Vanda Akico Ueda Fick de Souza

Involvement of the central nervous system in patients with dengue virus infection

The findings of a neurological evaluation in 85 patients with confirmed, acute, dengue virus infection are described. Signs of central nervous system involvement were present in 18 patients (21.2%). The most frequent neurological symptom was mental confusion. The frequency of neurological involvement did not differ between patients with primary and secondary dengue infection, and the prevalence of central nervous system involvement in dengue fever and dengue hemorrhagic fever also did not differ significantly. The presence of CNS involvement did not influence the prognosis of dengue infection. Dengue viral CSF RNA was found in 7 of 13 patients submitted to a spinal tap, the CSF viral load being less than 1000 copies/ml. PCR was negative in serum samples obtained from three patients on the same day as the CSF samples, suggesting that the dengue virus actively enters the CNS and that the presence of the virus in the CNS does not result from passive crossing of the blood-brain barrier.

Use of an immunoglobulin G avidity test to discriminate between primary and secondary dengue virus infections.

ABSTRACT An enzyme-linked immunosorbent assay-based immunoglobulin G (IgG) antibody avidity test was evaluated by using sera from 57 patients with acute dengue infection. Overall, 55 of 57 patients were correctly classified (27 of 27 with primary dengue and 28 of 30 with secondary dengue). We conclude that the IgG avidity test can be useful for differentiating between acute, primary, and secondary dengue infections.

Identification of Primary and Secondary Measles Vaccine Failures by Measurement of Immunoglobulin G Avidity in Measles Cases during the 1997 São Paulo Epidemic

ABSTRACT Despite almost universal use of measles vaccines in recent decades, epidemics of the disease continue to occur. Understanding the role of primary vaccine failure (failure to seroconvert after vaccination) and secondary vaccine failures (waning immunity after seroconversion) in measles epidemics is important for the evaluation of measles control programs in developing countries. After a measles epidemic in São Paulo, Brazil, 159 cases previously confirmed by detection of specific immunoglobulin M (IgM) antibodies were tested for IgG avidity, and a secondary immune response, defined by an IgG avidity index of at least 30%, was established in 30 of 159 (18.9%) patients. Among the 159 patients, 107 (67.3%) had not been vaccinated and 52 (32.7%) had received one or more doses of measles vaccine. Of the 107 unvaccinated patients, 104 (97.2%) showed a primary immune response, defined as an IgG avidity index of less than 30%. Among the 52 patients with documented vaccination, 25 (48.1%) showed a primary immune response and 27 (51.9%) showed a secondary immune response, thereby constituting a secondary vaccine failure. Primary vaccine failure was observed in 13 of 13 patients vaccinated prior to 1 year of age and in 43.5 and 12.5%, respectively, of patients receiving one or two doses after their first birthdays. These results provide evidence that measurement of IgG avidity can be used to distinguish between primary and secondary vaccine failures in vaccinated patients with measles; the method can also be a useful tool for the evaluation of measles control programs.

Human herpesvirus-8 infection and oral shedding in Amerindian and non-amerindian populations in the Brazilian Amazon region

BACKGROUND Human herpesvirus type 8 (HHV-8) is hyperendemic in Amerindian populations, but its modes of transmission are unknown. METHODS Antibodies against either HHV-8 lytic antigen or HHV-8 latency-associated nuclear antigen (LANA) were detected, by immunofluorescence assays, in 339 Amerindians and 181 non-Amerindians from the Brazilian Amazon. Serological markers of oro-fecal (hepatitis A), parenteral (hepatitis B and C), and sexual (herpes simplex virus type 2 and syphilis) transmission were measured by specific ELISAs. Salivary HHV-8 DNA was detected by use of a nested polymerase chain reaction assay and was sequenced. RESULTS Antibodies against either lytic antigen or LANA were detected in 79.1% of Amerindians and in 6.1% of non-Amerindians (adjusted seroprevalence ratio [SR], 12.63 [95% confidence interval {CI}, 7.1-22.4]; P<.0001). HHV-8 seroprevalence increased with age among Amerindians (P(Trend) < .001) and already had high prevalence in childhood but was not sex specific in either population. The 2 populations did not differ in seroprevalence of oro-fecal or parenteral markers, but seroprevalence of markers of sexual transmission was lower among Amerindians. HHV-8 DNA in saliva was detected in 47 (23.7%) of 198 HHV-8 seropositive Amerindians. Detection of HHV-8 DNA decreased with age (P(Trend) < .04) and was more common in men (SR, 2.14 [95% CI, 1.3-3.5]; P=.003). A total of 36 (76.6%) of the 47 saliva HHV-8 DNA samples were sequenced, and all clustered as subtype E. CONCLUSION The data support the hypothesis of early acquisition and horizontal transmission, via saliva, of HHV-8 subtype E in Amerindian populations.

Seroprevalence of antibodies against varicella-zoster virus and response to the varicella vaccine in pediatric renal transplant patients

Abstract:  The severity of varicella‐zoster virus (VZV) in immunocompromised children, especially in those receiving renal transplants, is well known. However, the use of live attenuated virus vaccine in this population is controversial. This study aimed to: (i) assess the immunization status of pediatric renal transplant recipients at our center; (ii) determine the anti‐VZV antibody titers in such patients; (iii) evaluate the response to VZV vaccine in seronegative children and in those who present low antibody titers (defined as <500 mAU/mL).Vaccinated children were monitored for adverse effects for 8 wk after vaccination. Fifty patients with a mean age of 13.7 yr (range, 3–17 yr) were enrolled. In 49, blood samples were collected and antibodies were screened using ELISA. Seropositivity to VZV was found in 43 (88%), and antibody titers were ≥500 mAU/mL in 37 (75.5%). Of the 12 children who were eligible for vaccination and had antibody titers <500 mAU/mL, one developed varicella before vaccination, two did not meet the inclusion criteria, and three parents refused the vaccination. In the six vaccinated children, there were no adverse reactions to the vaccine, and four (66.6%) responded with anti‐VZV titers ≥500 mAU/mL 6–8 wk after vaccination. In conclusion, after renal transplantation, varicella vaccine is safe with a 66% rate of conversion to high antibody titers.

Salivary antibody detection in epidemiological surveys: a pilot study after a mass vaccination campaign against rubella in São Paulo, Brazil

The sensitivity and specificity of salivary rubella antibody detection was investigated using samples collected from 301 children after a mass vaccination campaign in the state of São Paulo, Brazil. Saliva samples were collected by 2 different methods: directly dribbling into a container or using a commercial collecting device. Corresponding finger-prick blood samples were collected on filter paper. Rubella specific immunoglobulin G (IgG) was measured in saliva by antibody capture radioimmunoassay and in blood samples by indirect enzyme-linked immunosorbent assay. The detection of salivary rubella specific IgG showed good correlation with the detection of rubella antibody in the blood samples. For both collecting techniques the predictive value for a positive saliva test was > 99% compared with the results from the blood tests. However, the predictive value for a negative saliva test was only 58.3% for a dribbled sample, compared to 100% for saliva collected using the commercial device. Moreover, collecting saliva by dribbling from children less than 4 years old was difficult. The detection of rubella specific IgG in saliva collected using a commercial device proved to be sensitive and specific in this epidemiological study, encouraging its more widespread application as a means of surveillance after mass vaccination.

Comparative Study of Kaposi's Sarcoma-Associated Herpesvirus Serological Assays Using Clinically and Serologically Defined Reference Standards and Latent Class Analysis

ABSTRACT Accurate determination of infection with Kaposis sarcoma-associated herpesvirus (KSHV) has been hindered by the lack of a “gold standard” for comparison of serological assays used to estimate KSHV prevalence in serosurveys conducted in different settings. We have evaluated the performance of five in-house (developed at University College London [UCL], United Kingdom, and at the virology laboratory of the Instituto de Medicine Tropical [IMT] in Sao Paulo, Brazil) and two commercial (ABI and DIAVIR) serological assays to detect antibodies to latency-associated nuclear antigen (LANA) and to lytic KSHV antigens. We used a variety of serum samples assembled to represent populations likely to be at high, intermediate, and low risk of KSHV infection in Brazil. Composite reference standard panels were prepared based on clinical and serological parameters, against which assay performances were assessed using conventional Bayesian statistics and latent class analysis (LCA). Against the clinical reference standard, in-house immunofluorescence assays to detect anti-LANA antibodies (IFA-LANA) produced at UCL and IMT had similar performances, with sensitivities of 61% (95% confidence interval [CI], 48% to 74%) and 72% (95% CI, 58% to 83%) and specificities of 99% (95% CI, 94% to 100%) and 100% (95% CI, 96% to 100%), respectively, and only the IMT IFA-LANA was included in LCA, together with the IMT IFA-lytic and four enzyme-linked immunosorbent assays (ELISAs). The LCA indicated that the IMT whole-virus ELISA performed best (sensitivity, 87% [95% CI, 81% to 91%]; and specificity, 100% [95% CI, 98% to 100%]), confirming the results obtained with the conventional statistical approach. Commercially available ELISA-based tests yielded the lowest specificities using a spectrum of serum samples. The evaluation of KSHV serological assays is warranted before planning serosurveys in various settings.

Duality of patterns in hepatitis a epidemiology: A study involving two socioeconomically distinct populations in Campinas, São Paulo state, Brazil

To evaluate the prevalence of antibodies against hepatitis A in two socioeconomically distinct populations, 101 and 82 serum samples from high and low socioeconomic groups, respectively, were analysed for the presence of IgG anti-HAV using a commercial ELISA. The prevalence in low socioeconomic level subjects was 95.0%, whereas in high socioeconomic subjects was only 19.6% (p < 0.001). These data show a duality in Brazil: anti-HAV prevalence in low socioeconomic subjects is similar to that of developing countries, while in high socioeconomic subjects, a pattern typical of developed countries is found. The control of this infection in our country is primarily related to the improvement of sanitation, but especially for high socioeconomic level populations, the use of vaccination against hepatitis A is strongly advisable to avoid the occasional appearance of this disease in adults.

Prevalence of herpes simplex type 2 antibodies and a clinical history of herpes in three different populations in Campinas City, Brazil

OBJECTIVES To determine the seroprevalence of herpes simplex virus type 2 (HSV-2) antibodies and the relation between the history of clinical herpes and the presence of type-specific HSV-2 antibodies in three different populations from the city of Campinas City, Brazil. POPULATION AND METHODS One hundred and one college students, 96 patients with sexually transmitted diseases (STD), and 102 women at delivery were interviewed and blood samples were collected. Total HSV (HSV-1 and HSV-2) antibodies were screened by enzyme-linked immunosorbent assay (ELISA) and type-specific HSV-2 antibodies were detected by Western blot assay. RESULTS Herpes simplex virus antibodies were detected in 66.3% of the students, 97.1% of the women at delivery, and 99.0% of the STD patients. Type-specific HSV-2 antibodies were detected in 6.9% of the students, 22.6% of the women at delivery, and in 53.1% of the STD patients. History of genital herpes was reported by none of the students, by one of the women at delivery, and by 11 of 51 (21.6%) STD patients who were HSV-2 seropositive. Four of the 45 (8.9%) seronegative STD patients reported a history of genital herpes. CONCLUSION The prevalence of HSV-2 infection in Campinas City can be significantly affected by the characteristics of the population studied, as was shown in previous studies. The sensitivity of the history of genital herpes was low in the present series, stressing that prophylactic measures for vertical and horizontal transmission of HSV-2 should not be based only on a positive history of genital ulcers.

Evaluation of a Commercial Real-Time PCR Kit for Detection of Dengue Virus in Samples Collected during an Outbreak in Goiania, Central Brazil, in 2005

ABSTRACT In the past 2 decades, dengue has reemerged in Brazil as a significant public health problem. Clinicians demand a diagnostic test with high sensitivity that is applicable during the early symptomatic phase. We aimed to test two distinct molecular methods on samples from suspected dengue cases during an outbreak in Central Brazil. Acute-phase serum specimens from 254 patients suspected of having dengue were collected during 2005 in the city of Goiânia, Central Brazil. Samples were blindly evaluated by real-time and multiplex PCR in addition to routine immunoglobulin M serology and virus culture. Overall, acute dengue was confirmed by serology, multiplex PCR, or virus isolation for 80% of patients (203/254). Another four patients presented real-time PCR-positive results as the unique marker of dengue. Higher real-time PCR positivity levels and viral loads were observed in the early symptomatic phase of disease (≤5 days) than after this period. Multiplex and real-time PCR assays presented a high kappa agreement (0.85). According to multiplex PCR, 60 samples harbored dengue virus type 3 (DEN-3), 4 samples harbored DEN-2, and 1 sample displayed a pattern compatible with a double infection with DEN-2 and -3. The dengue virus real-time kit was found to be practical and adjustable for high throughput, to display the best performance in the early symptomatic phase of dengue cases, and to be valuable for confirming dengue diagnosis in a timely manner.