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1.
Biomed Microdevices ; 20(3): 78, 2018 09 04.
Artigo em Inglês | MEDLINE | ID: mdl-30182296

RESUMO

Dengue is an infectious viral disease transmitted by the Aedes aegypti mosquito, the control of which is complex. In addition, the clinical diagnosis is difficult to perform since it resembles other febrile infections; thus, the development of more effective methods to detect dengue virus (DV) has drawn increasing attention. The present study aimed to develop an impedimetric immunosensor for dengue diagnosis using a screen-printed electrode (SPE) functionalized with polymer films derived from 4-aminophenylacetic acid (4-APA). Data obtained from scanning electron microscopy (SEM) showed the deposition of a uniformly distributed material over the electrode surface. The immunosensor was based on the specific interaction between dengue antigen, NS1 protein, and anti-NS1 antibodies, IgG and IgM. In a characterization study using cyclic voltammetry (CV), the polymer film showed two oxidation peaks at +0.17 and + 0.35 V in 0.50 M sulfuric acid solution, indicating its adsorption and electroactivity at the SPE surface. Electrochemical impedance spectroscopy (EIS) measurements showed a higher charge transfer resistance (Rct) to the polymer film-modified SPE as compared with the bare SPE, corroborating a previous study. The best rNS1 concentration for immobilization was 1.00 ng/mL, and the immunoreaction time between the antigen (Ag) and the antibody (Ab) was 20 min. Dilutions of positive and negative clinical serum samples were evaluated by EIE, from which it was possible to elucidate, for the positive serum, that the more diluted the serum the greater the Rct. Negative serum also showed an analytical signal, probably due to the presence of non-specific antibodies; however, the generated signal presented values closer to the rNS1 signal, indicating good selectivity of the proposed platform. The experiments were repeated using bare SPE to verify the importance of the polymer film in biosensor construction. No significant difference was observed between these results. Graphical abstract Proposed schematic for the genosensor development.


Assuntos
Compostos de Anilina/química , Técnicas Biossensoriais , Materiais Revestidos Biocompatíveis/química , Vírus da Dengue/isolamento & purificação , Dengue/diagnóstico , Grafite/química , Fenilacetatos/química , Anticorpos Antivirais/sangue , Antígenos Virais/sangue , Dengue/sangue , Dengue/imunologia , Vírus da Dengue/imunologia , Espectroscopia Dielétrica , Eletrodos , Humanos , Limite de Detecção , Polímeros/química , Propriedades de Superfície , Proteínas não Estruturais Virais/metabolismo
2.
J Clin Lab Anal ; 32(6): e22410, 2018 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-29453831

RESUMO

BACKGROUND: The most of the hepatitis C-infected patients remain undiagnosed until they develop severe liver damage or submitted for serological screening. OBJECTIVE: To evaluate a recombinant multiepitope protein for detection of IgG anti-hepatitis C virus. METHOD: A synthetic gene was cloned, expressed in Escherichia coli, and the recombinant protein was purified. Human serum panel consisted of 88 positives (20 HCV genotyped) and 376 negatives for hepatitis C, 6 positives for human acquired immunodeficiency virus, 6 syphilis positives, 6 hepatitis B positives were tested by IgG antihepatitis C virus using the protein by enzyme-linked immunosorbent assay. In addition, 20 positive (all genotyped samples) and 20 negative samples were also tested by immunoblot and dot blot assays. RESULTS: Positive hepatitis C sera were strongly reactive against the protein by immunoblot assay. In the dot blot assay, positive sera were reactive until 1:1000 dilution and there were no false positive results in the hepatitis C negative sera. In the enzyme-linked immunosorbent assay, positive and negative sera had significant discrimination. No cross-reaction was observed in samples positive for syphilis; human acquired immunodeficiency virus and hepatitis B. All 20 genotyped samples were positive by the three methods. CONCLUSION: The multiepitope protein used here has a lower cost compared to production of each antigen separately and could be an alternative for the serological diagnosis of hepatitis C.

3.
Rejuvenation Res ; 27(2): 44-50, 2024 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-38279807

RESUMO

Despite current literature pointing to a link between shortened telomeres and aging, chronic diseases, and geriatric syndromes, the precise implications of this connection remain unclear. The aim of this exploratory, cross-sectional, observational study was to investigate the association between the relative telomere length (RTL) of peripheral blood leukocyte subtypes (mononuclear cells and granulocytes) and physical performance using the Short Physical Performance Battery (SPPB) in older adults. A cohort of 95 participants was recruited, which included men and women aged over 60 years (70.48 ± 5.5 years). It was found that mononuclear cell RTL was significantly lower than that of granulocytes (p < 0.0001). Moreover, individuals with good SPPB performance exhibited lower mononuclear cell RTL compared with those with moderate or poor performance. However, no significant differences were observed in granulocyte RTL between different SPPB performance groups. The global SPPB score showed an inverse correlation with mononuclear cell RTL, but this correlation was not present with granulocyte RTL. Similarly, the SPPB sit-to-stand domain correlated with mononuclear cell RTL, but no such correlation was found with granulocyte RTL. Our findings challenge conventional expectations, suggesting that shorter mononuclear cell RTL may be associated with favorable functional capacity. The variations in RTL between mononuclear cells and granulocytes highlight their distinct biological roles and turnover rates. A history of immune responses may influence mononuclear cell RTL dynamics, while telomerase activity may protect granulocyte RTL from significant shortening. The unexpected associations observed in mononuclear cell RTL emphasize the complex interplay between immune responses, cellular aging, and functional capacity in older adults.


Assuntos
Envelhecimento , Leucócitos , Masculino , Humanos , Feminino , Idoso , Pessoa de Meia-Idade , Estudos Transversais , Encurtamento do Telômero , Telômero , Desempenho Físico Funcional
4.
Sci Rep ; 13(1): 1553, 2023 01 27.
Artigo em Inglês | MEDLINE | ID: mdl-36707661

RESUMO

Certain cut-off points for sarcopenia screening and diagnosis are arbitrary and based on European populations, with normative references often obtained from healthy young adults. Although respiratory skeletal muscle strength tests represent low-cost clinical measures commonly performed in clinical practice by health professionals, a gap remains regarding whether respiratory skeletal muscle strength tests are adequate and sensitive measures for sarcopenia screening. This study aimed to verify the value of handgrip and respiratory muscle strength as possible discriminators to identify sarcopenia and to establish cut-off points for sarcopenia screening in community-dwelling, Brazilian women. In a cross-sectional study, 154 community-dwelling, Brazilian women (65-96 years) were assessed for appendicular skeletal muscle mass, handgrip (HGS), and respiratory muscular strength, including maximal inspiratory pressure (MIP) and maximal expiratory pressure (MEP). The data were analyzed using the ROC curve and the Youden Index determined cut-off points. Statistical significance was set at 5%. 88 participants (57%) were sarcopenic. MEP (OR 0.98 [95%CI 0.97, 1.00], p = 0.023) and HGS (OR 0.82 [95% CI 0.75, 0.90], p < 0.001) were independent factors for sarcopenia in older. The optimal cut-off points for identifying sarcopenia were ≤ 77 cmH2O for MEP (AUC = 0.72), and ≤ 20 kg for HGS (AUC = 0.80). Simple muscular strength tests, including HGS and MEP, may be considered in the identification of sarcopenia in older, community-dwelling, Brazilian women. Future work is still needed to assess external validation of the proposed cut-offs before the clinical application.


Assuntos
Sarcopenia , Adulto Jovem , Humanos , Feminino , Idoso , Sarcopenia/diagnóstico , Força da Mão/fisiologia , Vida Independente , Brasil , Estudos Transversais , Força Muscular/fisiologia , Músculo Esquelético , Músculos Respiratórios
5.
J Vasc Access ; 22(2): 209-217, 2021 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-32588732

RESUMO

BACKGROUND: Central venous catheter-related bloodstream infection is an important adverse event in health care. Molecular methods are not yet substitutive of microbiological in the detection of the pathogens responsible for the infection, but they can help in the epidemiological characterization. AIM: To detect bacteria by polymerase chain reaction, from material extracted from the tip of central catheters of patients suspected of infection at the intensive care unit. METHODS: Catheters (n = 34) of patients suspected of central venous catheter-related infection were analyzed by polymerase chain reaction. The findings were compared with culture of catheter tip and blood cultures performed by the hospital. FINDINGS: The prevalence of bacteria was Staphylococcus aureus (50%), Enterococcus faecalis (41.2%), Klebsiella pneumoniae (32.4), Pseudomonas aeruginosa (20.6%), Acinetobacter baumannii (38.2%), Escherichia coli (2.9%), and Enterobacter cloacae (0%). No blood culture showed bacterial growth, the culture of catheter tip revealed bacteria in 21 (61.8%) and the polymerase chain reaction had positivity in 31 (91.2%) of the catheters. The mean central venous catheter time was 11 days, and the jugular vein was the site of insertion. CONCLUSION: The molecular method identified more bacteria than microbiological methods and revealed colonization of the catheters. The most commonly found bacteria are in the environment and in the microbiota of the skin, which suggests contamination by the hands of health professionals and points out the need for more efforts in preventive strategies.


Assuntos
Bacteriemia/microbiologia , Bactérias/genética , Técnicas Bacteriológicas , Infecções Relacionadas a Cateter/microbiologia , Cateterismo Venoso Central , Cateteres de Demora/microbiologia , Cateteres Venosos Centrais/microbiologia , Reação em Cadeia da Polimerase , Adolescente , Adulto , Bacteriemia/diagnóstico , Bactérias/isolamento & purificação , Infecções Relacionadas a Cateter/diagnóstico , Cateterismo Venoso Central/efeitos adversos , Cateterismo Venoso Central/instrumentação , Estudos Transversais , Feminino , Hospitalização , Humanos , Pacientes Internados , Unidades de Terapia Intensiva , Masculino , Pessoa de Meia-Idade , Valor Preditivo dos Testes , Ribotipagem , Adulto Jovem
6.
Rev Soc Bras Med Trop ; 53: e20190181, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32187333

RESUMO

INTRODUCTION: Human herpesvirus (HHV)-7 establishes a latent infection during the lifetime of the host and can reactivate after the primary infection, leading to lytic replication in immunosuppressed patients. METHODS: This study aimed to develop an enzyme-linked immunosorbent assay (ELISA) to identify HHV-7 serum antibodies and compare its performance with that of an indirect immunofluorescence assay (IFA). RESULTS: Serum samples (n=102) were tested by IgG-IFA and by ELISA. IFA and ELISA showed IgG-positive results in 77 and 73 samples, respectively. Qualitative concordance of 96% was demonstrated between the two techniques. CONCLUSIONS: ELISA may be useful to diagnose HHV-7 infection.


Assuntos
Anticorpos Antivirais/sangue , Herpesvirus Humano 7/imunologia , Imunoglobulina G/sangue , Infecções por Roseolovirus/diagnóstico , Ensaio de Imunoadsorção Enzimática , Técnica Indireta de Fluorescência para Anticorpo , Humanos , Curva ROC , Sensibilidade e Especificidade
7.
Rejuvenation Res ; 23(5): 387-393, 2020 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-31992162

RESUMO

Leukocyte telomere length in the elderly has been positively associated with healthy living and physical activity. Factors that interfere with telomere shortening are similar to those that may be associated with decreasing functional capacity. To investigate the relationship between mean leukocyte telomere length and functional capacity in community-dwelling elderly individuals, this is an observational, cross sectional, multicentric study conducted with elderly Brazilian patients. Sample characterization was performed using a sociodemographic clinical questionnaire. Telomere length was evaluated by quantitative polymerase chain reaction, and functional capacity was evaluated by the Short Physical Performance Battery (SPPB). A total of 113 elderly individuals (age 70 ± 5.4 years; 75 women and 38 men) were enrolled in this study. Unexpectedly, it was found that lower relative telomere length was associated with better physical capacity in the global SPPB score. Although telomere shortening is observed with increasing age, it is not associated with decreased functional capacity. Functionality is broad and multidimensional, involving the connection of biopsychosocial and cultural factors. While functionality may not be considered a marker of functional aging in an elderly cohort, it can still play an important role in longitudinal studies, which attempt to elucidate process theories. Future studies should use different techniques to measure telomere lengths in subpopulations of cells.


Assuntos
Avaliação Geriátrica , Encurtamento do Telômero , Telômero , Idoso , Envelhecimento , Brasil , Estudos Transversais , Feminino , Humanos , Leucócitos , Masculino
8.
Rejuvenation Res ; 21(4): 294-303, 2018 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-28482745

RESUMO

We examined associations between adverse childhood experiences (ACEs) and shorter telomere length (TL) in 83 older women, including 42 women with less than secondary education and 41 with secondary or more education in a city of Northeast Brazil, a region with substantial socioeconomic inequalities. The low education sample was selected from a representative survey at local neighborhood health centers, while the high education group consisted of a convenience sample recruited by advertising in community centers and centers affiliated with the local university. Relative leukocyte TL was measured by quantitative polymerase chain reaction from blood samples. ACEs were self-reported. Spline linear regression was fitted to assess the strength of the associations between ACEs and TL. Among women with low education, median TL was 1.02 compared with 0.64 in the high education group (p = 0.0001). Natural log-transformed T/S ratio as the dependent variable was used in analysis. Women with low education had been exposed to more ACEs, and among them those experiencing two or more ACEs had longer TL than women exposed to ≤1 ACEs (p = 0.03); among women with high education, this difference was not significant (p = 0.49). In analyses adjusted by age, education, and parental abuse of alcohol, the linear trend of higher TL with increasing ACEs was confirmed (p = 0.02), and the mean difference in TL between groups remained significant (p = 0.002). The unexpected positive relationship between low education and ACEs with TL suggests that older adults who have survived harsh conditions prevailing in Northeast Brazil have the longest TL of their birth cohort.


Assuntos
Acontecimentos que Mudam a Vida , Encurtamento do Telômero/genética , Idoso , Alcoolismo/patologia , Brasil , Escolaridade , Feminino , Humanos , Pais , Análise de Regressão
9.
Acta Cir Bras ; 33(9): 736-743, 2018 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-30328905

RESUMO

PURPOSE: To evaluate osteocalcin gene and protein expression in vitro and in an in vivo model of ostectomy. METHODS: Twenty Wistar rats were assigned into two groups A (n=10, laser) and B (n=10, control). Ostectomy was performed in the femur diaphysis; the twenty fragments removed, composed in vitro groups named as in vivo (A and B) and cultivated in CO2 atmosphere for thirteen days. Low-level laser irradiation was performed in groups A (in vivo and in vitro) by an GaAlAs device (λ=808 nm, dose of 2J/cm2, power of 200mW, power density of 0.2W/cm2, total energy of 1.25J, spot diameter of 0.02mm) for 5 seconds, at one point, daily. It was performed immunocytochemistry assays in vivo and in vitro groups. In vitro groups were also submitted to RNA extraction, cDNA synthesis and gene expression by quantitative PCR. Statistical analysis was realized with p<0.05. RESULTS: Immunocytochemistry scores showed no significant differences between control and laser groups either in vivo and in vitro. Gene expression also showed no statistical differences. CONCLUSION: Low-level laser irradiation did not alter osteocalcin protein and gene expression in vivo and in vitro in the studied period but it may have been expressed them in an earlier period.


Assuntos
Fêmur/efeitos da radiação , Expressão Gênica/efeitos da radiação , Osteocalcina/efeitos da radiação , Animais , Fêmur/metabolismo , Fêmur/cirurgia , Imuno-Histoquímica , Terapia com Luz de Baixa Intensidade , Masculino , Modelos Animais , Osteocalcina/genética , Osteocalcina/metabolismo , Osteotomia , Ratos , Ratos Wistar
10.
Rev Soc Bras Med Trop ; 51(4): 528-532, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-30133639

RESUMO

INTRODUCTION: Molecular techniques for the detection of pathogens have been shown to be effective diagnostic tools with high sensitivity and short turnaround times. METHODS: This study compared five Staphylococcus aureus DNA extraction methods for detection by the polymerase chain reaction. RESULTS: The concentration and purity of the extracted DNA showed that the methods did not yield DNA of significant quality. However, most protocols yielded 100% positivity, even with low DNA concentrations. CONCLUSIONS: Although one protocol seemed more efficient than the others, PCR was sensitive enough to allow for detection of S. aureus with all the protocols.


Assuntos
Técnicas Bacteriológicas/métodos , DNA Bacteriano/isolamento & purificação , Reação em Cadeia da Polimerase/métodos , Staphylococcus aureus/genética , DNA Bacteriano/genética , Sensibilidade e Especificidade
11.
Cells ; 7(11)2018 Nov 02.
Artigo em Inglês | MEDLINE | ID: mdl-30400186

RESUMO

This study assessed whether telomere length is related to chronic conditions, cardiovascular risk factors, and inflammation in women aged 65 to 74 from Northeast Brazil. Participants were selected from two sources, a representative sample of the International Mobility in Aging Study (n = 57) and a convenience sample (n = 49) recruited at senior centers. Leukocyte telomere length was measured by quantitative polymerase chain reaction from blood samples in 83 women. Natural log-transformed telomere/single copy gene ratio was used as the dependent variable in the analysis. Blood analyses included inflammatory markers (high-sensitivity C-reactive protein and interleukin-6), total, low-density lipoprotein and high-density lipoprotein cholesterol, triglycerides, glucose and glycosylated hemoglobin. Self-rated health, chronic conditions, cardiovascular risk factors and inflammatory markers were not associated with telomere length. No significant independent association was found between telomere length and anthropometric measures or blood markers, even after adjusting for age, education and adverse childhood events among these older women in Northeast Brazil. Our results did not confirm the hypothesis that chronic conditions, cardiovascular risk factors or inflammation are associated with shorter telomere length in these women who have exceptional survival relative to the life expectancy of their birth cohort.

12.
PLoS One ; 12(7): e0180841, 2017.
Artigo em Inglês | MEDLINE | ID: mdl-28700679

RESUMO

Immunosenescence is an age-related reduction of immune system activity that can be associated with frailty. This study aimed to compare cytomegalovirus (CMV) and Epstein-Barr virus (EBV) reactivations (based on viremias) between young and elderly women who had a chronic CMV and/or EBV infection (i.e., an IgG+ serostatus) without an acute infection (i.e., an IgM- serostatus), and among the elderly group categorized according to frailty status. DNA was extracted from plasma using standard protocols and serostatus was determined by enzyme-linked immunosorbent assay. Quantitative real-time polymerase chain reaction analyses for CMV and EBV were carried out and viral loads were determined. Among elderly women (n = 71), 59% were positive for CMV, in contrast to only 8% of young women (n = 73). Elderly women classified as frail, pre-frail, and non-frail presented 82%, 56%, and 48% positivity for CMV, respectively. Frequency and viral load were significantly higher in the elderly group vs. the young group (p < 0.0001 and p = 0.01, respectively) and in elderly with frailty vs. those without frailty (p = 0.007 and p = 0.03, respectively). The frequency of CMV reactivation presented odds ratios of 11.77 for aging and 6.13 for frailty, and relative risks of 5.39 for aging and 1.93 for frailty. EBV was detected in 30% of the elderly women and 15% of the young women (p = 0.04); however, the viral load did not significantly differ between the two age groups. The frequency of EBV reactivation presented odds ratios of 2.36 for aging and 2.90 for frailty, and relative risks of 1.96 for aging and 2.12 for frailty. However, no difference in EBV viral load among the frailty status subgroups was found. In conclusion, the frequency of CMV reactivation was associated with aging and ongoing frailty, whereas the frequency of EBV reactivation was associated only with aging.


Assuntos
Envelhecimento/fisiologia , Citomegalovirus/fisiologia , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Envelhecimento/genética , Citomegalovirus/genética , Ensaio de Imunoadsorção Enzimática , Infecções por Vírus Epstein-Barr/virologia , Feminino , Idoso Fragilizado/estatística & dados numéricos , Humanos , Pessoa de Meia-Idade , Plasmídeos/genética , Reação em Cadeia da Polimerase em Tempo Real , Adulto Jovem
13.
Rev. Soc. Bras. Med. Trop ; 53: e20190181, 2020. graf
Artigo em Inglês | LILACS | ID: biblio-1092205

RESUMO

Abstract INTRODUCTION: Human herpesvirus (HHV)-7 establishes a latent infection during the lifetime of the host and can reactivate after the primary infection, leading to lytic replication in immunosuppressed patients. METHODS: This study aimed to develop an enzyme-linked immunosorbent assay (ELISA) to identify HHV-7 serum antibodies and compare its performance with that of an indirect immunofluorescence assay (IFA). RESULTS: Serum samples (n=102) were tested by IgG-IFA and by ELISA. IFA and ELISA showed IgG-positive results in 77 and 73 samples, respectively. Qualitative concordance of 96% was demonstrated between the two techniques. CONCLUSIONS: ELISA may be useful to diagnose HHV-7 infection.


Assuntos
Humanos , Imunoglobulina G/sangue , Herpesvirus Humano 7/imunologia , Infecções por Roseolovirus/diagnóstico , Anticorpos Antivirais/sangue , Ensaio de Imunoadsorção Enzimática , Curva ROC , Sensibilidade e Especificidade , Técnica Indireta de Fluorescência para Anticorpo
14.
Rev. Soc. Bras. Med. Trop ; 51(4): 528-532, July-Aug. 2018. tab, graf
Artigo em Inglês | LILACS | ID: biblio-1041475

RESUMO

Abstract INTRODUCTION Molecular techniques for the detection of pathogens have been shown to be effective diagnostic tools with high sensitivity and short turnaround times. METHODS This study compared five Staphylococcus aureus DNA extraction methods for detection by the polymerase chain reaction. RESULTS: The concentration and purity of the extracted DNA showed that the methods did not yield DNA of significant quality. However, most protocols yielded 100% positivity, even with low DNA concentrations. CONCLUSIONS: Although one protocol seemed more efficient than the others, PCR was sensitive enough to allow for detection of S. aureus with all the protocols.


Assuntos
Staphylococcus aureus/genética , DNA Bacteriano/isolamento & purificação , Reação em Cadeia da Polimerase/métodos , Técnicas Bacteriológicas/métodos , DNA Bacteriano/genética , Sensibilidade e Especificidade
15.
Acta cir. bras ; 33(9): 736-743, Sept. 2018. tab, graf
Artigo em Inglês | LILACS | ID: biblio-973502

RESUMO

Abstract Purpose: To evaluate osteocalcin gene and protein expression in vitro and in an in vivo model of ostectomy. Methods: Twenty Wistar rats were assigned into two groups A (n=10, laser) and B (n=10, control). Ostectomy was performed in the femur diaphysis; the twenty fragments removed, composed in vitro groups named as in vivo (A and B) and cultivated in CO2 atmosphere for thirteen days. Low-level laser irradiation was performed in groups A (in vivo and in vitro) by an GaAlAs device (λ=808 nm, dose of 2J/cm2, power of 200mW, power density of 0.2W/cm2, total energy of 1.25J, spot diameter of 0.02mm) for 5 seconds, at one point, daily. It was performed immunocytochemistry assays in vivo and in vitro groups. In vitro groups were also submitted to RNA extraction, cDNA synthesis and gene expression by quantitative PCR. Statistical analysis was realized with p<0.05. Results: Immunocytochemistry scores showed no significant differences between control and laser groups either in vivo and in vitro. Gene expression also showed no statistical differences. Conclusion: Low-level laser irradiation did not alter osteocalcin protein and gene expression in vivo and in vitro in the studied period but it may have been expressed them in an earlier period.


Assuntos
Animais , Masculino , Ratos , Expressão Gênica/efeitos da radiação , Osteocalcina/efeitos da radiação , Fêmur/efeitos da radiação , Osteotomia , Imuno-Histoquímica , Osteocalcina/genética , Osteocalcina/metabolismo , Ratos Wistar , Modelos Animais , Terapia com Luz de Baixa Intensidade , Fêmur/cirurgia , Fêmur/metabolismo
16.
Clinics (Sao Paulo) ; 66(6): 949-53, 2011.
Artigo em Inglês | MEDLINE | ID: mdl-21808857

RESUMO

OBJECTIVE: The aim of this study was to simultaneously monitoring cytomegalovirus and human herpesvirus 6 active infections using nested-polymerase chain reaction and, together with clinical findings, follow the clinical status of patients undergoing liver transplant. INTRODUCTION: The human ß-herpesviruses, including cytomegalovirus and human herpesvirus 6, are ubiquitous among human populations. Active infections of human herpesvirus 6 and cytomegalovirus are common after liver transplantation, possibly induced and facilitated by allograft rejection and immunosuppressive therapy. Both viruses affect the success of the transplant procedure. METHODS: Thirty patients submitted to liver transplant at the Liver Transplant Unit, at the Gastro Center, State University of Campinas, SP, Brazil, were studied prospectively from six months to one year, nested-polymerase chain reaction for cytomegalovirus and human herpesvirus 6 DNA detections. Two or more consecutive positive nested-polymerase chain reaction were considered indicative of active infection. RESULTS: Active infection by cytomegalovirus was detected in 13/30 (43.3%) patients, median time to first cytomegalovirus detection was 29 days after transplantation (range: 0-99 days). Active infection by human herpesvirus 6 was detected in 12/30 (40%) patients, median time to first human herpesvirus 6 detection was 23.5 days after transplantation (range: 0-273 days). The time-related appearance of each virus was not statistically different (p = 0.49). Rejection of the transplanted liver was observed in 16.7% (5/30) of the patients. The present analysis showed that human herpesvirus 6 and/or cytomegalovirus active infections were frequent in liver transplant recipients at our center. CONCLUSIONS: Few patients remain free of betaherpesviruses after liver transplantation. Most patients presenting active infection with more than one virus were infected sequentially and not concurrently. Nested-polymerase chain reaction can be considered of limited value for clinically monitoring cytomegalovirus and human herpesvirus 6.


Assuntos
Infecções por Citomegalovirus/diagnóstico , Citomegalovirus/isolamento & purificação , Herpesvirus Humano 6/isolamento & purificação , Transplante de Fígado/efeitos adversos , Infecções por Roseolovirus/diagnóstico , Citomegalovirus/genética , DNA Viral/análise , DNA Viral/genética , Seguimentos , Rejeição de Enxerto/virologia , Herpesvirus Humano 6/genética , Humanos , Transplante de Fígado/imunologia , Reação em Cadeia da Polimerase , Complicações Pós-Operatórias/diagnóstico , Complicações Pós-Operatórias/virologia , Estudos Prospectivos , Estatísticas não Paramétricas , Fatores de Tempo
17.
Rev Soc Bras Med Trop ; 41(6): 556-9, 2008.
Artigo em Inglês | MEDLINE | ID: mdl-19142431

RESUMO

Diagnosis of human herpesvirus-7 active infection in transplant patients has proved difficult, because this virus is ubiquitous and can cause persistent infections in the host. The significance of viral DNA detected in leukocytes by PCR is unclear and cross-reaction in serological tests may occur. This study aimed to evaluate nested-PCR to detect human herpesvirus-7 active infection in liver transplant recipients compared to healthy individuals. human herpesvirus-7 nested-PCR was performed on leukocytes and sera of 53 healthy volunteers and sera of 29 liver transplant recipients. In healthy volunteers, human herpesvirus-7 was detected in 28.3% of leukocytes and 0% of serum. human herpesvirus-7 was detected in sera of 48.2% of the liver transplant recipients. Nested-PCR on DNA extracted from leukocytes detected latent infection and the study suggests that nested-PCR performed on serum could be useful to detect human herpesvirus-7 active infection in liver transplant recipients.


Assuntos
DNA Viral/sangue , Herpesvirus Humano 7/isolamento & purificação , Transplante de Fígado , Reação em Cadeia da Polimerase/métodos , Infecções por Roseolovirus/diagnóstico , Adolescente , Adulto , Idoso , Estudos de Casos e Controles , Feminino , Herpesvirus Humano 7/genética , Humanos , Leucócitos Mononucleares/virologia , Masculino , Pessoa de Meia-Idade , Adulto Jovem
18.
Clinics ; 66(6): 949-953, 2011. ilus, tab
Artigo em Inglês | LILACS | ID: lil-594360

RESUMO

OBJECTIVE: The aim of this study was to simultaneously monitoring cytomegalovirus and human herpesvirus 6 active infections using nested-polymerase chain reaction and, together with clinical findings, follow the clinical status of patients undergoing liver transplant. INTRODUCTION: The human β-herpesviruses, including cytomegalovirus and human herpesvirus 6, are ubiquitous among human populations. Active infections of human herpesvirus 6 and cytomegalovirus are common after liver transplantation, possibly induced and facilitated by allograft rejection and immunosuppressive therapy. Both viruses affect the success of the transplant procedure. METHODS: Thirty patients submitted to liver transplant at the Liver Transplant Unit, at the Gastro Center, State University of Campinas, SP, Brazil, were studied prospectively from six months to one year, nested-polymerase chain reaction for cytomegalovirus and human herpesvirus 6 DNA detections. Two or more consecutive positive nested-polymerase chain reaction were considered indicative of active infection. RESULTS: Active infection by cytomegalovirus was detected in 13/30 (43.3 percent) patients, median time to first cytomegalovirus detection was 29 days after transplantation (range: 0-99 days). Active infection by human herpesvirus 6 was detected in 12/30 (40 percent) patients, median time to first human herpesvirus 6 detection was 23.5 days after transplantation (range: 0-273 days). The time-related appearance of each virus was not statistically different (p = 0.49). Rejection of the transplanted liver was observed in 16.7 percent (5/30) of the patients. The present analysis showed that human herpesvirus 6 and/or cytomegalovirus active infections were frequent in liver transplant recipients at our center. CONCLUSIONS: Few patients remain free of betaherpesviruses after liver transplantation. Most patients presenting active infection with more than one virus were infected sequentially and not concurrently. Nested-polymerase chain reaction can be considered of limited value for clinically monitoring cytomegalovirus and human herpesvirus 6.


Assuntos
Humanos , Infecções por Citomegalovirus/diagnóstico , Citomegalovirus/isolamento & purificação , /isolamento & purificação , Transplante de Fígado/efeitos adversos , Infecções por Roseolovirus/diagnóstico , Citomegalovirus/genética , DNA Viral/análise , DNA Viral/genética , Seguimentos , Rejeição de Enxerto/virologia , /genética , Transplante de Fígado/imunologia , Reação em Cadeia da Polimerase , Estudos Prospectivos , Complicações Pós-Operatórias/diagnóstico , Complicações Pós-Operatórias/virologia , Estatísticas não Paramétricas , Fatores de Tempo
19.
Rev. Soc. Bras. Med. Trop ; 41(6): 556-559, Nov.-Dec. 2008. ilus, graf
Artigo em Inglês | LILACS | ID: lil-502033

RESUMO

Diagnosis of human herpesvirus-7 active infection in transplant patients has proved difficult, because this virus is ubiquitous and can cause persistent infections in the host. The significance of viral DNA detected in leukocytes by PCR is unclear and cross-reaction in serological tests may occur. This study aimed to evaluate nested-PCR to detect human herpesvirus-7 active infection in liver transplant recipients compared to healthy individuals. human herpesvirus-7 nested-PCR was performed on leukocytes and sera of 53 healthy volunteers and sera of 29 liver transplant recipients. In healthy volunteers, human herpesvirus-7 was detected in 28.3 percent of leukocytes and 0 percent of serum. human herpesvirus-7 was detected in sera of 48.2 percent of the liver transplant recipients. Nested-PCR on DNA extracted from leukocytes detected latent infection and the study suggests that nested-PCR performed on serum could be useful to detect human herpesvirus-7 active infection in liver transplant recipients.


Diagnóstico da infecção ativa pelo herpesvirus humano-7 é difícil devido ao fato deste vírus ser ubíquo e poder causar infecção persistente no hospedeiro. O significado da detecção do DNA viral por reação em cadeia da polimerase não é claro e, reações cruzadas podem ocorrer em testes sorológicos. O objetivo deste estudo foi avaliar a nested-PCR para detectar infecção ativa pelo herpesvirus-7 em receptores hepáticos comparando com indivíduos sadios. Nested-PCR para herpesvirus-7 foi realizado em leucócitos e soro de 53 voluntários sadios e em soro de 29 receptores hepáticos. Nos voluntários sadios, herpesvirus-7 foi detectado em 28,3 por cento de leucócitos e 0 por cento de soro. herpesvirus-7 foi detectado em soro de 48,2 por cento de receptores hepáticos. Nested-PCR em DNA extraído de leucócitos detectou infecção latente e o estudo sugere que nested-PCR realizada em soro poderia ser útil para detectar infecção ativa por herpesvirus-7 em receptores de fígado.


Assuntos
Adolescente , Adulto , Idoso , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Adulto Jovem , DNA Viral/sangue , /isolamento & purificação , Transplante de Fígado , Reação em Cadeia da Polimerase/métodos , Infecções por Roseolovirus/diagnóstico , Estudos de Casos e Controles , /genética , Leucócitos Mononucleares/virologia , Adulto Jovem
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