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1.
Article in Chinese | WPRIM | ID: wpr-936145

ABSTRACT

OBJECTIVE@#To investigate the consistency of cytomegalovirus deoxyribo nucleic acid (CMV-DNA) and immunoglobulin M (IgM) antibody detections in patients with different clinical characteristics and their guiding value for clinical practice.@*METHODS@#From December 2014 to November 2019, a total of 507 patients who were detected with both CMV-IgM and CMV-DNA were collected in Peking University International Hospital. Their general information, such as gender, age and clinical data, including the patient's diagnosis, medication, and outcome were also collected. The groups were stratified according to whether CMV-DNA was negative or positive, CMV-IgM was negative or positive, age, gender, and whether they received immunosuppressive therapy or not. The Pearson Chi-square test or Fisher's exact test was used for comparison of the rates between the groups. P < 0.05 means the difference is statisti-cally significant.@*RESULTS@#Of the 507 patients submitted for examination, 55 (10.85%) were positive for CMV-DNA, 74 (14.60%) were positive for CMV-IgM, and 20 (3.94%) were positive for both CMV-DNA and CMV-IgM. Of the 55 patients with CMV-DNA positive, 37 were male, accounting for 67.27%. In addition, 25 patients were older than 60 years, accounting for 45.45% and 33 patients received immunosuppressive therapy, accounting for 60%. The rates were higher than that of CMV-DNA negative group, 47.35% (P=0.005), 68.14% (P=0.043), 46.02% (P=0.050), respectively. Of the patients with both CMV-DNA and IgM positive, 45% received immunosuppressive threapy, which was lower than that of CMV-DNA positive but IgM negative patients (68.57%, P=0.086), and also lower than CMV-DNA negative but IgM positive patients (68.52%, P=0.064). In the patients with both CMV-DNA and IgM positive, 91.67% showed remission after receiving ganciclovir, whereas in the patients with CMV-DNA positive but IgM negative, the rate was only 60% (P=0.067).@*CONCLUSION@#CMV-IgM antibody detection is affected by age, gender, and immune status. It is not recommended to use CMV-IgM alone to determine CMV infection in patients with immunosuppressive status and those older than 60 years. CMV-DNA and CMV-IgM combined detection may help to predict patients' immune status and outcomes of antiviral therapy.


Subject(s)
Antibodies, Viral , Cytomegalovirus/genetics , Cytomegalovirus Infections/drug therapy , DNA , Female , Humans , Immunoglobulin M , Immunosuppressive Agents/therapeutic use , Male , Nucleic Acids
2.
Article in English | WPRIM | ID: wpr-887743

ABSTRACT

Epstein-Barr virus (EBV) and cytomegalovirus (CMV), two of the most prevalent human herpesviruses, cause a wide spectrum of diseases and symptoms and are associated with serious health problem. In this study, we developed an internal control reference recombinase-aided amplification (ICR-RAA) assay for the rapid detection of EBV and CMV within 30 min. The assay had a sensitivity of 5 and 1 copies/test for EBV and CMV, respectively, with no cross reaction with other pathogens. In comparison with those of the commercial quantitative polymerase chain reaction (qPCR), the sensitivity of the EBV and CMV ICR-RAAs using extracted DNA was 93.33% and 84.84%, respectively; the specificity was 98.75% and 100.00%, respectively; and the Kappa values were 0.930 and 0.892 (


Subject(s)
Adolescent , Adult , Child , Child, Preschool , Cytomegalovirus/genetics , Cytomegalovirus Infections/virology , DNA, Viral/analysis , Epstein-Barr Virus Infections/virology , Female , Herpesvirus 4, Human/genetics , Humans , Infant , Infant, Newborn , Male , Middle Aged , Nucleic Acid Amplification Techniques , Recombinases/genetics , Young Adult
3.
Clinics ; 75: e1498, 2020. tab, graf
Article in English | LILACS | ID: biblio-1055886

ABSTRACT

OBJECTIVE: To evaluate the role of intraocular fluid analysis as a diagnostic aid for uveitis. METHODS: Twenty-eight samples (27 patients including 3 HIV-infected patients) with active (n=24) or non-active (n=4) uveitis were submitted to aqueous (AH; n=12) or vitreous humor (VH) analysis (n=16). All samples were analyzed by quantitative PCR for herpes simplex virus (HSV), varicella zoster virus (VZV), cytomegalovirus (CMV), Epstein-Barr virus (EBV) and Toxoplasma gondii. RESULTS: The positivity of the PCR in AH was 41.7% (5/12), with 50% (2/4) in immunocompetent and 67% (2/3) in HIV+ patients. The positivity of the PCR in VH was 31.2% (5/16), with 13% (1/8) in immunocompetent and 50% (4/8) in immunosuppressed HIV negative patients. The analysis was a determinant in the diagnostic definition in 58% of HA and 50% of VH. CONCLUSION: Even in posterior uveitis, initial AH analysis may be helpful. A careful formulation of possible clinical diagnosis seems to increase the chance of intraocular sample analysis being meaningful.


Subject(s)
Humans , Aqueous Humor/microbiology , Aqueous Humor/parasitology , Aqueous Humor/virology , Uveitis/diagnosis , Vitreous Body/microbiology , Vitreous Body/parasitology , Toxoplasma , Uveitis/microbiology , Uveitis/parasitology , Uveitis/virology , Vitreous Body/virology , DNA, Viral/analysis , Polymerase Chain Reaction , HIV-1 , Immunocompromised Host , Simplexvirus/genetics , Simplexvirus/immunology , Herpesvirus 4, Human , Herpesvirus 3, Human/genetics , Herpesvirus 3, Human/immunology , Cytomegalovirus/genetics , Cytomegalovirus/immunology , Immunocompetence
4.
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
5.
Rev. Soc. Bras. Med. Trop ; 51(2): 141-145, Mar.-Apr. 2018. tab, graf
Article in English | LILACS | ID: biblio-897064

ABSTRACT

Abstract INTRODUCTION: Human cytomegalovirus is one of the causes of opportunist infections in immunocompromised patients, and is triggered by factors such as state of viral latency, weakened immune responses, and development of antiviral resistance to ganciclovir, the only drug offered by the public health system in Brazil to treat the infection. The goal of this study was to identify mutations that may be associated with antiviral resistance in immunocompromised patients. METHODS: Molecular analysis was performed in 82 blood samples and subjected to genomic DNA extraction by a silica-based method. Three sequences of the HCMV UL97 gene, which encodes a phosphotransferase protein required for activation of ganciclovir, were amplified by polymerase chain reaction. Pyrosequencing methods were applied to one external 2096-bp segment DNA and two internal sequences between nucleotides 1087 to 1828 to detect mutations in this gene. RESULTS: Approximately 10% of sequences contained mutations between nucleotides 377 and 594, in conserved regions of the UL97 gene, leading to amino acid changes. Eleven coding mutations were identified, including changes leading to amino acid substitutions, E596K and S604F, which were observed in 100% of samples and are described for the first time in Brazil. In addition, one mutation (A594V) that is associated with ganciclovir resistance was detected in a kidney transplant patient. CONCLUSIONS: Further studies to detect mutations associated with HCMV resistance to antiviral drugs are required to demonstrate the need to increase the variety and availability of drugs used to treat viral infections in the public health care system in Brazil.


Subject(s)
Humans , Antiviral Agents/therapeutic use , Phosphotransferases/genetics , Immunocompromised Host , Cytomegalovirus Infections/drug therapy , Cytomegalovirus/enzymology , Drug Resistance, Viral/genetics , Mutation/genetics , Antiviral Agents/pharmacology , Case-Control Studies , Polymerase Chain Reaction , Cross-Sectional Studies , Cytomegalovirus/drug effects , Cytomegalovirus/genetics , Drug Resistance, Viral/drug effects , Genotype
6.
Article in English | WPRIM | ID: wpr-8650

ABSTRACT

BACKGROUND: Cytomegalovirus (CMV) and Epstein-Barr virus (EBV) are increasingly important in immunocompromised patients. Nucleic acid extraction methods could affect the results of viral nucleic acid amplification tests. We compared two automated nucleic acid extraction systems for detecting CMV and EBV using real-time PCR assays. METHODS: One hundred and fifty-three whole blood (WB) samples were tested for CMV detection, and 117 WB samples were tested for EBV detection. Viral nucleic acid was extracted in parallel by using QIAsymphony RGQ and QIAcube (Qiagen GmbH, Germany), and real-time PCR assays for CMV and EBV were performed with a Rotor-Gene Q real-time PCR cycler (Qiagen). Detection rates for CMV and EBV were compared, and agreements between the two systems were analyzed. RESULTS: The detection rate of CMV and EBV differed significantly between the QIAsymphony RGQ and QIAcube systems (CMV, 59.5% [91/153] vs 43.8% [67/153], P=0.0005; EBV, 59.0% [69/117] vs 42.7% [50/117], P=0.0008). The two systems showed moderate agreement for CMV and EBV detection (kappa=0.43 and 0.52, respectively). QIAsymphony RGQ showed a negligible correlation with QIAcube for quantitative EBV detection. QIAcube exhibited EBV PCR inhibition in 23.9% (28/117) of samples. CONCLUSIONS: Automated nucleic acid extraction systems have different performances and significantly affect the detection of viral pathogens. The QIAsymphony RGQ system appears to be superior to the QIAcube system for detecting CMV and EBV. A suitable sample preparation system should be considered for optimized nucleic acid amplification in clinical laboratories.


Subject(s)
Automation , Cytomegalovirus/genetics , Cytomegalovirus Infections/diagnosis , DNA, Viral/blood , Herpesvirus 4, Human/genetics , Humans , Reagent Kits, Diagnostic , Real-Time Polymerase Chain Reaction
7.
Plos one ; 11(7)2016.
Article in English | LILACS, SES-SP, SESSP-IALPROD, SES-SP, SESSP-IALACERVO | ID: biblio-1024785

ABSTRACT

Gliomas are the most common type of primary brain tumors. The most aggressive type, Glioblastoma multiforme (GBM), is one of the deadliest human diseases, with an average survival at diagnosis of about 1 year. Previous evidence suggests a link between human cytomegalovirus (HCMV) and gliomas. HCMV has been shown to be present in these tumors and several viral proteins can have oncogenic properties in glioma cells. Here we have investigated the presence of HCMV DNA, RNA and proteins in fifty-two gliomas of different grades of malignancy. The UL83 viral region, the early beta 2.7 RNA and viral protein were detected in 73%, 36% and 57% by qPCR, ISH and IHC, respectively. Positivity of the viral targets and viral load was independent of tumor type or grade suggesting no correlation between viral presence and tumor progression. Our results demonstrate high prevalence of the virus in gliomas from Brazilian patients, contributing to a better understanding of the association between HCMV infection and gliomas worldwide and supporting further investigations of the virus oncomodulatory properties.


Subject(s)
Viral Proteins/genetics , Brazil , Humans , RNA, Viral , Immunohistochemistry , In Situ Hybridization , Cytomegalovirus Infections/complications , Viral Load , Cytomegalovirus/genetics , Glioma
8.
Article in English | WPRIM | ID: wpr-200494

ABSTRACT

Standardized cytomegalovirus (CMV) DNA quantification is important for managing CMV disease. We evaluated the performance of the Real-Q CMV Quantification Kit (Real-Q assay; BioSewoom, Korea) using whole blood (WB), with nucleic acid extraction using MagNA Pure 96 (Roche Diagnostics, Germany). Real-time PCR was performed on two platforms: the 7500 Fast real-time PCR (7500 Fast; Applied Biosystems, USA) and CFX96 real-time PCR detection (CFX96; Bio-Rad, USA) systems. The WHO international standard, diluted with CMV-negative WB, was used to validate the analytical performance. We used 90 WB clinical samples for comparison with the artus CMV RG PCR kit (artus assay; Qiagen, Germany). Limits of detections (LODs) in 7500 Fast and CFX96 were 367 and 479 IU/mL, respectively. The assay was linear from the LOD to 10(6) IU/mL (R2 ≥0.9886). The conversion factors from copies to IU in 7500 Fast and CFX96 were 0.95 and 1.06, respectively. Compared with the artus assay, for values 1,000 copies/mL, 73.3% and 80.6% of samples in 7500 Fast and CFX96, respectively, had <0.5 log10 copies/mL. The Real-Q assay is useful for quantifying CMV in WB with the two real-time PCR platforms.


Subject(s)
Cytomegalovirus/genetics , Cytomegalovirus Infections/diagnosis , DNA, Viral/blood , Humans , Limit of Detection , Reagent Kits, Diagnostic , Real-Time Polymerase Chain Reaction
9.
Braz. j. infect. dis ; 19(4): 403-409, July-Aug. 2015. tab, ilus
Article in English | LILACS | ID: lil-759282

ABSTRACT

Human cytomegalovirus is a ubiquitous pathogen that infects the majority of the world's population. After long period of time co-evolving with human being, this pathogen has developed several strategies to evade host immune surveillance. One of the major trick is encoding homologous to those of the host organism or stealing host cellular genes that have significant functions in immune system. To date, we have found several viral immune analogous which include G protein coupled receptor, class I major histocompatibility complex and chemokine. Chemokine is a small group of molecules which is defined by the presence of four cysteines in highly conserved region. The four kinds of chemokines (C, CC, CXC, and CX3C) are classified based on the arrangement of 1 or 2 N-terminal cysteine residues. UL128 protein is one of the analogous that encoded by human cytomegalovirus that has similar amino acid sequences to the human CC chemokine. It has been proved to be one of the essential particles that involved in human cytomegalovirus entry into epithelial/endothelial cells as well as macrophages. It is also the target of potent neutralizing antibodies in human cytomegalovirus-seropositive individuals. We had demonstrated the chemotactic trait of UL128 protein in our previous study. Recombinant UL128 in vitrohas the ability to attract monocytes to the infection region and enhances peripheral blood mononuclear cell proliferation by activating the MAPK/ERK signaling pathway. However, the way that this viral encoded chemokine interacting with peripheral blood mononuclear cells and the detailed mechanism that involving the virus entry into host cells keeps unknown. Here we performed in vitroinvestigation into the effects of UL128 protein on peripheral blood mononuclear cell's activation and receptor binding, which may help us further understand the immunomodulatory function of UL128 protein as well as human cytomegalovirus diffusion mechanism.


Subject(s)
Humans , Chemokines, CC , Cytomegalovirus , Gene Expression Regulation, Viral/genetics , Leukocytes, Mononuclear/virology , Membrane Glycoproteins/immunology , Viral Envelope Proteins/immunology , Cells, Cultured , Cross-Linking Reagents , Chemokines, CC/genetics , Chemokines, CC/immunology , Cytomegalovirus/genetics , Cytomegalovirus/immunology , Receptors, Chemokine/genetics , Recombinant Proteins/immunology
10.
Article in English | WPRIM | ID: wpr-34568

ABSTRACT

BACKGROUND: Quantitation of cytomegalovirus (CMV) DNA using real-time PCR has been utilized for monitoring CMV infection. However, the CMV antigenemia assay is still the 'gold standard' assay. There are only a few studies in Korea that compared the efficacy of use of real-time PCR for quantitation of CMV DNA in whole blood with the antigenemia assay, and most of these studies have been limited to transplant recipients. METHOD: 479 whole blood samples from 79 patients, falling under different disease groups, were tested by real-time CMV DNA PCR using the Q-CMV real-time complete kit (Nanogen Advanced Diagnostic S.r.L., Italy) and CMV antigenemia assay (CINA Kit, ArgeneBiosoft, France), and the results were compared. Repeatedly tested patients were selected and their charts were reviewed for ganciclovir therapy. RESULTS: The concordance rate of the two assays was 86.4% (Cohen's kappa coefficient value=0.659). Quantitative correlation between the two assays was a moderate (r=0.5504, P<0.0001). Among 20 patients tested repeatedly with the two assays, 13 patients were transplant recipients and treated with ganciclovir. Before treatment, CMV was detected earlier by real-time CMV DNA PCR than the antigenemia assay, with a median difference of 8 days. After treatment, the antigenemia assay achieved negative results earlier than real-time CMV DNA PCR with a median difference of 10.5 days. CONCLUSIONS: Q-CMV real-time complete kit is a useful tool for early detection of CMV infection in whole blood samples in transplant recipients.


Subject(s)
Antiviral Agents/therapeutic use , Cytomegalovirus/genetics , Cytomegalovirus Infections/drug therapy , DNA, Viral/blood , Ganciclovir/therapeutic use , Humans , Immunoassay , Organ Transplantation , Phosphoproteins/genetics , Real-Time Polymerase Chain Reaction , Viral Matrix Proteins/genetics , Virology/methods
11.
Article in English | WPRIM | ID: wpr-19703

ABSTRACT

We report a case of cytomegalovirus (CMV) retinitis after intravitreal bevacizumab injection. A 61-year-old woman with diabetic macular edema developed dense vitritis and necrotizing retinitis 3 weeks after intravitreal bevacizumab injection. A diagnostic vitrectomy was performed. The undiluted vitreous sample acquired by vitrectomy was analyzed by polymerase chain reaction and culture. Polymerase chain reaction of the vitreous was positive for CMV DNA. Other laboratory results did not show evidence of other infectious retinitis and systemic immune dysfunction. Human immunodeficiency virus antibodies were also negative. After systemic administration of ganciclovir, retinitis has resolved and there has been no recurrence of retinitis during the follow-up period of 12 months. Ophthalmologists should be aware of potential risk for CMV retinitis after intravitreal bevacizumab injection.


Subject(s)
Angiogenesis Inhibitors/administration & dosage , Antibodies, Monoclonal, Humanized/administration & dosage , Cytomegalovirus/genetics , Cytomegalovirus Retinitis/diagnosis , DNA, Viral/analysis , Diagnosis, Differential , Female , Humans , Immunocompetence/drug effects , Intravitreal Injections , Macular Edema/diagnosis , Middle Aged , Polymerase Chain Reaction , Vascular Endothelial Growth Factor A/antagonists & inhibitors
12.
Article in Korean | WPRIM | ID: wpr-169731

ABSTRACT

Cytomegalovirus (CMV) infections are usually diagnosed in immunocompromised patients. A 74-year-old male without any significant medical history visited our center because of abdominal pain and diarrhea which began about a month ago. Abdominal computed tomography revealed segmental enhanced bowel wall thickening on jejunum and single-balloon enteroscopy showed multiple geographic shaped ulcerations covered with exudates on proximal jejunum. Biopsy samples taken during endoscopic examination demonstrated necrotic fibrinopurulent tissue debris and benign ulcer. Nested-PCR analysis of CMV DNA from jejunal tissue was positive. The patient was finally diagnosed with CMV jejunitis and was treated by intravenous ganciclovir for 14 days after which, abdominal pain and diarrhea improved. Our case shows that CMV jejunitis can occur in an immunocompetent adult as multiple jejunal ulcers which can be diagnosed using a single-balloon enteroscope.


Subject(s)
Aged , Antiviral Agents/therapeutic use , Cytomegalovirus/genetics , Cytomegalovirus Infections/complications , DNA, Viral/analysis , Endoscopy, Gastrointestinal , Enteritis/diagnosis , Ganciclovir/therapeutic use , Humans , Injections, Intravenous , Jejunal Diseases/diagnosis , Male , Polymerase Chain Reaction , Tomography, X-Ray Computed
13.
Mem. Inst. Oswaldo Cruz ; 107(7): 953-954, Nov. 2012. ilus, tab
Article in English | LILACS | ID: lil-656057

ABSTRACT

Members of the Herpesviridae family have been implicated in a number of tumours in humans. At least 75% of the human population has had contact with cytomegalovirus (HCMV). In this work, we screened 75 Brazilian glioma biopsies for the presence of HCMV DNA sequences. HCMV DNA was detected in 36% (27/75) of the biopsies. It is possible that HCMV could be a co-factor in the evolution of brain tumours.


Subject(s)
Adult , Child , Female , Humans , Male , Young Adult , Brain Neoplasms/virology , Cytomegalovirus Infections/complications , Cytomegalovirus/genetics , DNA, Viral/analysis , Glioma/virology , Biopsy , Cohort Studies , Cytomegalovirus Infections/diagnosis , Cytomegalovirus/immunology , Immediate-Early Proteins/analysis , Immediate-Early Proteins/immunology , Neoplasm Staging , Polymerase Chain Reaction , Prevalence
15.
Article in English | WPRIM | ID: wpr-102516

ABSTRACT

Hepatitis A virus (HAV) infections occur predominantly in children, and are usually self-limiting. However, 75-95% of the infections in adults are symptomatic (mostly with jaundice), with the illness symptoms usually persisting for a few weeks. Atypical manifestations include relapsing hepatitis, prolonged cholestasis, and complications involving renal injury. Drug rash with eosinophilia and systemic symptoms (DRESS) syndrome is a severe, drug-induced hypersensitivity reaction characterized by skin rash, fever, lymph-node enlargement, and internal organ involvement. We describe a 22-year-old male who presented with acute kidney injury and was diagnosed with prolonged cholestatic hepatitis A. The patient also developed DRESS syndrome due to antibiotic and/or antiviral treatment. To our knowledge, this is the first report of histopathologically confirmed DRESS syndrome due to antibiotic and/or antiviral treatment following HAV infection with cholestatic features and renal injury.


Subject(s)
Acute Kidney Injury/diagnosis , Anti-Bacterial Agents/adverse effects , Cefotaxime/adverse effects , Cholestasis/complications , Cytomegalovirus/genetics , Cytomegalovirus Infections/drug therapy , DNA, Viral/analysis , Eosinophilia/etiology , Exanthema/chemically induced , Ganciclovir/therapeutic use , Hepatitis A/complications , Humans , Hydrocortisone/therapeutic use , Immunoglobulins/therapeutic use , Male , Syndrome , Young Adult
16.
Rev. Soc. Bras. Med. Trop ; 44(5): 551-554, Sept.-Oct. 2011. ilus, tab
Article in English | LILACS | ID: lil-602919

ABSTRACT

INTRODUCTION: Human cytomegalovirus is an opportunistic betaherpesvirus that causes persistent and serious infections in immunodeficient patients. Recurrent infections occur due to the presence of the virus in a latent state in some cell types. It is possible to examine the virus using molecular methods to aid in the immunological diagnosis and to generate a molecular viral profile in immunodeficient patients. The objective of this study was to characterize cytomegalovirus genotypes and to generate the epidemiological and molecular viral profile in immunodeficient patients. METHODS: A total of 105 samples were collected from immunodeficient patients from the City of Belém, including newborns, hemodialysis patients, transplant recipients and HIV+ patients. An IgG and IgM antibody study was completed using ELISA, and enzymatic analysis by restriction fragment length polymorphism (RFLP) was performed to characterize viral genotypes. RESULTS: It was observed that 100 percent of the patients had IgG antibodies, 87 percent of which were IgG+/IgM-, consistent with a prior infection profile, 13 percent were IgG+/IgM+, suggestive of recent infection. The newborn group had the highest frequency (27 percent) of the IgG+/IgM+ profile. By RFLP analysis, only one genotype was observed, gB2, which corresponded to the standard AD169 strain. CONCLUSIONS: The presence of IgM antibodies in new borns indicates that HCMV continues to be an important cause of congenital infection. The low observed genotypic diversity could be attributed to the small sample size because newborns were excluded from the RFLP analysis. This study will be continued including samples from newborns to extend the knowledge of the general and molecular epidemiology of HCMV in immunodeficient patients.


INTRODUÇÃO: O citomegalovírus é um betaherpesvírus oportunista, causador de infecções persistentes e graves em pacientes imunodeficientes. As infecções recorrentes ocorrem devido à presença do vírus em estado de latência, em alguns tipos celulares, o que possibilita a pesquisa viral por métodos moleculares para auxiliar nos diagnósticos imunológicos, assim como traçar o perfil epidemiológico e molecular viral em pacientes imunodeficientes. O objetivo deste estudo foi caracterizar os genótipos de citomegalovírus e traçar o perfil epidemiológico e molecular viral em pacientes imunodeficientes. MÉTODOS: Um total de 105 amostras foi coletado de pacientes imunodeficientes da Cidade de Belém, incluindo recém-nascidos, hemodialisados, transplantados e pacientes HIV+. Foi realizada a pesquisa de anticorpos IgG e IgM pelo método ELISA e análise enzimática pelo método restriction fragment length polymorphism (RFLP) para caracterização dos genótipos virais. RESULTADOS: Foi observado que 100 por cento dos pacientes apresentavam anticorpos IgG, 87 por cento eram IgG+/IgM-perfil de infecção pregressa; e 13 por cento IgG+/ IgM+ sugestivo de infecção recente. O grupo dos recém-nascidos apresentou maior frequência (27 por cento) do perfil IgG+/IgM+. Na análise por RFLP, foi observado um único genótipo, o gB2, que corresponde ao padrão genotípico da cepa AD169. CONCLUSÕES: A presença de anticorpos IgM nos recém-nascidos indica que o vírus CMV continua sendo causa importante de infecção congênita; a baixa diversidade genotípica pode ser atribuída ao tamanho amostral devido a exclusão dos recém-nascidos na análise por RFLP. Esse estudo será continuado incluindo amostras de recém-nascidos a fim de contribuir para um amplo conhecimento da epidemiologia geral e molecular do citomegalovírus em pacientes imunodeficientes da Cidade de Belém.


Subject(s)
Adult , Humans , Infant, Newborn , Cytomegalovirus Infections/virology , Cytomegalovirus/genetics , Genome, Viral/genetics , HIV Infections/immunology , Immunocompromised Host/immunology , Kidney Transplantation/immunology , Brazil , Dialysis , Immunoglobulin G/blood , Immunoglobulin M/blood , Restriction Mapping/methods
17.
Braz. j. infect. dis ; 15(5): 484-485, Sept.-Oct. 2011. ilus
Article in English | LILACS | ID: lil-612710

ABSTRACT

Cytomegalovirus (CMV) infection is a common opportunistic systemic infection in immunocompromised patients, but skin involvement is rare. Herein, we report a 10 year-old girl from consanguineous parents who was referred to our center because of disseminated maculopapular rash. She had history of upper and lower respiratory tract infections. In immunological studies, increased serum IgE level and decreased responses to tetanus and diphtheria were detected. Polymerase chain reaction (PCR) examination of bronchoalveolar lavage and serum sample revealed the presence of CMV. Early diagnosis of cutaneous CMV and appropriate treatment are the key actions in management of patients with underlying immunodeficiencies to avoid further complications.


Subject(s)
Child , Female , Humans , Cytomegalovirus Infections/diagnosis , Cytomegalovirus/immunology , Immunocompromised Host/immunology , Immunoglobulin E/blood , Skin Diseases, Viral/diagnosis , Cytomegalovirus Infections/immunology , Cytomegalovirus/genetics , DNA, Viral/blood , DNA, Viral/immunology , Polymerase Chain Reaction , Skin Diseases, Viral/immunology
18.
Rev. Soc. Bras. Med. Trop ; 44(3): 286-289, May-June 2011. ilus, tab
Article in English | LILACS | ID: lil-593353

ABSTRACT

INTRODUCTION: Human cytomegalovirus (HCMV) is often reactive in latently infected immunosuppressed patients. Accordingly, HCMV remains one of the most common infections following solid organ and hemopoietic stem cell transplantations, resulting in significant morbidity, graft loss and occasional mortality. The early diagnosis of HCMV disease is important in immunosuppressed patients, since in these individuals, preemptive treatment is useful. The objective of this study was to compare the performance of the in-house qualitative polymerase chain reaction (PCR) and pp65 antigenemia to HCMV infection in immunosuppressed patients in the Hospital de Clínicas of Porto Alegre (HCPA). METHODS: A total of 216 blood samples collected between August 2006 and January 2007 were investigated. RESULTS: Among the samples analyzed, 81 (37.5 percent) were HCMV-positive by PCR, while 48 (22.2 percent) were positive for antigenemia. Considering antigenemia as the gold standard, sensitivity, specificity, positive predictive values and negative predictive values for PCR were 87.5 percent, 76.8 percent, 51.8 percent and 95.5 percent respectively. CONCLUSIONS: These results demonstrated that qualitative PCR has high sensitivity and negative predictive value (NPV). Consequently PCR is especially indicated for the initial diagnosis of HCMV infection. In the case of preemptive treatment strategy, identification of patients at high-risk for HCMV disease is fundamental and PCR can be useful tool.


INTRODUÇÃO: O citomegalovírus humano (HCMV), causador de infecção latente, reativa com frequência em pacientes imunossuprimidos. Portanto, o HCMV permanece uma das infecções mais comuns após transplantes de órgãos sólidos e de células hematopoiéticas resultando em significativa morbidade, perda do enxerto e ocasional mortalidade. Assim, o diagnóstico precoce para uma terapia preventiva é de grande importância. Este estudo visa comparar o desempenho dos métodos PCR qualitativo in-house e antigenemia pp65 para o diagnóstico de infecção por CMV em pacientes imunossuprimidos do Hospital de Clínicas de Porto Alegre. MÉTODOS: O estudo foi realizado em 216 amostras de sangue total (EDTA) coletadas de 85 pacientes, entre agosto de 2006 e janeiro de 2007. RESULTADOS: Dentre as 216 amostras analisadas, 81 (37,5 por cento) amostras apresentaram resultados positivos na PCR, enquanto 48 (22,2 por cento) apresentaram resultados positivos na antigenemia. A sensibilidade, especificidade, valor preditivo positivo e valor preditivo negativo para a PCR, considerando antigenemia como padrão foram 87,5 por cento, 76,8 por cento, 51,8 por cento e 95,5 por cento, respectivamente. CONCLUSÕES: Estes resultados demonstraram que a PCR tem alta sensibilidade e valor preditivo negativo. Consequentemente PCR é especialmente indicada para o diagnóstico inicial de infecção por HCMV. No caso da estratégia de terapia preventiva, a identificação de pacientes com alto risco para a doença por HCMV é fundamental e a PCR pode ser uma ferramenta útil.


Subject(s)
Humans , Antigens, Viral/analysis , Cytomegalovirus Infections/diagnosis , Cytomegalovirus/genetics , Cytomegalovirus/immunology , DNA, Viral/analysis , Immunocompromised Host/immunology , Cytomegalovirus Infections/immunology , Predictive Value of Tests , Phosphoproteins/immunology , Polymerase Chain Reaction/methods , Reproducibility of Results , Sensitivity and Specificity , Viral Matrix Proteins/immunology
19.
Rev. Inst. Med. Trop. Säo Paulo ; 53(2): 82-88, Mar.-Apr. 2011. tab
Article in English | LILACS | ID: lil-584138

ABSTRACT

BACKGROUND: Cytomegalovirus (CMV) remains an important pathogen to immunocompromised patients even in the era of HAART. The present study aimed at evaluating the influence of CMV viral load and its gB genotypes on AIDS patients' outcome. METHODS: Blood samples of 101 AIDS patients were collected and tested for HIV load, CD4 - cell count and opportunistic pathogens, including CMV. Semi-nested PCRs were run to detect CMV genome and in the positive samples, gB genotyping and CMV load were established using enzymatic restriction and real time PCR, respectively. All patients were clinically followed for four years. RESULTS: In thirty patients (31 percent) CMV was detected and all fatal cases (n = 5) occurred in this group of patients (p = 0.007), but only two patients had CMV disease (1.9 percent). However, viral load was not statistically associated with any analyzed parameter. The most frequently observed CMV genotype was gB2 (45.16 percent) followed by gB3 (35.48 percent). gB2 genotype was more frequently found in patients with CD4-cell counts under 200 cells/mm³ (p = 0.0017), and almost all fatal cases (80 percent) had gB2 genotype. CONCLUSIONS: Our study suggests that CMV and its polymorphisms in biologically relevant genes, such as the gB encoding ORF, may still influence the prognosis and outcome of AIDS patients. The gB2 genotype was associated to patient's bad outcome.


ANTECEDENTES: O citomegalovírus (CMV) permanece um importante patógeno para pacientes imunocomprometidos, mesmo na era da HAART. O presente estudo teve como objetivo avaliar a influência da carga viral do CMV e seu genótipo gB sobre a evolução de pacientes com AIDS. MÉTODOS: Amostras de sangue de 101 pacientes com AIDS foram coletadas e testadas para carga viral de HIV, a contagem de células CD4 e patógenos oportunistas, incluindo o CMV. Um sistema de PCRs seminested foi utilizado para detectar o genoma do CMV e em amostras positivas a carga viral de CMV e genotipagem foram estabelecidos por restrição enzimática e PCR em tempo real, respectivamente. Todos os pacientes foram acompanhados clinicamente durante quatro anos. RESULTADOS: Trinta pacientes (31 por cento) tiveram CMV detectado e todos os casos fatais (n = 5) ocorreram em pacientes deste grupo (p = 0,007), porém apenas dois pacientes tinham doença por CMV (1,9 por cento). No entanto, a carga viral não foi associada estatisticamente a nenhum dos parâmetros analisados. O genótipo de CMV mais freqüentemente observado foi gB2 (45,16 por cento), seguido por gB3 (35,48 por cento). O genótipo gB2 foi mais freqüente em pacientes com contagens abaixo de 200 células/mm³ CD4cell (p = 0,0017), e quase todos os casos fatais (80 por cento) tinham o genótipo gB2. CONCLUSÃO: Nosso estudo sugere que CMV e seu polimorfismo em genes relevantes biologicamentes, como a gB, pode ainda influenciar no prognóstico e evolução de pacientes com AIDS. O genótipo gB2 foi associado ao mau prognóstico do paciente.


Subject(s)
Adult , Female , Humans , Male , AIDS-Related Opportunistic Infections/virology , Cytomegalovirus Infections/virology , Cytomegalovirus/genetics , Leukocytes/virology , Viral Envelope Proteins/genetics , Genotype , Polymerase Chain Reaction , Prognosis , Viral Load
20.
Rev. chil. infectol ; 28(2): 113-117, abr. 2011. ilus
Article in Spanish | LILACS | ID: lil-592092

ABSTRACT

Background: Cytomegalovirus (CMV) infections are an important cause of morbidity and mortality in transplant recipient. To date, the antigenemia assay is the most used technique for diagnostic and management of CM V infections. However, quantification of CMV viral load by real time polymerase chain reaction (RT-PCR) has becoming the method of choice to detect CMV in a rapid, sensitive and specific manner. Objective: To compare antigenemia and RT-PCR assays in the detection of CMV in blood sample from solid organ and bone marrow transplant (BMT) in children attended at the Dr. Luis Calvo Mackenna Hospital. Methods: In a prospective study, we detect the presence of CMV in blood sample by RT-PCR and antigenemia assays. Results: We analyzed 219 blood samples from 68 children subjected to kidney, liver and BMT. Out of 219 samples analyzed, 147 were negative and 33 were positive for CMV by both techniques. Thirty-seven samples were positive only by RT-PCR and 2 by antigenemia. Considering the antigenemia as a reference, RT-PCR shows 94 percent, 80 percent, 34 percent and 99 percent sensitivity, specificity, positive and negative predictive values, respectively. The kappa coefficient between both techniques was 0.528. Conclusion: Quantitative determination of CMV viral load by RT-PCR is a sensitive technique with excellent negative predictive valué compared to antigenemia. Our results support the use of RT-PCR as a technique that might facilítate the diagnostic and treatment of active CMV infection in pediatric transplants.


Antecedentes: Las infecciones por citomegalovirus (CMV) corresponden a una importante causa de morbilidad y mortalidad en pacientes sometidos a trasplantes. Hasta la fecha, la detección de CMV en células infectadas en sangre periférica mediante la técnica de inmunofluorescencia (antigenemia) es la más utilizada para el diagnóstico y monitorización de la infección por este agente. Sin embargo, en el último tiempo la cuantificación de la carga de ácido nucleico (ADN) de CMV en sangre mediante la técnica de reacción de polimerasa en cadena en tiempo real (RPC-TR) ha permitido la detección de CMV de forma más rápida, sensible y específica. Objetivos: Comparar las técnicas de antigenemia y RPC-TR para la detección de CMV en sangre en niños sometidos a trasplante de órganos sólidos y trasplante de precursores hematopoyéticos (TPH) en el Hospital Dr. Luis Calvo Mackenna. Metodología: En un estudio prospectivo de seguimiento preventivo de reactivación se detectó la presencia de CMV en muestras de sangre utilizando las técnicas de RPC-TR y antigenemia. Resultados: Se analizaron 219 muestras de sangre, correspondiente a 68 niños sometidos a trasplante de hígado, riñon y TPH. De las muestras analizadas, 147 fueron negativas y 33 positivas para CMV utilizando ambas técnicas. Treinta y siete muestras resultaron ser positivas sólo por RPC-TR y dos sólo por antigenemia. Tomando en cuenta la antigenemia como referencia, la RPC-TR mostró una sensibilidad, especificidad, valor predictor positivo y negativo de 94 por ciento, 80 por ciento, 34 por ciento y 99 por ciento, respectivamente. El índice de concordancia entre ambas técnicas tuvo un valor de kappa = 0,528. Conclusión: La determinación cuantitativa de ADN de CMV por RPC-TR es una técnica sensible, con un gran valor predictor negativo comparada con la antigenemia. Los resultados obtenidos en este trabajo apoyan el uso de RPC-TR para el diagnóstico y tratamiento oportuno de las infecciones activas por CMV en niños sometidos a trasplantes.


Subject(s)
Child , Child, Preschool , Female , Humans , Male , Antigens, Viral/blood , Cytomegalovirus Infections/diagnosis , Cytomegalovirus/isolation & purification , Hematopoietic Stem Cell Transplantation , Organ Transplantation , Polymerase Chain Reaction/methods , Cytomegalovirus Infections/etiology , Cytomegalovirus Infections/immunology , Cytomegalovirus/genetics , Cytomegalovirus/immunology , DNA, Viral/blood , Postoperative Complications , Prospective Studies , Sensitivity and Specificity
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