía oral; reponiendo primero las pérdidas producidas por las deposiciones y luego dar un mantenimiento de los líquidos. Para adultos, las sales de rehidratación oral (SRO) se administrarían a dosis de 50-100 mL/Kg cada 4 a 6 horas, ingeridas en sorbos pequeños y entre 200 y 400 mL luego de cada nueva deposición. Debe considerarse también la posibilidad de administrar SRO por sonda nasogástrica para casos de deshidratación severa cuando la rehidratación intravenosa no sea posible hasta llegar a un centro con las facilidades necesarias.


Subject(s)
Humans , Rehydration Solutions , Dysentery , Antigens, Viral , Adult
20.
Chinese Journal of Virology ; (6): 195-202, 2016.
Article in Chinese | WPRIM | ID: wpr-296197

ABSTRACT

In order to develop a combined live vaccine that will be used to prevent against porcine parvovirus (PPV) and Pseudorabies virus (PRV) infection, the VP2 gene of PPV was inserted into the transfer vector plasmid pG to produce the recombinant plasmid pGVP2. The plasmid pGVP2 and the genome of PRV HB98 attenuated vaccine were transfected by using lipofectamine into swine testis cells for the homologous recombination. The recombinant virus rPRV-VP2 was purified by selection of green fluorescence plaques for five cycles. 6-week-old female Kunming mice were immunized intramuscularly with attenuated PRV parent HB98 strain, commercial inactivated vaccine against PPV, recombinant virus, DMEM culture solution. The injections were repeated with an equivalent dose after 2 weeks in all of the groups, and then challenged with the virulent PRV NY strain at 7 weeks after the first immunization. The recombinant virus rPRV-VP2 was successfully generated, and the recombinant virus could effectively elicite anti-PPV and PRV antibody and significant cellular immune response as indicated by anti-PPV ELISA and HI, PRV-neutralizing assay and flow cytometry. The challenge assay indicated that recombinant virus could protect the mice against the virulent PRV challenge. These results demonstrated that the recombinant virus can be a candidate recombinant vaccine strain for the prevention of PRV and PPV.


Subject(s)
Animals , Antibodies, Viral , Allergy and Immunology , Antigens, Viral , Genetics , Allergy and Immunology , Capsid Proteins , Genetics , Allergy and Immunology , Female , Gene Expression , Genetic Vectors , Genetics , Metabolism , Herpesvirus 1, Suid , Genetics , Metabolism , Mice , Parvovirus, Porcine , Genetics , Allergy and Immunology , Swine , Swine Diseases , Allergy and Immunology , Virology , Viral Vaccines , Genetics , Allergy and Immunology
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