RESUMO
An accurate urine test for diverse populations with active tuberculosis could be transformative for preventing TB deaths. Urinary liporabinomannan (LAM) testing has been previously restricted to HIV co-infected TB patients. In this study we evaluate urinary LAM in HIV negative, pediatric and adult, pulmonary and extrapulmonary tuberculosis patients. We measured 430 microbiologically confirmed pretreatment tuberculosis patients and controls from Peru, Guinea Bissau, Venezuela, Uganda and the United States using three monoclonal antibodies, MoAb1, CS35, and A194, which recognize distinct LAM epitopes, a one-sided immunoassay, and blinded cohorts. We evaluated sources of assay variability and comorbidities (HIV and diabetes). All antibodies successfully discriminated TB positive from TB negative patients. ROAUC from the average of three antibodies' responses was 0.90; 95% CI 0.87-0.93, 90% sensitivity, 73.5% specificity (80 pg/mL). MoAb1, recognizing the 5-methylthio-D-xylofuranose(MTX)-mannose(Man) cap epitope, performed the best, was less influenced by glycosuria and identified culture positive pediatric (N = 19) and extrapulmonary (N = 24) patients with high accuracy (ROAUC 0.87, 95% CI 0.77-0.98, 0.90 sensitivity 0.80 specificity at 80 pg/mL; ROAUC = 0.96, 95% CI 0.92-0.99, 96% sensitivity, 80% specificity at 82 pg/mL, respectively). The MoAb1 antibody, recognizing the MTX-Man cap epitope, is a novel analyte for active TB detection in pediatric and extrapulmonary disease.
Assuntos
Lipopolissacarídeos/análise , Tuberculose/diagnóstico , Tuberculose/imunologia , Adulto , Coinfecção/urina , Epitopos/imunologia , Feminino , Guiné-Bissau , Infecções por HIV/urina , Humanos , Imunoensaio/métodos , Testes Imunológicos/métodos , Lipopolissacarídeos/imunologia , Lipopolissacarídeos/urina , Masculino , Pessoa de Meia-Idade , Mycobacterium tuberculosis/imunologia , Peru , Sistemas Automatizados de Assistência Junto ao Leito , Sensibilidade e Especificidade , Tuberculose/classificação , Tuberculose Pulmonar/microbiologia , Uganda , Estados Unidos , VenezuelaRESUMO
Raltegravir (RAL) is a HIV-integrase inhibitor recommended for treatment of HIV type 1 infection during pregnancy. The elimination of RAL to RAL glucuronide (RAL GLU) is mediated primarily by UDP glucuronosyltransferase 1A1 (UGT1A1). The present study shows the development and validation of 4 different methods for the analysis of RAL and RAL GLU in plasma and in urine samples. The methods were applied to evaluate the maternal-fetal pharmacokinetics of RAL and RAL GLU in a HIV-infected pregnant woman receiving RAL 400â¯mg twice daily. The sample preparation for RAL and RAL GLU analysis in 25⯵L plasma and 100⯵L diluted urine (10-fold with water containing 0.1% formic acid) were carried out by protein precipitation procedure. RAL and RAL GLU generate similar product mass fragments and require separation in the chromatographic system, so a suitable resolution was achieved for unchanged RAL and RAL GLU employing Ascentis Express C18 (75â¯×â¯4.6â¯mm, 2.7⯵m) for both plasma and urine samples. The methods showed linearities at the ranges of 0.1-13.5⯵g/mL RAL and 0.15-19.5⯵g/mL RAL GLU in urine and 10-2000â¯ng/mL RAL and 2.5-800 RAL GLU in plasma. Precise and accurate evaluation showed coefficients of variation and relative errors ≤ 15%. The methods have been successfully applied in a maternal-fetal pharmacokinetic study.
Assuntos
Infecções por HIV/tratamento farmacológico , Inibidores de Integrase de HIV/análise , Troca Materno-Fetal , Complicações Infecciosas na Gravidez/tratamento farmacológico , Raltegravir Potássico/análise , Brasil , Cromatografia Líquida de Alta Pressão/métodos , Feminino , Glucuronídeos/administração & dosagem , Glucuronídeos/sangue , Glucuronídeos/química , Infecções por HIV/sangue , Infecções por HIV/urina , Inibidores de Integrase de HIV/administração & dosagem , Inibidores de Integrase de HIV/química , Inibidores de Integrase de HIV/farmacocinética , Humanos , Recém-Nascido , Permeabilidade , Placenta/metabolismo , Gravidez , Complicações Infecciosas na Gravidez/sangue , Complicações Infecciosas na Gravidez/urina , Terceiro Trimestre da Gravidez/metabolismo , Raltegravir Potássico/administração & dosagem , Raltegravir Potássico/química , Raltegravir Potássico/farmacocinética , Espectrometria de Massas em Tandem/métodos , Cordão Umbilical/químicaRESUMO
BACKGROUND: Early diagnosis of reactivated Chagas disease in HIV patients could be lifesaving. In Latin America, the diagnosis is made by microscopical detection of the T. cruzi parasite in the blood; a diagnostic test that lacks sensitivity. This study evaluates if levels of T. cruzi antigens in urine, determined by Chunap (Chagas urine nanoparticle test), are correlated with parasitemia levels in T. cruzi/HIV co-infected patients. METHODOLOGY/PRINCIPAL FINDINGS: T. cruzi antigens in urine of HIV patients (N = 55: 31 T. cruzi infected and 24 T. cruzi serology negative) were concentrated using hydrogel particles and quantified by Western Blot and a calibration curve. Reactivation of Chagas disease was defined by the observation of parasites in blood by microscopy. Parasitemia levels in patients with serology positive for Chagas disease were classified as follows: High parasitemia or reactivation of Chagas disease (detectable parasitemia by microscopy), moderate parasitemia (undetectable by microscopy but detectable by qPCR), and negative parasitemia (undetectable by microscopy and qPCR). The percentage of positive results detected by Chunap was: 100% (7/7) in cases of reactivation, 91.7% (11/12) in cases of moderate parasitemia, and 41.7% (5/12) in cases of negative parasitemia. Chunap specificity was found to be 91.7%. Linear regression analysis demonstrated a direct relationship between parasitemia levels and urine T. cruzi antigen concentrations (p<0.001). A cut-off of > 105 pg was chosen to determine patients with reactivation of Chagas disease (7/7). Antigenuria levels were 36.08 times (95% CI: 7.28 to 64.88) higher in patients with CD4+ lymphocyte counts below 200/mL (p = 0.016). No significant differences were found in HIV loads and CD8+ lymphocyte counts. CONCLUSION: Chunap shows potential for early detection of Chagas reactivation. With appropriate adaptation, this diagnostic test can be used to monitor Chagas disease status in T. cruzi/HIV co-infected patients.
Assuntos
Antígenos de Protozoários/urina , Doença de Chagas/diagnóstico , Coinfecção/diagnóstico , Testes Diagnósticos de Rotina/métodos , Infecções por HIV/complicações , Parasitemia/diagnóstico , Trypanosoma cruzi/isolamento & purificação , Adulto , Linfócitos T CD8-Positivos , Estudos de Casos e Controles , Doença de Chagas/complicações , Doença de Chagas/parasitologia , Doença de Chagas/urina , Coinfecção/imunologia , Coinfecção/parasitologia , Coinfecção/urina , Testes Diagnósticos de Rotina/instrumentação , Diagnóstico Precoce , Feminino , Infecções por HIV/urina , Humanos , Masculino , Pessoa de Meia-Idade , Nanopartículas/química , Parasitemia/imunologia , Parasitemia/parasitologia , Parasitemia/urina , Trypanosoma cruzi/genética , Trypanosoma cruzi/imunologia , Adulto JovemRESUMO
Infection, coinfection and type-specific human papillomavirus (HPV) distribution was evaluated in human immunodeficiency virus (HIV)-positive women from paired cervical and urine samples. Paired cervical and urine samples (nâ=â204) were taken from HIV-positive women for identifying HPV-DNA presence by using polymerase chain reaction (PCR) with three generic primer sets (GP5+/6+, MY09/11 and pU1M/2R). HPV-positive samples were typed for six high-risk HPV (HR-HPV) (HPV-16, -18, -31, -33, -45 and -58) and two low-risk (LR-HPV) (HPV-6/11) types. Agreement between paired sample results and diagnostic performance was evaluated. HPV infection prevalence was 70.6% in cervical and 63.2% in urine samples. HPV-16 was the most prevalent HPV type in both types of sample (66.7% in cervical samples and 62.0% in urine) followed by HPV-31(47.2%) in cervical samples and HPV-58 (35.7%) in urine samples. There was 55.4% coinfection (infection by more than one type of HPV) in cervical samples and 40.2% in urine samples. Abnormal Papanicolau smears were observed in 25.3% of the women, presenting significant association with HPV-DNA being identified in urine samples. There was poor agreement of cervical and urine sample results in generic and type-specific detection of HPV. Urine samples provided the best diagnosis when taking cytological findings as reference. In conclusion including urine samples could be a good strategy for ensuring adherence to screening programs aimed at reducing the impact of cervical cancer, since this sample is easy to obtain and showed good diagnostic performance.
Assuntos
Colo do Útero/virologia , Infecções por HIV/virologia , Papillomaviridae/genética , Infecções por Papillomavirus/virologia , Adulto , Idoso , Coinfecção/diagnóstico , Coinfecção/urina , Coinfecção/virologia , Colômbia , DNA Viral/genética , Feminino , Infecções por HIV/diagnóstico , Infecções por HIV/urina , Papillomavirus Humano 16/genética , Papillomavirus Humano 16/isolamento & purificação , Papillomavirus Humano 31/genética , Papillomavirus Humano 31/isolamento & purificação , Humanos , Programas de Rastreamento/métodos , Pessoa de Meia-Idade , Papillomaviridae/classificação , Papillomaviridae/isolamento & purificação , Infecções por Papillomavirus/diagnóstico , Infecções por Papillomavirus/urina , Reação em Cadeia da Polimerase , Reprodutibilidade dos Testes , Neoplasias do Colo do Útero/prevenção & controle , Neoplasias do Colo do Útero/urina , Neoplasias do Colo do Útero/virologia , Esfregaço Vaginal , Adulto JovemRESUMO
Human papillomavirus is a DNA virus that includes 118 genotypes. HPV16 is responsible for 80% of cervical cancer in women. Men are important reservoirs and major transmitters of HPV to their partners. The aim of this study was to detect HPV DNA and to determine the prevalence of HPV types 6, 11, 16, and 18 in urine samples of men infected with HIV-1. This study included 223 patients infected with HIV-1 from the Center of Reference on HIV/AIDS (CRT-SP) and an outpatient clinic of HIV. Urine samples were collected and after DNA extraction real-time PCR was performed for detection of HPV DNA. Positive samples were then tested by conventional PCR using type-specific primers for the four HPV types. A total of 223 men infected with HIV-1 were tested, 81% of whom were on HAART. Four (5.8%) were positive for HPV6, 18 (26.1%) were positive for HPV11, 22 (31.9%) were positive for HPV16 and five (7.2%) were positive for HPV18 by conventional PCR. Twenty (29%) patients had other HPV types and five patients (1.5%) had multiple types. The mean T CD4+cells count was 517 and 441 cells/mm(3) (P = 0.30), in HPV negative and positive men, respectively. The HIV viral load was higher in the HPV negative group than for in the men with HPV (P = 0.0002). A 30.9% prevalence of HPV was found in asymptomatic urine samples of men infected with HIV-1. This study suggests that urine may be a useful specimen for HPV screening.
Assuntos
Alphapapillomavirus/isolamento & purificação , Infecções por HIV/complicações , HIV-1 , Infecções por Papillomavirus/epidemiologia , Infecções por Papillomavirus/urina , Adolescente , Adulto , Alphapapillomavirus/genética , Alphapapillomavirus/metabolismo , Brasil/epidemiologia , DNA Viral/isolamento & purificação , DNA Viral/urina , Infecções por HIV/urina , Infecções por HIV/virologia , Humanos , Masculino , Programas de Rastreamento/métodos , Infecções por Papillomavirus/complicações , Reação em Cadeia da Polimerase , Prevalência , Sensibilidade e Especificidade , Carga ViralRESUMO
Kidney involvement in children with Human Immunodeficiency Virus (HIV) infection is increasing in prevalence in parallel with the longer survival of HIV-infected patients and the side-effects of new antiretroviral drugs. However, there are only a few reports describing renal tubular disorders in HIV+ children. This is a cross-sectional, case series study evaluating kidney disease in 26 Venezuelan HIV-infected children. The study cohort consisted of 15 girls and 11 boys, with a median age of 5.9 years (25-75th percentile: 3.6-7.8), who had been treated with antiretrovirals for 2.8 +/- 0.4 years, Overall, the patients were short for their age and gender (Z-height: -3.1; 25-75th percentile: -4.94 to -1.98), and 15 showed signs of mild to moderate malnutrition. All of the children had a normal estimated glomerular filtration rate (136 +/- 22.6 ml/min/1.73 m2), and glomerular involvement was only observed in one patient with isolated proteinuria. None had nephromegaly. In contrast, tubular disorders were commonly found. Hypercalciuria was detected in 16 of the patients (UCa/Cr = 0.28; 25-75th percentile: 0.17-0.54 mg/mg), with five of these showing crystalluria. Eight children showed hyperchloremia, and three had frank metabolic acidosis. Kidney stones were absent in all, but one boy had bilateral medullary nephrocalcinosis. Conclusion, in Venezuelan children, HIV infection per se, or its specific treatment, was commonly associated with renal tubular dysfunction, especially hypercalciuria and acidosis, potentially leading to nephrocalcinosis and growth impairment. We recommend renal tubular evaluation during the follow-up of children with HIV infection.
Assuntos
Cálcio/urina , Infecções por HIV/complicações , HIV , Hipercalciúria/epidemiologia , Nefropatias/epidemiologia , Criança , Pré-Escolar , Colorimetria , Estudos Transversais , Feminino , Seguimentos , Infecções por HIV/epidemiologia , Infecções por HIV/urina , Humanos , Hipercalciúria/etiologia , Hipercalciúria/urina , Nefropatias/diagnóstico , Nefropatias/etiologia , Masculino , Prevalência , Venezuela/epidemiologiaRESUMO
The objective of this report was to evaluate oral fluid and urine as optional samples for HIV-1 confirmatory test using DAVIH-BLOT system (Laboratorios DAVIH, La Habana, Cuba), for which they were compared with their corresponding serum samples in a group of 125 individuals. In band pattern analysis, predominant antibodies in positives oral fluid and urine samples against p34, p68, gp41, gp120, were no different from their corresponding sera according to the proportion comparison test (p < 0,001). Relative sensitivity and specificity of this system were 100% in oral fluid and 98, 75% and 100% in urine samples respectively. These results support optional use of oral fluid and urine that, with slight modifications in the diagnostic system, can be applied for HIV-1 antibody confirmation testing.
Assuntos
Sorodiagnóstico da AIDS/métodos , Western Blotting/métodos , Anticorpos Anti-HIV/análise , Infecções por HIV/diagnóstico , HIV-1/imunologia , Saliva/imunologia , Urinálise , Especificidade de Anticorpos , Anticorpos Anti-HIV/urina , Antígenos HIV/imunologia , Infecções por HIV/metabolismo , Infecções por HIV/urina , Humanos , Imunoglobulina G/análise , Imunoglobulina G/imunologia , Sensibilidade e EspecificidadeRESUMO
A descriptive and retrospective study was conducted aimed at knowing the behavior and the frequency of the bacterial isolations most frequently found in different biological samples (sputum, blood and urine) from patients with HIV/AIDS diagnosis that were seen at the Outpatient Department or that were admitted at the Hospital of "Pedro Kouri" Institute of Tropical Medicine from 1997 to 2001. It was observed that most of the isolations were obtained from sputum samples and that Pseudomonas aeuroginosa was the most isolated microorganism in all samples. Pseudomonas aeuroginosa proved to be the most isolated microorganism in sputum with 28.5%. The bacteria most commonly isolated in blood was Staphylococcus epidermidis with 48.5%. The uropathogenic microorganism most found in urine was Escherichia coli with 43.5%.
Assuntos
Infecções por HIV/microbiologia , Infecções por HIV/sangue , Infecções por HIV/urina , Humanos , Estudos Retrospectivos , Escarro/microbiologiaRESUMO
In the present investigation 28 serologically positive HIV-1 patients, 16 patients with AIDS (< 200/mm3 CD4+ T lymphocytes) and 12 with HIV-1 (200 to 500/mm3 CD4+ T lymphocytes) were studied. The Control Group consisted of 11 healthy individuals. The occurrence of alterations in the anthropometric parameters were higher in AIDS patients, compared to HIV-1 and controls patients, indicating a greater level of malnutrition. All individuals in present study showed normal plasma vitamin A levels. Contrasting, urinary excretion of vitamin A were higher in the AIDS Group (0.23 +/- 0.20 mumol/l) than in the HIV Group (0.19 +/- 0.12 mumol/l) and considerably higher than in the Control Group (0.06 +/- 0.05 mumol/l). Urinary excretion of TBARS also were higher in AIDS Group (3.34 +/- 2.65 mumol/l) compared to HIV (1.71 +/- 0.74 mumol/l) and Control Group (1.70 +/- 0.75 mumol/l). These results demonstrate a greater level of malnutrition and elevate excretion of vitamin A and SRATB in urine of AIDS patients. Therefore, monitoring of nutritional status, especially in relation to vitamin A is recommended for patients with HIV and AIDS.
Assuntos
Síndrome da Imunodeficiência Adquirida/urina , Infecções por HIV/urina , HIV-1 , Estado Nutricional , Substâncias Reativas com Ácido Tiobarbitúrico/análise , Vitamina A/urina , Síndrome da Imunodeficiência Adquirida/sangue , Adulto , Análise de Variância , Antropometria , Infecções por HIV/sangue , Humanos , Contagem de Linfócitos , Vitamina A/sangueRESUMO
Immunofluorescence and immunoperoxidase test directed against early viral antigens, and DNA-DNA hybridization were compared with viral isolation for their abilities to detect Cytomegalovirus (CMV) in the urine of 89 HIV infected patients. From the 100 urine samples collected, 70 were found positive by at least one method. Considering viral isolation as the "gold standard" technique, immunofluorescence and immunoperoxidase had a sensitivity of 92.3% and 88% respectively, with a specificity in both cases of 95%. DNA-DNA hybridization showed a sensitivity of 90% but with lower (60%) specificity. All of the three assays were effective in detecting CMV from urine and the technical advantage of each is discussed.