Molecular and phenotypic characteristics of neurotropic HIV-1 subtype E.

Although HIV-1 subtype E associated with neurological dysfunction is common, the virological characteristics of HIV-1 isolated from the CNS for this subtype have not yet been identified. In this study, paired blood and CSF isolated from patients with AIDs-defining illnesses were cultured, sequenced and aligned. Phylogenetic tree and nucleotide-distances from both blood and CSF were investigated. Cytopathicity and co-receptor usage of paired blood and CSF isolates were compared to define the specific characteristics of CNS isolates. The results confirmed that CSF isolates showed less cytopathicity. It was found that both blood and CSF isolates used either CXCR4 or CXCR4 and CCR5 as co-receptors. Interestingly, one CSF isolate using CCR3 as a co-receptor was identified. By sequence analysis, the pair-wise distances of envelope gp 120 sequence and those of all variable regions (except V3 region) between blood and CSF isolates were significantly different. The genetic distances in V1/V2 regions of CSF isolates showed more diversity than those of blood isolates. These findings suggest that the evolution of V1/V2 regions of CSF isolates seems to be an advantage for HIV-1 in CNS infection. In contrast, the genetic distance in V4 and V5 regions of CSF isolates showed less diversity, suggesting that conservation in these regions might be necessary during the process of HIV-1 CNS infection.

Clostridium difficile infections in HIV-positive patients.

The prevalence of Clostridium difficile infections in HIV-positive patients with regard to the presence of its enterotoxin was investigated. Enzyme immunoassay (EIA, Meridian Diagnostic Inc) was used for the detection of C. difficile enterotoxin in stool specimens collected from 201 HIV-positive and 271 HIV-negative diarrheal patients. Culture was performed on cycloserine cefoxitin fructose agar. Chromosomal DNA types of C. difficile isolates were determined by pulsed-field gel electrophoresis (PFGE). In the HIV-positive group, C. difficile enterotoxin was found in 58.8% and 12.6% of diarrheal and non-diarrheal patients, repectively, whereas this toxin was found in 36.5% of HIV-negative-diarrheal patients. However, 13.6% of stool samples were negative by toxin assay, but were positive for C. difficile by culture and latex agglutination test. Among 11 isolates from both HIV-positive and HIV-negative patients, 6 patterns of PFGE type were observed: A, B, C, D, E and F.

Incidence of new Salmonella serovar (S. ratchaburi) in Thailand.

Eighteen strains of Salmonella group E from stool samples were confirmed as Salmonella new serovar. 3, 10 : Z35 : 1, 6 by Centre International des Salmonella, Institut Pasteur, Paris, WHO Collaborating Center for Salmonella, Atlanta, USA and Salmonella-Zentrale Hygienischen Institut, Hamburg, Germany. The name of this new serovar was proposed as S. ratchaburi according to the place of its first isolation in Ratchaburi province. The new serovar of Salmonella was sensitive to many antimicrobial agents except streptomycin and erythromycin.

Prevalences of human herpesvirus 6 and human herpesvirus 7 in normal Thai population.

Prevalences of human herpesvirus 6 (HHV-6) and human herpesvirus 7 (HHV-7) DNA were investigated in normal Thai population. Peripheral blood mononuclear cells (PBMC) and saliva were collected from 238 healthy adults in five provinces which might be a representative of each part of the country, and 120 normal children in one province. Prevalences of HHV-6 DNA PBMC were 45.5-74.3% in adults and 78.3% in children, and in saliva, very low prevalences were detected; 5.7-8.6% in adults and 15.0% in children, respectively. Additionally, all HHV-6 DNA detected in this study were variant B. Comparingly to those of HHV-7 DNA, the prevalences were significantly higher than those of HHV-6, ie, 82.9-91.4% in PBMC of adults, 85% in PBMC of children, 84.8-89.0% in saliva of adults and 92.5% in saliva of children. HHV-6 and HHV-7 isolation from saliva specimens were also performed. No HHV-6 could be isolated from any samples, whereas, in the present study, HHV-7 could be isolated as 90.0% from children and as 20.0-54.5% from adults.

Diagnosis of perinatal HIV-1 infection by in-house PCR.

A study on how to apply PCR as a diagnostic test for the infants born to HIV-1 infected mothers is described. All steps including clinical care, blood sampling, specimen processing and PCR analysis were carried out using native facilities and personnel. An open cohort of 130 children was evaluated at birth, 1, 6, 9, 15, and 18 months of age. Definite infection status was assessed by clinical and serological data during an 18 months of follow up period. PCR results were reported as positive or negative when at least 2 concordant data were denoted. This in-house PCR, compared to known infection status, gave 100% sensitivity and 94.4% specificity within 6 months after birth. On the other hand, clinical diagnosis could identify only the infected infants at 9 months of age. The HIV-1 transmission rate from mother to infant was 23.2%. Though this PCR was not at an optimal level of specificity, it was still beneficial to identify uninfected infants in the first year of their lives and avoid unnecessary medical care. Here, we report an in-house PCR that offers good performance at low cost for the diagnosis of HIV-1 vertical transmission.

The latency rate of human herpesvirus 6 (HHV6) in positive and negative human immunodeficiency virus (HIV) infection of intravenous drug users (IVDU).

The seropositive and latency rates of HHV6 among IVDU with positive and negative HIV and control group were demonstrated. By immunofluorescent antibody test, no differences in the seropositive rates were found among these three groups. All groups had seropositive rate at the average 89% and GMT antibody 1:26. This meant that most of them had previous infection with HHV6. In addition, HHV6-DNA was determined and classified into subgroups: HHV6A and HHV6B, by polymerase chain reaction. The prevalence of HHV6-DNA indicated HHV6 latency in vivo. High latency rate of HHV6 was found in all three groups (the average 54%). Moreover, HHV6B (49%) had a higher frequency than HHV6A (5%); HHV6a was found only in IVDU with or without HIV infection. The result suggested that the HHV6 latency in IVDU with positive HIV may possibly transactivate HIV. The pathogenesis of HHV6 in AIDS patients should be further investigated. However, this research finding is useful for treatment, health care, prevention and control of AIDS in case of dual infections and latency of herpesvirus infection in AIDS.