Genotypic Characterization of Herpes Simplex Virus Type 1 Isolates in Immunocompromised Patients in Rio de Janeiro, Brazil

<div><p>Herpes simplex virus type 1 (HSV-1) is a prevalent human pathogen that causes a variety of diseases, including an increased risk of developing more severe disease in HIV-infected individuals. In Brazil, there is no information about the molecular epidemiology of HSV-1 infection, especially in HIV-infected individuals. The aim of this study was to perform the genotypic characterization of HSV-1 among HIV-infected patients. A total of 214 serum samples from HIV-positive patients without HSV infection symptoms were enrolled in one of two reference hospitals for HIV infection managing in Rio de Janeiro. The gG and gI genes were analyzed by restriction fragment length polymorphism (RFLP) and full nucleotide sequencing of the US8 (1601 bp), UL44 (1996 bp), and UL23 (1244 bp) regions was performed. A total of 38.3% (82/214) and 32.7% (70/214) of the serum samples tested positive for gG and gI genes, respectively. RFLP analysis classified the HSV-1 as belonging to genotype A. Phylogenetic analysis of the Brazilian samples for the US8, UL44, and UL23 regions demonstrated that the nucleotide identity between Brazilian samples was higher than 97% for all genes. No acyclovir mutation was detected in the patients. The shedding of HSV in the serum samples from HIV-positive patients who were asymptomatic for HSV infection was detected in this work. This is the first report of molecular characterization of HSV-1 in Brazilian samples since there is no previous data available in the literature concerning the genotypic classification and stable distribution of Brazilian strains of HSV-1 in Rio de Janeiro, Brazil.</p></div

Longitudinal Study of Hepatitis A Infection by Saliva Sampling: The Kinetics of HAV Markers in Saliva Revealed the Application of Saliva Tests for Hepatitis A Study

<div><p>Despite the increasing numbers of studies investigating hepatitis A diagnostic through saliva, the frequency and the pattern of hepatitis A virus (HAV) markers in this fluid still remains unknown. To address this issue, we carried on a longitudinal study to examine the kinetics of HAV markers in saliva, in comparison with serum samples. The present study followed-up ten patients with acute hepatitis A infection during 180 days post diagnosis (dpd). Total anti-HAV was detected in paired serum and saliva samples until the end of the follow-up, showing a peak titer at 90^th. However, total anti-HAV level was higher in serum than in saliva samples. This HAV marker showed a probability of 100% to be detected in both serum and saliva during 180 dpd. The IgM anti-HAV could be detected in saliva up to 150 dpd, showing the highest frequency at 30^th, when it was detected in all individuals. During the first month of HAV infection, this acute HAV marker showed a detection probability of 100% in paired samples. The detection of IgM anti-HAV in saliva was not dependent on its level in serum, HAV-RNA detection and/or viral load, since no association was found between IgM anti-HAV positivity in saliva and any of these parameter (p>0.05). Most of the patients (80%) were found to contain HAV-RNA in saliva, mainly at early acute phase (30^th day). However, it was possible to demonstrate the HAV RNA presence in paired samples for more than 90 days, even after seroconversion. No significant relationship was observed between salivary HAV-RNA positivity and serum viral load, demonstrating that serum viral load is not predictive of HAV-RNA detection in saliva. Similar viral load was seen in paired samples (on average 10⁴ copies/mL). These data demonstrate that the best diagnostic coverage can be achieved by salivary anti-HAV antibodies and HAV-RNA tests during 30–90 dpd. The long detection and high probability of specific-HAV antibodies positivity in saliva samples make the assessment of salivary antibodies a useful tool for diagnosis and epidemiological studies. The high frequency of HAV-RNA in saliva and the probability of detection of about 50%, during the first 30 dpd, demonstrate that saliva is also useful for molecular investigation of hepatitis A cases, mainly during the early course of infection. Therefore, the collection of saliva may provide a simple, cheap and non-invasive means of diagnosis, epidemiological surveys and monitoring of hepatitis A infection purposes.</p></div

Characteristics and molecular information of HIV/HSV-positive samples.

<p>Characteristics and molecular information of HIV/HSV-positive samples.</p

Primers used for amplification and sequencing of the UL23, UL44, and US8 genes of HSV-1.

<p>*F, forward; R, reverse; bp, base pairs.</p><p>Primers used for amplification and sequencing of the UL23, UL44, and US8 genes of HSV-1.</p

The Bayesian phylogenetic tree constructed by using nucleotide sequence of US8 coding region (1601 bp).

<p>The GenBank accession number is shown for each sequence used. Posterior probabilities are shown at the branch label. Brazilian sequences are noted in red. Color code: Africa—Green, Asia—Blue, Europe—Pink, United States—purple, Brazil—red.</p

The Bayesian phylogenetic tree constructed by using nucleotide sequence of UL44 (1996 bp).

<p>The GenBank accession number is shown for each sequence used. Posterior probabilities are shown at the branch label. Brazilian sequences are noted in red. Color code: Africa—Green, Asia—Blue, Europe—Pink, United States—purple, Brazil—red.</p

Course of hepatitis A in patients followed during 180 days by serum and salivary diagnostic assays: observations for total anti-HAV (A), IgM anti-HAV (C), and viral load (E) and probabilities of positivity for total anti-HAV (B), IgM anti-HAV (D) and HAV-RNA (F).

<p>Solid lines and shaded areas along them represent for saliva the predicted mean values and 95% confidence intervals, whereas dashed lines and respective shaded areas represent for serum results the predicted mean values and 95% confidence intervals. In A-C-E, circles and triangles represent individual observations for saliva and serum, respectively, and in B-D-F, represent the counts of positive tests (value 1) or negative (value 0) for saliva and serum, respectively. Shape sizes (circles and triangles) are proportional to the counts.</p

Individual analysis of the profile of anti-HAV antibodies response and HAV-RNA detection in saliva samples comparing to serum during the follow-up.

<p>Age (years); *IgM and total anti-HAV are shown as DO/CO; IgM anti-HAV DO/CO ≥ 1.0 were considered positive; Total anti-HAV DO/CO ≤ 1.2 were considered positive; ^# HAV-RNA detection are shown as + (positive) or—(negative); ** Patient 10 (p10) was followed up only until 120 days.</p