RESUMO
First-void urine (FVU) samples, containing human papillomavirus (HPV)-specific IgG from female genital tract secretions, provide a non-invasive option for disease monitoring and vaccine impact assessment. This study explores the utility of FVU for IgG quantification, exploring stability and compatibility with DNA preservation methods, alongside various IgG enrichment methods. Healthy female volunteers provided FVU and serum samples. FVU was collected with or without urine conservation medium (UCM) and stored under different conditions before freezing at -80 °C. Four IgG enrichment methods were tested on FVU samples. All samples were analyzed using three total human IgG quantification assays and an in-house HPV16-specific IgG assay. Samples stored with UCM buffer had higher total and HPV16-specific IgG concentrations (p ≤ 0.01) and IgG remained stable for at least 14 days at room temperature. Among IgG enrichment methods, Amicon filtration (AM) and AM combined with Melon Gel purification (AM-MG) provided similar HPV16-IgG concentrations, correlating strongly with serum levels. Protein G magnetic beads methods were incompatible with time-resolved fluorescence-based assays. This study highlights FVU as a reliable and convenient sample for IgG quantification, demonstrating stability for at least 14 days at room temperature and compatibility with UCM DNA preservation. It emphasizes the need to select appropriate IgG enrichment methods and confirms the suitability of both AM and AM-MG methods, with a slightly better performance for AM-MG.
Assuntos
Papillomavirus Humano 16 , Imunoglobulina G , Humanos , Feminino , Papillomavirus Humano 16/imunologia , Imunoglobulina G/sangue , Imunoglobulina G/urina , Adulto , Anticorpos Antivirais/sangue , Anticorpos Antivirais/urina , Anticorpos Antivirais/imunologia , Infecções por Papillomavirus/urina , Infecções por Papillomavirus/virologia , Infecções por Papillomavirus/diagnóstico , Infecções por Papillomavirus/imunologia , Adulto JovemRESUMO
The potential of first-void (FV) urine as a non-invasive method to monitor human papillomavirus (HPV) vaccination has been reported, mainly focusing on urine as a sample to assess HPV DNA. Besides HPV DNA, vaccine-induced HPV antibodies originating from cervicovaginal secretions were recently shown to be detectable in FV urine as well. This presents a novel opportunity for non-invasive sampling to monitor HPV antibody status in women participating in large epidemiological studies and HPV vaccine trials. The simultaneous assessment of both HPV infection and immunogenicity on a non-invasive, readily obtained sample is particularly attractive.
Assuntos
Alphapapillomavirus/imunologia , Anticorpos Antivirais/urina , Imunogenicidade da Vacina , Infecções por Papillomavirus/diagnóstico , Vacinas contra Papillomavirus/uso terapêutico , Biomarcadores/urina , Feminino , Humanos , Infecções por Papillomavirus/imunologia , Infecções por Papillomavirus/urina , Infecções por Papillomavirus/virologia , Valor Preditivo dos Testes , Reprodutibilidade dos Testes , UrináliseRESUMO
IntroductionCrimean-Congo haemorrhagic fever (CCHF) is a tick-borne disease in Africa, Asia, the Balkan peninsula, the south-east of Europe and the Middle East, with mortality rates of 3-30%. Transmission can also occur through contact with infected animals or humans.AimThis observational, prospective case series aimed to investigate detectable viral genomic RNA in whole-body fluids and antibody dynamics in consecutive daily samples of patients diagnosed with CCHF until discharge from hospital.MethodsWe tested 18 patients and 824 swabs and sera with RT-PCR and 125 serum samples serologically.ResultsThe longest duration until clearance of viral RNA was 18 days from serum collection and 18, 15, 13, 19 and 17 days, respectively, from nasal, oral, genital (urethral or vaginal) and faecal swab, and urine. In seven patients, viral load decreased in serum at the same time as it increased in urine or persisted at the same logarithmic values. Despite clearance in serum, viral RNA was detected in faeces and genital swabs in two and three patients, respectively. Viral clearance from body fluids occurred earlier than from serum in eight patients on ribavirin treatment. The shortest seroconversion time was 3 days after symptom onset for IgM and IgG. Seroconversion of IgG occurred until Day 14 of symptoms.ConclusionWe report persistence of viral RNA in urine, faeces and genital swabs despite serum clearance. This may indicate a need for extending isolation precautions, re-evaluating discharge criteria and transmission risk after discharge, and considering oral swabs as a less invasive diagnostic alternative.
Assuntos
Vírus da Febre Hemorrágica da Crimeia-Congo/isolamento & purificação , Febre Hemorrágica da Crimeia/diagnóstico , Eliminação de Partículas Virais , Adolescente , Adulto , Idoso , Anticorpos Antivirais/sangue , Anticorpos Antivirais/urina , Antivirais/uso terapêutico , Criança , Feminino , Genoma Viral , Vírus da Febre Hemorrágica da Crimeia-Congo/genética , Febre Hemorrágica da Crimeia/tratamento farmacológico , Febre Hemorrágica da Crimeia/epidemiologia , Humanos , Imunoglobulina G/sangue , Imunoglobulina M/sangue , Masculino , Pessoa de Meia-Idade , Estudos Prospectivos , RNA Viral/sangue , RNA Viral/urina , Ribavirina/uso terapêutico , Testes Sorológicos , Doenças Transmitidas por Carrapatos , Turquia/epidemiologia , Carga Viral , Adulto JovemRESUMO
BACKGROUND: Dengue is an important arboviral disease with about 100 million dengue cases per year, of which, ~5% result in severe disease. Clinical differentiation of dengue from other acute febrile illnesses (AFI) is difficult, and diagnostic blood tests are costly. We evaluated the utility of anti-DENV IgM in urine to identify dengue cases among AFI patients enrolled in a clinical study. METHODS: Between May 2012-March 2013, 1538 study participants with fever for ≤7 days were enrolled, a medical history was obtained, and serum and urine specimens were collected. Serum was tested for DENV RNA and anti-DENV IgM. Urine was tested for anti-DENV IgM, and its sensitivity and specificity to detect sera laboratory-positive dengue cases were calculated. We evaluated if urine anti-DENV IgM positivity early (≤5 days post-illness onset [DPO]) and late (6-14 DPO) in the clinical course was associated with dengue severity. RESULTS: Urine anti-DENV IgM sensitivity and specificity were 47.4% and 98.5%, respectively, when compared with serum anti-DENV IgM ELISA results, and 29.7% and 91.1% when compared with serum rRT-PCR results. There was no correlation between urine anti-DENV IgM positivity and patient sex or pre-existing chronic disease. Early in the clinical course, a significantly higher proportion of those who developed dengue with warning signs had anti-DENV IgM in their urine when compared to those without warning signs (20.4% vs. 4.3%). There was no difference in the proportion with urine anti-DENV IgM positivity between severity groups late in the clinical course. CONCLUSION: While detection of urine anti-DENV IgM lacked adequate diagnostic sensitivity, it is a highly specific marker for laboratory-positive dengue, and its presence early in the clinical course may distinguish those with more severe disease. Further assessment of urine anti-DENV IgM by DPO is warranted to determine its utility as an early diagnostic (and possibly prognostic) marker for dengue.
Assuntos
Anticorpos Antivirais/urina , Vírus da Dengue/imunologia , Dengue/diagnóstico , Dengue/urina , Imunoglobulina M/urina , Adolescente , Adulto , Criança , Dengue/epidemiologia , Testes Diagnósticos de Rotina/métodos , Feminino , Febre/diagnóstico , Humanos , Masculino , Pessoa de Meia-Idade , Porto Rico/epidemiologia , Sensibilidade e Especificidade , Adulto JovemRESUMO
Rapid point-of-care testing is a megatrend in infectious disease diagnosis. We have introduced a homogeneous immunoassay concept, which is based on the simultaneous binding of antigen and protein L to a given immunoglobulin molecule. The complex formation is detected utilizing time-resolved Förster resonance energy transfer between antigen-attached donor and acceptor-labeled protein L, hence the name LFRET. Here, we demonstrate that urine can be used as a sample matrix in LFRET-based serodiagnostics. We studied urine samples collected during the hospitalization and recovery of patients with acute Puumala orthohantavirus (PUUV) infection. We compared PUUV antibody-specific LFRET signals in urine to those in plasma, and found excellent correlation in the test outcomes The LFRET test from urine was positive in 40/40 patients with acute PUUV infection. PUUV causes a mild form of hemorrhagic fever with renal syndrome, characterized by acute kidney injury and proteinuria. Immunofluorescence and western blotting demonstrated PUUV-IgG and -IgA in urine, however, the presence of intact immunoglobulins did not fully explain the LFRET signals. We purified free light chains (FLCs) from both urine and serum of healthy volunteers and patients with acute PUUV infection, and verified the presence of antigen-specific FLCs. Antigen-specific FLCs provide a new means for non-invasive antibody detection and disease diagnosis.
Assuntos
Infecções por Hantavirus/diagnóstico , Cadeias Leves de Imunoglobulina/urina , Orthohantavírus/isolamento & purificação , Testes Sorológicos/métodos , Anticorpos Antivirais/urina , Proteínas do Capsídeo/imunologia , Orthohantavírus/imunologia , Infecções por Hantavirus/urina , Febre Hemorrágica com Síndrome Renal/diagnóstico , Febre Hemorrágica com Síndrome Renal/urina , Humanos , Imunoensaio , Imunoglobulina A/urina , Imunoglobulina G/urina , Cadeias Leves de Imunoglobulina/imunologia , Testes Imediatos , Virus Puumala/imunologia , Virus Puumala/isolamento & purificação , Proteínas do Core Viral/imunologiaAssuntos
Camelus/urina , Infecções por Coronavirus/veterinária , Coronavírus da Síndrome Respiratória do Oriente Médio/isolamento & purificação , RNA Viral/urina , Animais , Anticorpos Antivirais/urina , Infecções por Coronavirus/urina , Infecções por Coronavirus/virologia , Reservatórios de Doenças/veterinária , Reservatórios de Doenças/virologia , Coronavírus da Síndrome Respiratória do Oriente Médio/genética , ZoonosesRESUMO
BACKGROUND: Monitoring HPV antibodies non-invasively would be a major advantage for large epidemiological studies and follow-up of vaccinees. OBJECTIVES: This study investigated the presence of HPV-specific antibody transudates from systemic circulation in first-void urine of (un)vaccinated subjects and the agreement with paired sera. STUDY DESIGN: In this case-control study, 55 paired first-void urine and serum samples were included from 19- to 26-year-old women, unvaccinated (n = 19) or vaccinated (n = 36) with the bi- or quadrivalent HPV vaccine during adolescence (NCT02714114). Human IgA, total human IgG, and HPV6/11/16/18-Ig(M/G/A) were measured in paired samples. RESULTS: Significant positive Spearman rank correlations (rs) were found in HPV-specific antibody levels between paired samples (HPV6: rs = 0.777; HPV11: rs = 0.757; HPV16: rs = 0.876; HPV18: rs = 0.636 (p < 0.001)). In both first-void urine and serum, significantly higher HPV6/11/16/18 antibody levels were observed in vaccinated compared with unvaccinated women (p ≤ 0.017). CONCLUSIONS: The present study provides the first proof that vaccine-induced HPV antibodies are detectable in the first-void urine of young women. Moreover, significant positive correlations were observed between HPV6/11/16/18-antibodies in first-void urine and paired sera. Further optimization and validation are required to demonstrate its potential use in epidemiological studies and follow-up of HPV vaccination.
Assuntos
Anticorpos Antivirais/urina , Secreções Corporais/virologia , Papillomaviridae/imunologia , Infecções por Papillomavirus/prevenção & controle , Vacinas contra Papillomavirus/imunologia , Anticorpos Antivirais/sangue , Estudos de Casos e Controles , Colo do Útero/virologia , Feminino , Papillomavirus Humano 11/imunologia , Papillomavirus Humano 16/imunologia , Papillomavirus Humano 18/imunologia , Humanos , Imunoglobulina A/sangue , Imunoglobulina A/urina , Imunoglobulina G/sangue , Imunoglobulina G/urina , Biópsia Líquida , Infecções por Papillomavirus/imunologia , Infecções por Papillomavirus/urina , Vacinação , Vagina/virologia , Adulto JovemRESUMO
BACKGROUND: A range of different neurological manifestations has been reported in fetuses and adults after Zika virus (ZIKV) infection. OBJECTIVE: We describe a detection of the ZIKV in the brain tissue from a multiple sclerosis (MS) patient with acute disseminated encephalomyelitis (ADEM)-like event in Rio de Janeiro, Brazil. METHODS: Biological samples collected during the hospitalization were tested by serology and molecular diagnostic for various infectious agents. Histopathological analysis was performed using the anti-flavivirus group 4G2 monoclonal antibody, anti-ZIKV non-structural 1 (NS1) monoclonal antibody, and anti-CD4, CD8, and CD11b antibodies. RESULTS: Anti-ZIKV IgM and IgG antibodies were positive in the serum and urine. A brain biopsy showed ZIKV protein in brain cells and T CD8 infiltration in brain tissue. CONCLUSION: Our data describe the coexistence of a recent central nervous system (CNS) ZIKV infection accompanied by a severe ADEM-like syndrome outcome in a patient with clinical history of MS. A de novo immune response concomitant with ZIKV infection might be involved in the mechanism of the ADEM-like syndrome and response to immunotherapy. The present report reinforces the importance of providing the differential diagnosis of acute episodes of MS exacerbation in an environment prone to ZIKV expression.
Assuntos
Encéfalo/microbiologia , Encefalomielite Aguda Disseminada/diagnóstico , Esclerose Múltipla Recidivante-Remitente , Infecção por Zika virus/diagnóstico , Zika virus/isolamento & purificação , Adulto , Anticorpos Antivirais/sangue , Anticorpos Antivirais/urina , Encefalomielite Aguda Disseminada/microbiologia , Feminino , Humanos , Infecção por Zika virus/sangue , Infecção por Zika virus/imunologia , Infecção por Zika virus/urinaRESUMO
A 27-year-old pregnant woman who was positive for anti-cytomegalovirus (CMV) antibodies gave birth to a congenitally CMV-infected neonate at 40 weeks of gestation. According to strain-specific serological analysis, which is able to determine the two types of CMV glycoprotein H (gH), the mother possessed anti-gH(To) antibodies only, but the neonate possessed anti-gH(AD) and anti-gH(To) antibodies at 4 weeks after birth. As the anti-gH(To) IgG was decreased in the neonate at 8 months post-delivery, these antibodies are thought to have been transferred from the mother as maternal antibodies. The anti-gH(AD) IgG level was maintained in the child even after 8 months post-delivery. Congenital infection with a CMV gH(AD) type strain was confirmed by strain-specific real-time PCR using a urine specimen from the child. On the other hand, anti-gH(AD) IgG was not detected even after 8 months post-delivery in a maternal specimen. The mother only produced antibodies against the CMV strain identified as the primary infection, which is characteristic of original antigenic sin.
Assuntos
Anticorpos Antivirais/urina , Infecções por Citomegalovirus/diagnóstico , Transmissão Vertical de Doenças Infecciosas , Complicações Infecciosas na Gravidez/diagnóstico , Adulto , Citomegalovirus/isolamento & purificação , Citomegalovirus/metabolismo , Infecções por Citomegalovirus/urina , Feminino , Humanos , Imunoglobulina G/urina , Lactente , Gravidez , Complicações Infecciosas na Gravidez/urina , Reação em Cadeia da Polimerase em Tempo Real , Manejo de Espécimes , Proteínas do Envelope Viral/urinaRESUMO
OBJECTIVE: We evaluated cytomegalovirus (CMV) immunoglobulin M (IgM) titer in pregnant women with primary infection as a predictive factor for congenital infection. STUDY DESIGN: Maternal CMV antibody screening during the first trimester was conducted prospectively at 16 centers in Japan between September 2013 and 2015. Women with confirmed maternal primary infection underwent testing for fetal congenital infection, and we investigated the positive predictive value of CMV IgM titer levels for congenital infection in women with a low IgG avidity. RESULTS: We identified 6 (8.6%) cases of congenital infection among 70 pregnant women with positive/borderline IgG, positive IgM and IgG avidity index ⩽35.0% and 11 (39.3%) among 28 women with IgG and/or IgM seroconversion. IgM titer level ⩾6.00 index showed the highest positive predictive value (17.1%). CONCLUSION: High titer of CMV IgM during the first trimester in pregnant women with primary infection is a risk factor for congenital infection.
Assuntos
Anticorpos Antivirais/urina , Infecções por Citomegalovirus/sangue , Citomegalovirus/imunologia , Imunoglobulina M/sangue , Complicações Infecciosas na Gravidez/sangue , Primeiro Trimestre da Gravidez/sangue , Adolescente , Adulto , Estudos de Casos e Controles , Infecções por Citomegalovirus/congênito , Infecções por Citomegalovirus/diagnóstico , Ensaio de Imunoadsorção Enzimática , Feminino , Humanos , Imunoglobulina G/sangue , Imunoglobulina G/imunologia , Imunoglobulina M/imunologia , Recém-Nascido , Pessoa de Meia-Idade , Valor Preditivo dos Testes , Gravidez , Complicações Infecciosas na Gravidez/imunologia , Primeiro Trimestre da Gravidez/imunologia , Estudos Prospectivos , Reação em Cadeia da Polimerase em Tempo Real , Adulto JovemRESUMO
Natalizumab greatly reduces inflammatory relapses in multiple sclerosis (MS) by blocking the integrin-mediated leukocyte traffic to the brain, but less is known about its effects on the systemic immunity. We measured 48 cytokines/chemokines in sera from 19 natalizumab-treated MS patients. Serum concentrations of both anti-(IL-10, IL1ra) and pro-inflammatory (IL7, IL16) molecules decreased after 21-month treatment, without associations to unbalanced Th2/Th1cytokine ratios, clinical responses, and blood/urine replication of polyomavirus JC (JCPyV). No patient developed the JCPyV-related progressive multifocal leukoencephalopathy (PML), the major risk factor of natalizumab therapy. Our data suggest that natalizumab has marginal impact on the systemic immunity.
Assuntos
Citocinas/sangue , Fatores Imunológicos/uso terapêutico , Esclerose Múltipla/sangue , Esclerose Múltipla/tratamento farmacológico , Natalizumab/uso terapêutico , Adulto , Anticorpos Antivirais/sangue , Anticorpos Antivirais/urina , Feminino , Seguimentos , Humanos , Peptídeos e Proteínas de Sinalização Intercelular/metabolismo , Vírus JC/genética , Vírus JC/imunologia , Masculino , Subpopulações de Linfócitos T/efeitos dos fármacos , Subpopulações de Linfócitos T/imunologia , Fatores de TempoRESUMO
The kinetics of dengue virus (DENV)-specific IgA antibody in urine and the potential correlation with disease severity remain elusive. In this study, 262 serial urine samples from 78 laboratory-confirmed patients were assayed by a commercial immunoglobulin A (IgA) kit against DENV. All cases were classified into dengue fever (DF) and severe dengue (SD) according to the 2009 WHO/TDR guideline. The total positive rate of IgA in urine was 59%. DENV-specific IgA was detected in urine from day 2 to day 13 after the onset of illness in DF patients; While for SD patients, anti-DENV IgA could be detected till day 14. The positive rate of IgA in patients with secondary infection was higher than that in patients with primary infection. Importantly, during 4-7 days after the onset of illness, the IgA positive rate of SD patients was significantly higher than that of DF patients. Especially, the intensity of IgA signal in SD patients was obviously stronger than that in DF patient at the recovery stage. Overall, our results suggested that the existence of DENV-specific IgA antibodies in urine might be a warning sign for the severity of disease and its measurement might provide valuable guidance for proper patient management.
Assuntos
Anticorpos Antivirais/urina , Vírus da Dengue/imunologia , Dengue/imunologia , Dengue/patologia , Imunoglobulina A/urina , Adulto , Idoso , Idoso de 80 Anos ou mais , China , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Adulto JovemRESUMO
Hendra virus (HeV) is an important emergent virus in Australia known to infect horses and humans in certain regions of the east coast. Whilst pteropid bats ("flying foxes") are considered the natural reservoir of HeV, which of the four mainland species is the principal reservoir has been a source of ongoing debate, particularly as shared roosting is common. To help resolve this, we sampled a colony consisting of just one of these species, the grey-headed flying fox, (Pteropus poliocephalus), at the southernmost extent of its range. Using the pooled urine sampling technique at approximately weekly intervals over a two year period, we determined the prevalence of HeV and related paramyxoviruses using a novel multiplex (Luminex) platform. Whilst all the pooled urine samples were negative for HeV nucleic acid, we successfully identified four other paramyxoviruses, including Cedar virus; a henipavirus closely related to HeV. Collection of serum from individually caught bats from the colony showed that antibodies to HeV, as estimated by a serological Luminex assay, were present in between 14.6% and 44.5% of animals. The wide range of the estimate reflects uncertainties in interpreting intermediate results. Interpreting the study in the context of HeV studies from states to the north, we add support for an arising consensus that it is the black flying fox and not the grey-headed flying fox that is the principal source of HeV in spillover events to horses.
Assuntos
Quirópteros/virologia , Vírus Hendra/fisiologia , Infecções por Henipavirus/virologia , Doenças dos Cavalos/virologia , Cavalos/virologia , Animais , Anticorpos Antivirais/sangue , Anticorpos Antivirais/imunologia , Anticorpos Antivirais/urina , Austrália/epidemiologia , Reservatórios de Doenças/virologia , Geografia , Vírus Hendra/imunologia , Infecções por Henipavirus/epidemiologia , Infecções por Henipavirus/transmissão , Interações Hospedeiro-Patógeno , Humanos , Infecções por Paramyxoviridae/epidemiologia , Infecções por Paramyxoviridae/transmissão , Infecções por Paramyxoviridae/virologia , Paramyxovirinae/imunologia , Paramyxovirinae/fisiologia , Prevalência , Estações do Ano , Fatores de Tempo , Zoonoses/virologiaRESUMO
Sixty-three natalizumab-treated patients with relapsing multiple sclerosis were screened for JC polyomavirus (JCV) viruria. Urinary-positive patients were longitudinally sampled for up to 24 weeks. Using methods that distinguish encapsidated virus from naked viral DNA, 17.5 % of patients were found to excrete virus, consistent with the prevalence of urinary excretion in the general population. Unexpectedly, urinary excretion was predominantly seen (>73 %) in patients with high JC antibody index (≥2.0). Active JCV infection, therefore, tends to occur in natalizumab patients that carry a high risk factor for the development of disease, directly linking JC infection to the risk factors for PML development.
Assuntos
DNA Viral/urina , Fatores Imunológicos/uso terapêutico , Vírus JC/patogenicidade , Leucoencefalopatia Multifocal Progressiva/tratamento farmacológico , Esclerose Múltipla Recidivante-Remitente/tratamento farmacológico , Natalizumab/uso terapêutico , Anticorpos Antivirais/urina , Humanos , Vírus JC/imunologia , Leucoencefalopatia Multifocal Progressiva/etiologia , Leucoencefalopatia Multifocal Progressiva/urina , Leucoencefalopatia Multifocal Progressiva/virologia , Estudos Longitudinais , Esclerose Múltipla Recidivante-Remitente/complicações , Esclerose Múltipla Recidivante-Remitente/urina , Esclerose Múltipla Recidivante-Remitente/virologia , Fatores de Risco , UrináliseRESUMO
BACKGROUND: Dengue laboratory diagnosis is essentially based on detection of the virus, its components or antibodies directed against the virus in blood samples. Blood, however, may be difficult to draw in some patients, especially in children, and sampling during outbreak investigations or epidemiological studies may face logistical challenges or limited compliance to invasive procedures from subjects. The aim of this study was to assess the possibility of using saliva and urine samples instead of blood for dengue diagnosis. METHODOLOGY/PRINCIPAL FINDINGS: Serial plasma, urine and saliva samples were collected at several time-points between the day of admission to hospital until three months after the onset of fever in children with confirmed dengue disease. Quantitative RT-PCR, NS1 antigen capture and ELISA serology for anti-DENV antibody (IgG, IgM and IgA) detection were performed in parallel on the three body fluids. RT-PCR and NS1 tests demonstrated an overall sensitivity of 85.4%/63.4%, 41.6%/14.5% and 39%/28.3%, in plasma, urine and saliva specimens, respectively. When urine and saliva samples were collected at the same time-points and tested concurrently, the diagnostic sensitivity of RNA and NS1 detection assays was 69.1% and 34.4%, respectively. IgG/IgA detection assays had an overall sensitivity of 54.4%/37.4%, 38.5%/26.8% and 52.9%/28.6% in plasma, urine and saliva specimens, respectively. IgM were detected in 38.1% and 36% of the plasma and saliva samples but never in urine. CONCLUSIONS: Although the performances of the different diagnostic methods were not as good in saliva and urine as in plasma specimens, the results obtained by qRT-PCR and by anti-DENV antibody ELISA could well justify the use of these two body fluids to detect dengue infection in situations when the collection of blood specimens is not possible.
Assuntos
Anticorpos Antivirais/análise , Vírus da Dengue/isolamento & purificação , Dengue/diagnóstico , Dengue/epidemiologia , Epidemias , Proteínas não Estruturais Virais/análise , Adolescente , Anticorpos Antivirais/sangue , Anticorpos Antivirais/urina , Camboja , Criança , Pré-Escolar , Dengue/sangue , Dengue/urina , Vírus da Dengue/genética , Vírus da Dengue/imunologia , Testes Diagnósticos de Rotina , Progressão da Doença , Ensaio de Imunoadsorção Enzimática , Feminino , Genoma Viral , Humanos , Isotipos de Imunoglobulinas/análise , Isotipos de Imunoglobulinas/sangue , Isotipos de Imunoglobulinas/urina , Masculino , Plasma/química , Plasma/imunologia , Plasma/virologia , Reação em Cadeia da Polimerase , RNA Viral/isolamento & purificação , Saliva/química , Saliva/imunologia , Saliva/virologia , Urina/química , Urina/fisiologia , Urina/virologia , Proteínas não Estruturais Virais/sangue , Proteínas não Estruturais Virais/urinaRESUMO
BACKGROUND: Urine is an important source for the detection of infections caused by CMV in stem cell transplant patients. Currently, there is no agreement about the type of urine specimen. In order to investigate which is the better specimen type for quantitative detection of CMV, we compared the results from urine supernatant and sediment from the same patients. METHODS: Seventy urine specimens were collected from patients with hematological disorders or solid tumors. After performing shell vial culture, residual urine specimens were centrifuged. Then, 10 mL of each urine supernatant and sediment were taken and immediately frozen at -70 degrees C. Afterwards, archived urine specimens were thawed at room temperature and CMV-quantitative PCR was performed on both the supernatant and sediment fraction of urine. The results from each patient were reviewed for CMV antigenemia, blood shell vial culture, CMV-IgM or IgG, and clinical symptoms. RESULTS: CMV-qPCR results for the urine sediment fraction revealed a significant difference (p = 0.012) between the active CMV infection group and the latent CMV infection group. In addition, receiver operating characteristic curves for active CMV infection revealed that CMV-qPCR using urine sediment produced more accurate results than urine supernatant. CONCLUSIONS: These findings suggest that the sediment fraction of urine is a more suitable specimen in CMV-qPCR testing.
Assuntos
Infecções por Citomegalovirus/urina , Citomegalovirus/isolamento & purificação , Transplante de Células-Tronco/efeitos adversos , Urinálise/métodos , Viremia/urina , Adolescente , Adulto , Anticorpos Antivirais/urina , Antígenos Virais/urina , Criança , Pré-Escolar , Citomegalovirus/genética , Feminino , Neoplasias Hematológicas/terapia , Neoplasias Hematológicas/urina , Humanos , Imunoglobulina G/urina , Imunoglobulina M/urina , Lactente , Masculino , Pessoa de Meia-Idade , Neoplasias/terapia , Neoplasias/urina , Reação em Cadeia da Polimerase em Tempo Real , Manejo de Espécimes , Adulto JovemRESUMO
The risk of developing progressive multifocal leukoencephalopathy (PML), as a consequence of infection/reactivation with JC virus (JCV), is consistent in natalizumab-treated multiple sclerosis (MS) patients, with 430 cases of PML reported so far. The risk of PML is higher in JCV seropositive patients, and it is recommended that only MS patients without JCV antibodies should be enrolled in the treatment postulating that they do not have JCV infection.We have studied forty-two natalizumab-treated MS patients, and urine and blood were collected monthly for up to 60 months. JCV and BK virus (BKV) DNA presence was verified using quantitative real-time PCR assays, and serum anti-JCV antibodies were measured with the Stratify and/or Stratify DxSelect tests.JCV and BKV DNA were not found in the blood samples, whereas they were found at least once in the urine of 21 of 42 (50 %) and of 25/42 (59.5 %) patients, respectively. JCV DNA urinary shedding increased up to month 24 of natalizumab treatment (45.2 %), and the effect of time was significant for JCV (p = 0.04), but not for BKV (p = 0.39). JCV viruria and seropositivity did not completely correlate, since three patients shedding JCV DNA in the urine were seronegative according to the serological tests.The results indicated that natalizumab therapy may increase the rate of JCV urinary shedding. Additionally, we confirmed that the identification of JCV carriers cannot solely rely on serological tests, but sensitive methods for viral DNA detection should be adopted to more precisely identify the truly JCV uninfected cases.
Assuntos
Anticorpos Antivirais/urina , DNA Viral/urina , Fatores Imunológicos/uso terapêutico , Esclerose Múltipla Recidivante-Remitente/tratamento farmacológico , Esclerose Múltipla Recidivante-Remitente/virologia , Natalizumab/uso terapêutico , Adulto , Anticorpos Antivirais/sangue , DNA Viral/sangue , Ensaio de Imunoadsorção Enzimática , Feminino , Humanos , Vírus JC/isolamento & purificação , Masculino , Pessoa de Meia-Idade , Infecções por Polyomavirus/virologia , Reação em Cadeia da Polimerase em Tempo Real , Estudos Soroepidemiológicos , Eliminação de Partículas Virais/fisiologia , Adulto JovemRESUMO
OBJECTIVE: To explore the cost-effectiveness of the diagnosis of congenital cytomegalovirus (CMV) infection by fluorescent quantitative polymerase chain reaction (FQ-PCR) in neonates. METHODS: Serum CMV immunoglobulin M (CMV-IgM) and CMV-IgG were detected using ELISA in 610 neonates aged less than 14 days. CMV DNA content was detected by FQ-PCR. The cost-effectiveness analysis was then performed. RESULTS: The positive rate of FQ-PCR in neonates with positive CMV-IgM was 42.9% (15/35), while, 2.9% (16/547) in neonates with positive CMV-IgG. The mean logarithm values of CMV DNA in neonates with positive CMV-IgM were higher than those in neonates with positive CMV-IgG (5.79±1.24 vs 4.11±0.87; P<0.01). The costs of the diagnosis of CMV infection by FQ-PCR were 256 RMB/case in neonates with positive CMV-IgM, and 3 760 RMB/case in neonates with positive CMV-IgG. CONCLUSIONS: The CMV DNA content in neonates with positive CMV-IgM is higher than that in neonates with positive CMV-IgG. Diagnosis of congenital CMV infection by FQ-PCR in neonates with positive CMV-IgG is not suitable for large scale epidemiological survey because of high cost-effectiveness ratio.
Assuntos
Infecções por Citomegalovirus/congênito , Infecções por Citomegalovirus/diagnóstico , Reação em Cadeia da Polimerase/economia , Anticorpos Antivirais/urina , Ensaio de Imunoadsorção Enzimática , Feminino , Fluorescência , Humanos , Imunoglobulina G/urina , Imunoglobulina M/urina , Recém-Nascido , Masculino , Reação em Cadeia da Polimerase/métodosRESUMO
OBJECTIVE: Analyses were conducted to determine the clinical utility of measuring JC virus (JCV) DNA in blood or urine of natalizumab-treated multiple sclerosis (MS) patients to predict the risk of progressive multifocal leukoencephalopathy (PML). METHODS: A total of 12,850 blood and urine samples from nearly 1,400 patients participating in natalizumab clinical trials were tested for JCV DNA using a commercially available quantitative polymerase chain reaction (qPCR) assay. A subset of these samples was also tested using a more sensitive qPCR assay developed at the National Institutes of Health (NIH). RESULTS: At the time natalizumab dosing was suspended, JCV DNA was detected in plasma by the commercial assay in 4 of 1,397 (0.3%) patients; the NIH assay confirmed these positive samples and detected JCV DNA in an additional 2 of 205 (1%) patients who tested negative with the commercial assay. None of these 6 JCV DNA positive patients developed PML. In a 48-week study testing the safety of natalizumab redosing, JCV DNA was detected in plasma of 6 of 1,094 (0.3%) patients, none of whom developed PML. Urine at baseline and week 48 was assessed in 224 patients; 58 (26%) were positive at baseline, and 55 (25%) were positive after 48 weeks of natalizumab, treatment. JCV DNA was not detected in peripheral blood mononuclear cells from any of these 1,094 patients before or after natalizumab treatment. In 5 patients who developed PML, JCV DNA was not detected in blood at any time point before symptoms first occurred. INTERPRETATION: Measuring JCV DNA in blood or urine with currently available methods is unlikely to be useful for predicting PML risk in natalizumab-treated MS patients.