Performance of cryptococcal antigen lateral flow assay in serum, cerebrospinal fluid, whole blood, and urine in HIV-infected patients with culture-proven cryptococcal meningitis admitted at a Brazilian referral center.

Cryptococcal meningitis is the most common cause of opportunistic meningitis in HIV-infected patients in Brazil and causes unacceptable high mortality rates. In this study, HIV-infected patients with a first episode of culture-proven cryptococcal meningitis in cerebrospinal fluid (CSF) were prospectively included in order to evaluate sensitivity of cryptococcal antigen (CrAg) lateral flow assay (LFA) in serum, CSF, whole blood (fingerstick), and fresh urine. In addition, HIV-infected patients with other neurological confirmed diseases were included in order to evaluate the specificity of CrAg LFA in serum. Twenty patients with cryptococcal meningitis were included and in 19 of them, CrAg LFA in CSF, serum, and whole blood were positive (95% sensitivity). In 18 patients, India ink test was positive in CSF (90% sensitivity), and in 16 cases, CrAg LFA was positive in urine (80% sensitivity). Thirty-six HIV-infected patients with other neurological diseases had negative results of CrAg LFA in serum (100% specificity). In conclusion, CrAg LFA in serum, CSF, and whole blood showed high sensitivity and specificity. Whole blood CrAg LFA seems to be a good and reliable strategy to improve AIDS-related cryptococcal meningitis diagnosis in Brazil.

Performance of cryptococcal antigen lateral flow assay in serum, cerebrospinal fluid, whole blood, and urine in HIV-infected patients with culture-proven cryptococcal meningitis admitted at a Brazilian referral center

Cryptococcal meningitis is the most common cause of opportunistic meningitis in HIV-infected patients in Brazil and causes unacceptable high mortality rates. In this study, HIV-infected patients with a first episode of culture-proven cryptococcal meningitis in cerebrospinal fluid (CSF) were prospectively included in order to evaluate sensitivity of cryptococcal antigen (CrAg) lateral flow assay (LFA) in serum, CSF, whole blood (fingerstick), and fresh urine. In addition, HIV-infected patients with other neurological confirmed diseases were included in order to evaluate the specificity of CrAg LFA in serum. Twenty patients with cryptococcal meningitis were included and in 19 of them, CrAg LFA in CSF, serum, and whole blood were positive (95% sensitivity). In 18 patients, India ink test was positive in CSF (90% sensitivity), and in 16 cases, CrAg LFA was positive in urine (80% sensitivity). Thirty-six HIV-infected patients with other neurological diseases had negative results of CrAg LFA in serum (100% specificity). In conclusion, CrAg LFA in serum, CSF, and whole blood showed high sensitivity and specificity. Whole blood CrAg LFA seems to be a good and reliable strategy to improve AIDS-related cryptococcal meningitis diagnosis in Brazil

Asymptomatic cryptococcal antigen prevalence detected by lateral flow assay in hospitalised HIV-infected patients in São Paulo, Brazil

OBJECTIVE: To determine the prevalence of asymptomatic cryptococcal antigen (CRAG) using lateral flow assay (LFA) in hospitalised HIV-infected patients with CD4 counts <200 cells/ll. METHODS: Hospitalised HIV-infected patients were prospectively recruited at Instituto de Infectologia Emilio Ribas, a tertiary referral hospital to HIV-infected patients serving the S~ao Paulo State, Brazil. All patients were >18 years old without prior cryptococcal meningitis, without clinical suspicion of cryptococcal meningitis, regardless of antiretroviral (ART) status, and with CD4 counts <200 cells/ll. Serum CRAG was tested by LFA in all patients, and whole blood CRAG was tested by LFA in positive cases. RESULTS: We enrolled 163 participants of whom 61% were men. The duration of HIV diagnosis was a median of 8 (range, 1­29) years. 26% were antiretroviral (ART)-na€ive, and 74% were ARTexperienced. The median CD4 cell count was 25 (range, 1­192) cells/ll. Five patients (3.1%; 95%CI, 1.0­7.0%) were asymptomatic CRAG-positive. Positive results cases were cross-verified by performing LFA in whole blood. CONCLUSIONS: 3.1% of HIV-infected inpatients with CD4 <200 cells/ll without symptomatic meningitis had cryptococcal antigenemia in São Paulo, suggesting that routine CRAG screening may be beneficial in similar settings in South America. Our study reveals another targeted population for CRAG screening: hospitalised HIV-infected patients with CD4 <200 cells/ll, regardless of ART status. Whole blood CRAG LFA screening seems to be a simple strategy to prevention of symptomatic meningitis

A Brazilian report using serological and molecular diagnosis to monitoring acute ocular toxoplasmosis.

BACKGROUND: Toxoplasmosis was recently included as a neglected disease by the Center for Disease Control. Ocular toxoplasmosis is one clinical presentation of congenital or acquired infection. The laboratory diagnosis is being used worldwide to support the clinical diagnosis and imaging. The aim of this study was to evaluate the use of serology and molecular methods to monitor acute OT in immunocompetent patients during treatment. METHODS: Five immunocompetent patients were clinically diagnosed with acute OT. The clinical evaluation was performed by ophthalmologic examination using the Early Treatment Diabetic Retinopathy Study, best-corrected visual acuity, slit lamp biomicroscopy, fundoscopic examination with indirect binocular ophthalmoscopy color fundus photography, fluorescein angiography and spectral optical coherence tomography (OCT). Serology were performed by ELISA (IgA, IgM, IgG) and confirmed by ELFA (IgG, IgM). Molecular diagnoses were performed in peripheral blood by cPCR using the Toxoplasma gondii B1 gene as the marker. Follow-up exams were performed on day +15 and day +45. RESULTS: Only five non-immunocompromised male patients completed the follow up and their data were used for analysis. The mean age was 41.2 ± 11.3 years (median: 35; range 31-54 years). All of them were positive for IgG antibodies but with different profiles for IgM and IgA, as well as PCR. For all patients the OCT exam showed active lesions with the inner retinal layers being abnormally hyper-reflective with full-thickness disorganization of the retinal reflective layers, which assumed a blurred reflective appearance and the retina was thickened. CONCLUSIONS: The presence of IgA and IgM confirmed the acute infection and thus was in agreement with the clinical evaluation. Our results show the adopted treatment modified the serological profile of IgM antibodies and the PCR results, but not the IgG and IgA antibodies and that imaging is a good tool to follow-up patients.

Ocular toxoplasmosis with positive polymerase chain reaction in peripheral blood ­ report of two cases, São Paulo State, Brazil / Toxoplasmose ocular com reação em cadeia da polimerase positiva em sangue periférico ­ relato de dois casos, estado de São Paulo, Brasil

Aims: To describe the use of polymerase chain reaction (PCR) in peripheral blood and demonstrate its importance in the clinical follow-up of patients with ocular toxoplasmosis. Case description: Two immunocompetent patients were clinically diagnosed with acute ocular toxoplasmosis. The routine clinical evaluation consisted of fundus examination using binocular indirect ophthalmoscopy, color fundus photography, fluorescein angiography, and spectral domain optical coherence tomography. The serological diagnosis was made by enzyme-linked immunosorbent assay (ELISA) and confirmed by enzyme-linked fluorescent assay (ELFA). The molecular diagnosis was made by PCR in peripheral blood using the B1 gene of Toxoplasma gondii as marker. The younger patient was male, had previous lesion in the right eye, complained of low visual acuity in the left eye and was under treatment. The older patient was male, had retinal detachment, and presented with sudden loss of acuity in the right eye. The fundus examination revealed chorioretinal scar in the left eye. IgG was reactive, IgM was non-reactive, and PCR was positive in the peripheral blood of both patients. New blood samples were collected for serological and molecular monitoring and PCR remained positive in both cases. Six weeks after treatment with oral sulfadiazine and pyrimethamine, the PCR yielded negative results. Conclusion: The results show that T. gondii antigens may be found in peripheral blood during ocular reactivations and that PCR may be a good tool for the follow-up of patients with ocular toxoplasmosis.

Objetivos: Descrever o uso da reação em cadeia da polimerase (PCR) no sangue periférico e demonstrar sua importância no acompanhamento clínico de pacientes com toxoplasmose ocular. Descrição dos casos: Dois pacientes imunocompetentes foram clinicamente diagnosticados com toxoplasmose ocular aguda. Rotineiramente, a avaliação clínica foi feita por fundoscopia com o uso de oftalmoscópio binocular indireto, retinografia colorida, angiografia fluorescente e tomografia de coerência óptica espectral. A sorologia foi realizada por ensaio imunoenzimático (ELISA) e confirmada por ensaio imunoenzimático fluorescente ELFA (IgG, IgM). O diagnóstico molecular foi realizado por PCR em sangue periférico usando o gene B1 de Toxoplasma gondii como marcador. O paciente mais jovem era do sexo masculino, apresentava lesão prévia no olho direito, queixa de baixa acuidade visual no olho esquerdo e estava sob tratamento. O paciente mais velho era do sexo masculino, apresentava descolamento de retina e súbita diminuição de visão no olho direito. A fundoscopia revelou cicatriz coriorretiniana no olho esquerdo. Ambos os pacientes tinham IgG reagente, IgM não reagente e PCR positivo em sangue periférico. Novas amostras de sangue foram coletadas para monitoramento sorológico e molecular e a PCR permaneceu positiva em ambos os casos. Seis semanas após o início do tratamento com sulfadiazina e pirimetamina oral, os resultados do PCR tornaram-se negativos. Conclusões: Os resultados mostram que antígenos de T. gondii podem ser encontrados em sangue periférico durante as reativações oculares e que a PCR parece ser uma boa ferramenta para o acompanhamento de pacientes com toxoplasmose ocular.

Performance of a real time PCR for leishmaniasis diagnosis using a L. (L.) infantum hypothetical protein as target in canine samples.

Visceral leishmaniasis represents an important public health issue in different parts of the world, requiring that measures be put in place to control the spread of the disease worldwide. The canine leishmaniasis diagnosis is not easy based on clinical signs, since dogs may not develop the infection with recognizable signs. Thus, the laboratorial diagnosis is essential to ascertain the incidence and prevalence of canine leishmaniasis especially in areas with major control efforts. Although, the diagnosis can be performed by the use of different approaches, the molecular methods such as PCR have become an indispensable tool for leishmaniases diagnosis. A TaqMan assay for real-time PCR (Linj31-qPCR) was developed to determine the parasite occurrence in clinical cases of leishmaniasis. The assay targets an L. (L.) infantum hypothetical protein region. The specificity of the assay was verified by using Leishmania World Health Organization reference strains including parasites belonging to subgenus L. (Leishmania), subgenus L. (Viannia), other Leishmania species and Trypanosoma cruzi. The sensitivity was verified by using isolates of L. (L.) amazonensis and L. (L.) infantum. The usefulness of the assay for diagnosis was ascertained by testing 277 samples from dogs in regions endemic for visceral and/or cutaneous leishmaniasis and from regions in which leishmaniasis was not endemic in São Paulo State, Brazil. Diagnosis of canine visceral leishmaniasis (CVL) was determined on these animals by conventional PCR and three serological tests. The dog samples were divided into four groups. I, dogs with CVL (n = 101); II, dogs with other diseases and without CVL (n = 97); III, dogs with American cutaneous leishmaniasis (n = 7), and, IV, dogs without CVL (n = 72) from areas where leishmaniasis was not endemic as control group. Results indicated that Linj31-qPCR was able to identify parasites belonging to subgenus L. (Leishmania) with no cross-amplification with other parasite subgenera. The Linj31-qPCR detected Leishmania parasites DNA in 98% of samples from Group I. In conclusion this methodology can be used as routine diagnostic tools to detect parasites from subgenus Leishmania.

Evolution of cytokine profile during the treatment of cerebral toxoplasmosis in HIV-infected patients.

This study was to follow IFN-γ, TNF-α and IL-10 modulation of peripheral blood mononuclear cells (PBMC) from HIV/cerebral toxoplasmosis patients (CT) during specific treatment. The results were compared with two other groups: HIV patients that had CT at least one year before (P/CT) and individuals with chronic toxoplasmosis (CHR). Blood samples (63) collected from three groups were analyzed. CT, 15 patients (3 blood samples collected one day before Toxoplasma gondii treatment; 7 and 15days during the treatment). P/CT, 5 patients (one blood sample collected at least, one year after the treatment). CHR, 13 individuals with chronic toxoplasmosis (one blood sample). Cytokine levels were assessed by ELISA after PBMC stimulation with T. gondii antigen. CT patients had low IFN-γ; discrete increase at 7th and 15th days; and the levels were recovered in cured patients (P/CT). CT patients had high TNF-α in the beginning of the treatment. TNF-α levels decrease during the treatment (7th and 15th) and in those patients who were treated (P/CT). IL-10 levels were almost similar in CT and P/CT groups but low when compared with CHR individuals. The evolution of the infection was correlated to restoration of IFN-γ response and a decrease of the inflammation. The evaluation of the immune response can provide valuable information and better monitoring of patients during specific treatment.

Antígenos excretados/secretados de Toxoplasma gondii: resposta imune celular e humoral na toxoplasmose cerebral e Aids / Excreted/secreted antigens of Toxoplasma gondii: humoral and cellular immune response in cerebral toxoplastmosis and AIDS

O presente estudo avaliou alguns aspectos da resposta imune celular e humoral na co-infecção toxoplasmose cerebral e Aids (TC/Aids). O primeiro passo foi aperfeiçoar a produção de dois antígenos que foram utilizados em todos os experimentos. Um deles foi o antígeno lisado de taquizoítos (ALT) e o outro, um grupamento de antígenos excretados/secretados (ESA) obtidos dos sobrenadantes de culturas de células VERO, sem soro fetal bovino infectadas com taquizoítos. A seguir foram avaliados os níveis de IgG anti-T. gondii no líquido cefalorraquidiano(LCR) de 99 pacientes com TC/Aids. Em ambos os ensaios (ELISA e Western Blotting), ESA desencadeou reatividade nas amostras desses pacientes, o que não foi observado com o ALT. Concomitantemente, foi padronizado a ELISA, empregando ambos os antígenos, para a detecção de subclasses de IgG em 265 amostras de soro e 270 amostras de LCR. Os grupos de soros foram compostos de 58 pacientes TC/Aids; 49 com Aids/outras neuro infecções/positivos para toxoplasmose; 58 com Aids/outras neuroinfecções/negativos para toxoplasmose; 50 indivíduos soropositivos para toxoplasmose e 50 indivíduos sadios. Os de LCR foram compostos conforme a presença ou não de toxoplasmose ativa sendo: 99 pacientes com TC/Aids; 112 com outras neuroinfecções/soropositivos para toxoplasmose e 59 com outras neuroinfecções/sorologia negativa para toxoplasmose. Somente com ESA obtivemos uma diferenciação no perfil de subclasses de IgG, onde níveis de IgG4 só foram detectados nos pacientes com TC/Aids, e a presença de IgG2 predominou nos indivíduos cronicamente infectados. O valor diagnóstico das IgA e IgE na TC/Aids foi também avaliado. Os resultados mostraram que não foi possível correlacionar tais anticorpos com a reativação da infecção. A resposta imune celular na TC/Aids foi parcialmente avaliada estudando-se os níveis de citocinas (IFN-g, TNF-α, IL-10, IL-12 e IL-4) em sobrenadantes de culturas de células...

Anti-Toxoplasma gondii antibodies in pregnant women and their newborn infants in the region of São José do Rio Preto, São Paulo, Brazil.

CONTEXT AND OBJECTIVE: Toxoplasmosis transmission during pregnancy can cause severe sequelae in fetuses and newborns. Maternal antibodies may be indicators of risk or immunity. The aim here was to evaluate seropositivity for anti-Toxoplasma gondii (anti-T. gondii) immunoglobulin M (IgM) and immunoglobulin G (IgG) antibodies and IgG avidity in pregnant women and their newborn infants. DESIGN AND SETTING: Cross-sectional study in a high-risk pregnancy outpatient clinic. METHODS: Serum samples from pregnant women (n = 87) and their respective newborns (n = 87) were evaluated for anti-T. gondii antibodies using indirect immunofluorescence (IIF) (IgM and IgG), enzyme-linked immunosorbent assay (ELISA) (IgG) and an avidity test. RESULTS: Anti-T. gondii antibodies were identified in 64.4% of the serum samples from the mothers and their infants (56/87). Except for two maternal serum samples (2.3%), all others were negative for anti-T. gondii IgM antibodies, using IIF. The results showed that 92.9% of the pregnant women had high IgG avidity indexes (> 30%) and four samples had avidity indexes between 16 and 30%. Two women in the third trimester of pregnancy were positive for anti-T. gondii IgM antibodies; their babies had avidity indexes between 16 and 30%. The avidity indexes of serum from the other 83 newborns were similar to the results from their mothers. CONCLUSIONS: The results showed that 2% of the pregnant women were at risk of T. gondii transmission during the gestational period. These data seem to reflect the real situation of gestational toxoplasmosis in the northwestern region of the state of São Paulo.

Anti-Toxoplasma gondii antibodies in pregnant women and their newborn infants in the region of São José do Rio Preto, São Paulo, Brazil / Anticorpos anti-Toxoplasma gondii em gestantes e seus neonatos na região de São José do Rio Preto, São Paulo, Brazil

CONTEXT AND OBJECTIVE: Toxoplasmosis transmission during pregnancy can cause severe sequelae in fetuses and newborns. Maternal antibodies may be indicators of risk or immunity. The aim here was to evaluate seropositivity for anti-Toxoplasma gondii (anti-T. gondii) immunoglobulin M (IgM) and immunoglobulin G (IgG) antibodies and IgG avidity in pregnant women and their newborn infants. DESIGN AND SETTING: Cross-sectional study in a high-risk pregnancy outpatient clinic. METHODS: Serum samples from pregnant women (n = 87) and their respective newborns (n = 87) were evaluated for anti-T. gondii antibodies using indirect immunofluorescence (IIF) (IgM and IgG), enzyme-linked immunosorbent assay (ELISA) (IgG) and an avidity test. RESULTS: Anti-T. gondii antibodies were identified in 64.4 percent of the serum samples from the mothers and their infants (56/87). Except for two maternal serum samples (2.3 percent), all others were negative for anti-T. gondii IgM antibodies, using IIF. The results showed that 92.9 percent of the pregnant women had high IgG avidity indexes (> 30 percent) and four samples had avidity indexes between 16 and 30 percent. Two women in the third trimester of pregnancy were positive for anti-T. gondii IgM antibodies; their babies had avidity indexes between 16 and 30 percent. The avidity indexes of serum from the other 83 newborns were similar to the results from their mothers. CONCLUSIONS: The results showed that 2 percent of the pregnant women were at risk of T. gondii transmission during the gestational period. These data seem to reflect the real situation of gestational toxoplasmosis in the northwestern region of the state of São Paulo.

CONTEXTO E OBJETIVOS: A toxoplasmose, quando transmitida durante a gestação, pode causar graves sequelas em fetos e neonatos. Anticorpos maternos podem ser indicadores de risco ou de imunidade. O objetivo foi avaliar a positividade dos anticorpos das classes imunoglobulina M (IgM) e imunoglobulina G (IgG) anti-Toxoplasma gondii (anti-T. gondii), bem como a avidez de IgG em gestantes e seus neonatos. TIPO DE ESTUDO E LOCAL: Estudo transversal em ambulatório de gestação de alto risco. MÉTODOS: Anticorpos anti-T. gondii foram avaliados em amostras de soro de gestantes (n = 87) e seus respectivos neonatos (n = 87) com o uso dos métodos imunofluorescência indireta (IFI) (IgM e IgG), ensaio imunoenzimático (ELISA) (IgG) e avidez. RESULTADOS: Anticorpos anti-T. gondii foram identificados em 64,4 por cento das amostras de soro das mães e seus bebês (56/87). Com exceção de duas amostras de soro materno (2,3 por cento), todas as demais foram negativas anticorpos IgM anti-T. gondii determinado pela IFI. Os resultados mostraram que 92,9 por cento das gestantes tinham índices elevados de avidez de IgG (> 30 por cento) e 4 amostras apresentaram índices de avidez entre 16-30 por cento. Duas gestantes no terceiro trimestre da gravidez eram positivas IgM anti-T. gondii; seus bebês apresentaram índices de avidez entre 16 e 30 por cento. Os índices de avidez dos soros dos outros 83 recém-nascidos foram semelhantes àqueles encontrados nas amostras maternas. CONCLUSÕES: Os resultados mostraram que 2 por cento das gestantes estavam sob risco de transmissão de T. gondii durante o período gestacional. Estes dados parecem refletir a real situação da toxoplasmose gestacional na região noroeste do Estado de São Paulo.

Proteínas excretadas/secretadas(ESAs) de Toxoplasma gondii: emprego no diagnóstico sorológico da toxoplasmose cerebral em pacientes com Aids

Apesar dos recentes avanços no diagnóstico molecular e sorológico da toxoplasmose cerebral em pacientes com aids, ainda existem limitações na prática clínica. O tratamento específico é usualmente iniciado através do diagnóstico presuntivo, baseado em achados clínicos e radiológicos. O presente estudo avalia o emprego das proteínas excretadas/secretadas (ESAs) de Toxoplasma gondii no diagnóstico sorológico da toxoplasmose cerebral em pacientes com aids. A escolha do antígeno foi baseada naqueles produzidos por taquizoítos, uma vez que esta forma é responsável pela disseminação da infecção, assim como estimulação da resposta imune celular e humoral. Pelo método de ELISA (ESA-ELISA), o pool de ESAs recuperadas de sobrenadantes de culturas de células VERO infectadas com taquizoítos (cepa RH) 48 horas pós-infecção teve alta especificidade para soros de pacientes com toxoplasmose cerebral. As reações foram comparadas com a ELISA utilizada na sorologia convencional, na qual se emprega como antígeno, um extrato bruto de taquizoítos. Os ensaios foram realizados em 300 amostras de soros divididos em: Grupo I - 106 soros de pacientes com toxoplasmose cerebral e aids (sintomáticos); Grupo II - 99 soros de indivíduos com toxoplasmose crônica (soropositivos), excluindo-se pela suspeita clínica e triagem da Instituição de origem, os soros suspeitos de HIV positivo; e Grupo III - 95 soros de indivíduos sadios sem toxoplasmose (controle). A ELISA convencional não foi capaz de distinguir os Grupos I e II que apresentaram similar reatividade. Em contraste, ESA-ELISA distinguiu soros de pacientes com toxoplasmose cerebral (p<0,05). Estes soros foram três vezes mais reativos do que aqueles de indivíduos soropositivos (médias de 12.6 para 4.2). Os resultados foram confirmados pela concordância com os resultados em western blotting e pela análise estatística. Esses dados sugerem que ...

Proteínas excretadas/secretadas (ESAs) de Toxoplasma gondii: emprego no diagnóstico sorológico da toxoplasmose cerebral em pacientes com Aids / Excreted/secreted proteins (ESAs) of Toxoplasma gondii: use in the sorologic dianosis of cerebral toxoplasmosis in Aids patients

Apesar dos recentes avanços no diagnóstico molecular e sorológico da toxoplasmose cerebral em pacientes com aids, ainda existem limitações na prática clínica. O tratamento específico é usualmente iniciado através do diagnóstico presuntivo, baseado em achados clínicos e radiológicos. O presente estudo avalia o emprego das proteínas excretadas/secretadas (ESAs) de Toxoplasma gondii no diagnóstico sorológico da toxoplasmose cerebral em pacientes com aids. A escolha do antígeno foi baseada naqueles produzidos por taquizoítos, uma vez que esta forma é responsável pela disseminação da infecção, assim como estimulação da resposta imune celular e humoral. Pelo método de ELISA (ESA-ELISA), o pool de ESAs recuperadas de sobrenadantes de culturas de células VERO infectadas com taquizoítos (cepa RH) 48 horas pós-infecção teve alta especificidade para soros de pacientes com toxoplasmose cerebral. As reações foram comparadas com a ELISA utilizada na sorologia convencional, na qual se emprega como antígeno, um extrato bruto de taquizoítos. Os ensaios foram realizados em 300 amostras de soros divididos em: Grupo I - 106 soros de pacientes com toxoplasmose cerebral e aids (sintomáticos); Grupo II - 99 soros de indivíduos com toxoplasmose crônica (soropositivos), excluindo-se pela suspeita clínica e triagem da Instituição de origem, os soros suspeitos de HIV positivo; e Grupo III - 95 soros de indivíduos sadios sem toxoplasmose (controle). A ELISA convencional não foi capaz de distinguir os Grupos I e II que apresentaram similar reatividade. Em contraste, ESA-ELISA distinguiu soros de pacientes com toxoplasmose cerebral (p<0,05). Estes soros foram três vezes mais reativos do que aqueles de indivíduos soropositivos (médias de 12.6 para 4.2). Os resultados foram confirmados pela concordância com os resultados em western blotting e pela análise estatística. Esses dados sugerem que anticorpos anti-ESAs estão preferencialmente presentes em pacientes com a infecção ativa. Desta form...