RESUMO
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.
Assuntos
Humor Aquoso , Uveíte/diagnóstico , Corpo Vítreo , Humor Aquoso/microbiologia , Humor Aquoso/parasitologia , Humor Aquoso/virologia , Citomegalovirus/genética , Citomegalovirus/imunologia , DNA Viral/análise , HIV-1 , Herpesvirus Humano 3/genética , Herpesvirus Humano 3/imunologia , Herpesvirus Humano 4 , Humanos , Imunocompetência , Hospedeiro Imunocomprometido , Reação em Cadeia da Polimerase , Simplexvirus/genética , Simplexvirus/imunologia , Toxoplasma , Uveíte/microbiologia , Uveíte/parasitologia , Uveíte/virologia , Corpo Vítreo/microbiologia , Corpo Vítreo/parasitologia , Corpo Vítreo/virologiaRESUMO
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.
Assuntos
Humanos , Humor Aquoso/microbiologia , Humor Aquoso/parasitologia , Humor Aquoso/virologia , Uveíte/diagnóstico , Corpo Vítreo/microbiologia , Corpo Vítreo/parasitologia , Toxoplasma , Uveíte/microbiologia , Uveíte/parasitologia , Uveíte/virologia , Corpo Vítreo/virologia , DNA Viral/análise , Reação em Cadeia da Polimerase , HIV-1 , Hospedeiro Imunocomprometido , Simplexvirus/genética , Simplexvirus/imunologia , Herpesvirus Humano 4 , Herpesvirus Humano 3/genética , Herpesvirus Humano 3/imunologia , Citomegalovirus/genética , Citomegalovirus/imunologia , ImunocompetênciaRESUMO
Ocular toxoplasmosis is one of the most common complications caused by the infection with the parasite Toxoplasma gondii. The risk of developing eye lesions and impaired vision is considered higher in Brazil than other countries. The clinical diagnosis is difficult and the use of sensitive and specific laboratorial methods can aid to the correct diagnosis of this infection. We compared serological methods ELISA and ELFA, and molecular cPCR, Nested PCR and qPCR for the diagnosis of T. gondii infection in groups of patients clinically evaluated with ocular diseases non-toxoplasma related (G1 = 185) and with lesions caused by toxoplasmosis (G2 = 164) in an Ophthalmology clinic in Brazil. Results were compared by the Kappa index, and sensitivity (S), specificity (E), positive predictive value (PPV), and negative (NPV) were calculated. Serologic methods were in agreement with ELISA more sensitive and ELFA more specific to characterize the acute and chronic infections while molecular methods were discrepant where qPCR presented higher sensitivity, however, lower specificity when compared to cPCR and Nested PCR.
Assuntos
Técnicas de Diagnóstico Molecular/métodos , Saúde Pública , Testes Sorológicos/métodos , Toxoplasma/genética , Toxoplasma/isolamento & purificação , Toxoplasmose Ocular/diagnóstico , Uveíte/diagnóstico , Anticorpos Antiprotozoários/sangue , Brasil , DNA de Protozoário/isolamento & purificação , Ensaio de Imunoadsorção Enzimática , Oftalmologia , Valor Preditivo dos Testes , Reação em Cadeia da Polimerase em Tempo Real , Sensibilidade e Especificidade , Toxoplasma/imunologia , Toxoplasmose Ocular/parasitologia , Uveíte/parasitologiaRESUMO
PURPOSE: To evaluate the ability of real-time quantitative PCR (qPCR) for detectingToxoplasma gondii DNA in the peripheral blood and aqueous humor of patients with toxoplasmic active focal necrotizing retinochoroiditis. METHODS: Fifty-five patients with infectious uveitis seen from 2009 to 2013 at the Department of Ophthalmology and Visual Sciences of the Federal University of São Paulo were enrolled in this study. Forty-three patients had toxoplasmic active focal necrotizing retinochoroiditis, and the remaining 12 had non-toxoplasmic infectious uveitis and served as controls. qPCR analysis forT. gondii DNA was performed on the patients' peripheral blood and aqueous humor samples. RESULTS: The qPCR was positive for T. gondii DNA in 37.21% (16/43) of the aqueous humor samples and 2.33% (1/43) of the peripheral blood samples; further, 16.27% (7/43) of the patients had positive results in both their blood and aqueous humor samples. CONCLUSION: qPCR was able to detect T. gondii DNA in patients with toxoplasmic active focal necrotizing retinochoroiditis in the blood as well as the aqueous humor and can help with the diagnosis of the disease.
Assuntos
Humor Aquoso/parasitologia , Coriorretinite/parasitologia , Reação em Cadeia da Polimerase em Tempo Real/métodos , Toxoplasma/genética , Toxoplasmose Ocular/parasitologia , Uveíte/parasitologia , Coriorretinite/sangue , Coriorretinite/diagnóstico , DNA de Protozoário/análise , DNA de Protozoário/sangue , Feminino , Humanos , Masculino , Valor Preditivo dos Testes , Reprodutibilidade dos Testes , Toxoplasmose Ocular/sangue , Toxoplasmose Ocular/diagnóstico , Uveíte/sangueRESUMO
ABSTRACT Purpose: To evaluate the ability of real-time quantitative PCR (qPCR) for detectingToxoplasma gondii DNA in the peripheral blood and aqueous humor of patients with toxoplasmic active focal necrotizing retinochoroiditis. Methods: Fifty-five patients with infectious uveitis seen from 2009 to 2013 at the Department of Ophthalmology and Visual Sciences of the Federal University of São Paulo were enrolled in this study. Forty-three patients had toxoplasmic active focal necrotizing retinochoroiditis, and the remaining 12 had non-toxoplasmic infectious uveitis and served as controls. qPCR analysis forT. gondii DNA was performed on the patients' peripheral blood and aqueous humor samples. Results: The qPCR was positive for T. gondii DNA in 37.21% (16/43) of the aqueous humor samples and 2.33% (1/43) of the peripheral blood samples; further, 16.27% (7/43) of the patients had positive results in both their blood and aqueous humor samples. Conclusion: qPCR was able to detect T. gondii DNA in patients with toxoplasmic active focal necrotizing retinochoroiditis in the blood as well as the aqueous humor and can help with the diagnosis of the disease.
RESUMO Objetivo: Analisar o uso do PCR em tempo real (qPCR) na detecção do DNA do T. gondii no sangue periférico e no humor aquoso de pacientes com lesões de retinocoroidite focal, ativa por toxoplasmose. Métodos: Cinquenta e cinco pacientes com uveite infecciosa foram incluídos neste estudo. Os pacientes foram atendidos entre 2009 a 2013, no Departamento de Oftalmologia e Ciências Visuais da Universidade Federal de São Paulo. Quarenta e três pacientes tiveram o diagnóstico de lesões de retinocoroidite focal, ativa por toxoplasmose e, os outros 12 tiveram o diagnóstico de uveíte infecciosa não toxoplásmica e, por isso foram usados como grupo controle. A técnica de qPCR foi utilizada na detecção de DNA do T. gondii em amostras de sangue periférico e humor aquoso. Resultados: O qPCR foi positivo para o DNA do T. gondii em 37,21% (16/43) das amostras de humor aquoso, 2,33% (1/43) nas amostras de sangue periférico e, 16,27% (7/43) em ambas amostras simultaneamente. Conclusão: O qPCR foi capaz de detectar o DNA do T. gondii em pacientes com lesões de retinocoroidite focal, ativa por Toxoplasmose, no sangue bem como, no humor aquoso, podendo ajudar no diagnostico.
Assuntos
Feminino , Humanos , Masculino , Humor Aquoso/parasitologia , Coriorretinite/parasitologia , Reação em Cadeia da Polimerase em Tempo Real/métodos , Toxoplasma/genética , Toxoplasmose Ocular/parasitologia , Uveíte/parasitologia , Coriorretinite/sangue , Coriorretinite/diagnóstico , DNA de Protozoário/análise , DNA de Protozoário/sangue , Valor Preditivo dos Testes , Reprodutibilidade dos Testes , Toxoplasmose Ocular/sangue , Toxoplasmose Ocular/diagnóstico , Uveíte/sangueRESUMO
BACKGROUND: Toxoplasma gondii is the main culprit in most cases of infectious uveitis, in both acute and recurrent cases of congenital toxoplasmosis and acquired infections. METHODS: The ocular toxoplasmosis was evaluated in patients at the the reference unit in ophthalmology, in Rio Grande do Norte State, determining the risk factors, and the epidemic, serological and clinical profiles. The production of IgM and IgG antibodies to T. gondii was evaluated by microparticle enzyme immunoassay (MEIA). The same patients diagnosed with fundoscopic alterations have been subjected to the fundus photography procedure. RESULTS: Of the 116 patients with positive serology, 66 patients had bilateral ocular damage and 38 patients showed a higher frequency of lesions of type I. The epidemiological investigation showed that direct contact with cats, the consumption of raw or undercooked meat and direct contact with soil are factors not related to ocular toxoplasmosis development. The characterization of the sample was significant for patients aged 31-40 years. CONCLUSIONS: Ocular toxoplasmosis is widely distributed in Natal and other cities in Rio Grande do Norte state, with special relevance for bilateral lesions in 56.9% of the patients assessed, the most frequent being type I with intraocular disposition in the macula.
Assuntos
Toxoplasma/isolamento & purificação , Toxoplasmose Ocular/epidemiologia , Uveíte/parasitologia , Adolescente , Adulto , Idoso , Anticorpos Antiprotozoários/sangue , Brasil/epidemiologia , Feminino , Humanos , Imunoglobulina G/sangue , Imunoglobulina M/sangue , Masculino , Pessoa de Meia-Idade , Fatores de Risco , Estudos Soroepidemiológicos , Toxoplasma/imunologia , Toxoplasmose Ocular/etiologia , Toxoplasmose Ocular/imunologia , Adulto JovemRESUMO
BACKGROUND/AIMS: To determine if patients with inactive chorioretinitis lesions who experience chronic toxoplasmic uveitis test PCR positive for Toxoplasma in their ocular fluids. METHODS: Two patients undergoing long-term anti-toxoplasmic treatment developed chronic uveitis and vitritis. They underwent therapeutic and diagnostic pars plana vitrectomy. Patient specimens were tested for toxoplasmosis by real-time PCR and nested PCR. Patient specimens were also tested for the presence of Toxoplasma antibodies that recognise allelic peptide motifs to determine parasite serotype. RESULTS: Patients tested positive for Toxoplasma by real-time PCR at the B1 gene in the vitreous and aqueous humours of patient 1, but only the vitreous of patient 2. Patients were not parasitemic by real-time PCR in plasma and blood. During surgery, only old hyperpigmented toxoplasmic scars were observed; there was no sign of active retinitis. Multilocus PCR-DNA sequence genotyping at B1, NTS2 and SAG1 loci established that two different non-archetypal Toxoplasma strains had infected patients 1 and 2. A peptide-based serotyping ELISA confirmed the molecular findings. CONCLUSIONS: No active lesions were observed, but both patients possessed sufficient parasite DNA in their vitreous to permit genotyping. Several hypotheses to explain the persistence of the vitritis and anterior uveitis in the absence of active retinitis are discussed.
Assuntos
Anticorpos Antiprotozoários/sangue , Retinite/parasitologia , Toxoplasma/isolamento & purificação , Toxoplasmose Ocular/parasitologia , Uveíte/parasitologia , Corpo Vítreo/parasitologia , Anti-Infecciosos/uso terapêutico , Antiprotozoários/uso terapêutico , Doença Crônica , DNA de Protozoário , Ensaio de Imunoadsorção Enzimática , Feminino , Técnicas de Genotipagem , Glucocorticoides/uso terapêutico , Humanos , Masculino , Pessoa de Meia-Idade , Prednisona/uso terapêutico , Reação em Cadeia da Polimerase em Tempo Real , Retinite/tratamento farmacológico , Retinite/imunologia , Sorotipagem , Toxoplasma/genética , Toxoplasma/imunologia , Toxoplasmose Ocular/tratamento farmacológico , Toxoplasmose Ocular/imunologia , Combinação Trimetoprima e Sulfametoxazol , Uveíte/tratamento farmacológico , Uveíte/imunologia , VitrectomiaRESUMO
Ophthalmic compromise is infrequent in children with congenital Chagas disease. We present 3 patients under 2 months of age, with ocular involvement, all of them referred to the hospital for ophthalmic evaluation of the premature newborn. The ophthalmic finding was bilateral severe vitreitis (posterior uveitis) related to Chagas disease. They received antiparasitic therapy with a good outcome in all cases. Chagas disease must be considered as differential diagnosis of ocular pathology in those countries where the pathology is endemic, and fundoscopic evaluation must be done in those children with the diagnosis, especially those symptomatic and prematurely born.
Assuntos
Doença de Chagas/congênito , Doença de Chagas/complicações , Doenças em Gêmeos/congênito , Doenças em Gêmeos/complicações , Uveíte/parasitologia , Doença de Chagas/diagnóstico , Doença de Chagas/tratamento farmacológico , Doenças em Gêmeos/diagnóstico , Doenças em Gêmeos/tratamento farmacológico , Feminino , Humanos , Lactente , Uveíte/diagnóstico , Uveíte/tratamento farmacológicoRESUMO
El compromiso ocular es una forma de presentación infrecuente en los niños con la enfermedad de Chagas congénita. Se presentan tres pacientes menores de dos meses de edad con compromiso ocular, todos ellos derivados al hospital para control oftalmológico por prematuridad. El diagnóstico oftalmológico fue de vitreítis bilateral intensa (uveítis posterior) asociada a enfermedad de Chagas. Se realizó tratamiento antiparasitario, con buena respuesta en los tres casos. Debe considerarse la enfermedad de Chagas como diagnóstico diferencial de una patología ocular en los lugares donde la enfermedad es endémica y solicitar una evaluación oftalmológica en los niños con diagnóstico de la enfermedad, en especial aquellos sintomáticos y con antecedente de prematuridad.
Ophthalmic compromise is infrequent in children with congenital Chagas disease. We present 3 patients under 2 months of age, with ocular involvement, all of them referred to the hospital for ophthalmic evaluation of the premature newborn. The ophthalmic finding was bilateral severe vitreitis (posterior uveitis) related to Chagas disease. They received antiparasitic therapy with a good outcome in all cases. Chagas disease must be considered as differential diagnosis of ocular pathology in those countries where the pathology is endemic, and fundoscopic evaluation must be done in those children with the diagnosis, especially those symptomatic and prematurely born.
Assuntos
Feminino , Humanos , Lactente , Doença de Chagas/complicações , Doença de Chagas/congênito , Doenças em Gêmeos/complicações , Doenças em Gêmeos/congênito , Uveíte/parasitologia , Doença de Chagas/diagnóstico , Doença de Chagas/tratamento farmacológico , Doenças em Gêmeos/diagnóstico , Doenças em Gêmeos/tratamento farmacológico , Uveíte/diagnóstico , Uveíte/tratamento farmacológicoRESUMO
El compromiso ocular es una forma de presentación infrecuente en los niños con la enfermedad de Chagas congénita. Se presentan tres pacientes menores de dos meses de edad con compromiso ocular, todos ellos derivados al hospital para control oftalmológico por prematuridad. El diagnóstico oftalmológico fue de vitreítis bilateral intensa (uveítis posterior) asociada a enfermedad de Chagas. Se realizó tratamiento antiparasitario, con buena respuesta en los tres casos. Debe considerarse la enfermedad de Chagas como diagnóstico diferencial de una patología ocular en los lugares donde la enfermedad es endémica y solicitar una evaluación oftalmológica en los niños con diagnóstico de la enfermedad, en especial aquellos sintomáticos y con antecedente de prematuridad.(AU)
Ophthalmic compromise is infrequent in children with congenital Chagas disease. We present 3 patients under 2 months of age, with ocular involvement, all of them referred to the hospital for ophthalmic evaluation of the premature newborn. The ophthalmic finding was bilateral severe vitreitis (posterior uveitis) related to Chagas disease. They received antiparasitic therapy with a good outcome in all cases. Chagas disease must be considered as differential diagnosis of ocular pathology in those countries where the pathology is endemic, and fundoscopic evaluation must be done in those children with the diagnosis, especially those symptomatic and prematurely born.(AU)