Prevalence of cytomegalovirus in Thai blood donors by monoclonal staining of blood leukocytes.

Four hundred and forty-one blood and serum samples were collected during August to October 1998 from the blood donors at the blood bank of Rajvithi Hospital, Bangkok, Thailand. Their ages were varied between 18-55 years. All specimens were tested by immunostaining and ELISA methods. Forty-seven specimens (10.66%) gave positive results by immunostaining. Among these, 20 cases were seropositive and 27 cases were seronegative. The age group between 41-50 years had a high percentage of CMV infection as judged by the immunostaining method, more than the other age groups. By ELISA, 231 cases (52.38%) had positive IgG antibody to CMV, 42 cases (9.52%) were IgM antibody positive and 39 cases (8.84%) were positive for both IgG and IgM antibodies. The age groups between 36-40 years had a higher percentage of IgM antibody positives than the other age groups. Since the immunostaining method can detect early CMV infection, screening for the presence of antibodies alone is not enough to rule out CMV infection. Immunostaining along with ELISA detection of antibodies was useful for determining a decrease in CMV infection.

Rapid diagnosis of cytomegalovirus infection in congenital neonates.

Immunostaining was compared with PCR for diagnosis of congenital CMV infection. IgM and IgG antibody assays were also performed in parallel. Immunostaining gave sensitivity and specificity of 60% and 97% respectively. Correlations among immunostaining, PCR and the presence of IgM antibody was reported. Immunostaining can be used for early diagnosis of congenital CMV infection in parallel with detection of IgM antibody.

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.

The significance of HER-2/neu/c-erbB-2 gene amplification in benign and malignant breast disease.

This study was carried out to investigate the amplification of HER-2/neu oncogene in 66 patients with primary breast cancer and 90 samples from benign breast disease (BBD). The amplification of HER-2/neu oncogene in the DNA of paraffin-embedded specimens was determined by differential PCR. Nineteen out of 66 (28.8%) breast cancer patients showed amplification of the gene. No gene amplification was found in benign breast disease. There was no significant correlation of HER-2/neu amplification with, age, menopausal status, the number of positive nodes, tumor size, estrogen receptor, however, amplification of HER-2/neu gene was strongly correlated with nodal status (p = 0.0049). In node positive patients, the incidence of HER-2/neu amplification was high (43%). These findings indicate that the amplification of HER-2/neu gene may be of pathogenetic significance in breast cancer and may have a poor prognosis in node positive breast cancer patients while no gene amplification in benign breast disease suggests that HER-2/neu amplification is a late molecular alteration event in the pathogenesis of breast cancer.

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.

Seroprevalence of human herpesvirus 6 and 7 infections in the Thai population.

Seroprevalence of human herpesvirus 6 (HHV-6) and 7 (HHV-7) was estimated in the Thai population using indirect immunofluorescence assay to determine serum antibodies to HHV-6 and HHV-7. A total of 333 serum samples obtained from umbilical cord blood and venous blood of healthy persons at Siriraj Hospital and Krabi Hospital during 1990-1993 were investigated. Of 73 infants aged 0-1 month, 73% and 78% were found tob e positive for HHV-6 and HHV-7 antibodies, respectively. Antibody to HHV-6 was detected in age groups 2-3 months (38%), 4-5 months (14%), 6 months (44%), 7-11 months (66%), 1-2 year (84%), 3-4 years (82%), 5-9 years (83%), 10-19 years (83%), 20-29 years (80%), 30-39 years (67%), and over 40 years (58%), respectively. This positive rates of HHV-7 antibody in age groups 2-3 months, 4-5 months, 6 months, 7-11 months, 1-2 years, 3-4 years, 5-9 years, 10-19 years, 21-29 years, 30-39 years, and over 40 years were 50%, 21%, 10%, 37%, 47%, 82%, 75%, 72%, 72%, 67%, and 67%, respectively. At 6 months of age as the starting time of infections, 34% (14/41) and 9% (3/41) of infants had presumed primary infections of HHV-6 and HHV-7, respectively. In the follow-up study, 53% (20/38) of children were infected with HHV-6 prior to HHV-7 and only 5% vice versa. Eighty-four percent of children had acquired antibody to HHV-6 by 1-2 years old while 82% of children had acquired antibody to HHV-7 by 3-4 years old. These results suggest that HHV-6 and HHV-7 are prevalent viruses in the Thai population. The infections of both viruses begin at 6 months of age. However, infection of HHV-7 in most children begins later. The data also provided evidence that antigenic distinction between HHV-6 and HHV-7 existed with a limited cross-reactivity in an antibody test. The antibody responses to HHV-6 and HHV-7 occurred independently.

Genome differences among varicella-zoster viruses isolated in Thailand.

The DNAs of 17 isolates of varicella-zoster virus (VZV) obtained from 17 Thai individuals with normal varicella or zoster infections (no underlying diseases) were compared by restriction endonuclease analysis using BglI, PstI, EcoRI, SmaI and BamHI. The DNA of the Japanese strain, Kawaguchi, was also conducted as a reference DNA. All of virus isolates were confirmed for existence of VZV and VZV-DNA by immunofluorescent test and DNA-hybridization, respectively. Almost all of the Thai epidemiologically unrelated isolates and the Kawaguchi strain could be individually differentiated using BglI, PstI, and EcoRI. The other two isolates were identical in restriction profiles even after five endonuclease digestions which SmaI and BamHI were the two more enzymes used, therefore, they could be discriminated totally into 16 strains from overall 17 isolates. These findings demonstrate the strain variation of wild-type varicella-zoster viruses isolated in Thailand.

Treatment of herpes zoster with Clinacanthus nutans (bi phaya yaw) extract.

A randomized, placebo-controlled trial of the efficacy of topical formulation of Clinacanthus nutans (Bi Phaya Yaw) extract was carried out in 51 patients with varicella-zoster virus infection. The study medication was applied five times daily for 7-14 days until the lesions were healed. The number of patients with lesion crusting within 3 days and with lesion healing within 7 days and 10 days were significantly greater in the C. nutans extract-treated group than the placebo group (p < 0.01). Pain scores were reduced more rapidly in the C. nutans extract-treated group than in the placebo group. There were no side effects of the study medication.

Effects of human alpha, beta and gamma interferons on varicella zoster virus in vitro.

The antiviral effects of interferon (IFN) on varicella zoster virus (VZV) and herpes simplex virus (HSV) in vitro were examined. The values for the 50% inhibitory dose (ID50) of IFN-alpha, beta and gamma determined by plaque reduction assay, were 0.813, 0.650 and 13.750 IU/ml, respectively, against VZV and 18.00, 10.38 and 115.0 IU/ml, respectively, against HSV. Thus IFN-alpha and beta were more effective than IFN-gamma against both VZV and HSV and VZV was more sensitive than HSV to the IFNs. Five mutants of VZV which were resistant to acyclovir (ACV), phosphonoacetic acid (PAA) or bromodeoxyuridine (BUDR) were also sensitive to IFN beta, their average ID50 being 1.31 IU/ml. Analysis of virus-specific proteins by the immunofluorescent technique with various antisera showed that IFN had a significant effect before early protein synthesis.

Immunofluorescence, enzyme-linked immunosorbent assay, particle agglutination and western blot for the detection of antibody to human immunodeficiency virus type 1.

Immunofluorescence assay (IFA) has been applied for detection of antibody to human immunodeficiency virus type 1 (HIV-1). To compare the IFA with an enzyme-linked immunosorbent assay (ELISA) and particle agglutination (PA), we examined the antibody response to HIV-1 in 475 sera from AIDS, PGL and ARC patients as well as several risk groups and healthy persons by three methods. The positive results by any methods were confirmed by western blot (WB). The results by all methods were well correlated on the sera from 45 asymptomatic male homosexuals and 70 female prostitutes. There were some false positive results by ELISA in the sera from prisoners and healthy persons. Four sera from drug abusers were positive only by PA and IFA and were negative by ELISA. All were WB-inconclusive. Particle agglutination and IFA results were compared with western blot analysis on 208 ELISA-positive sera. All IFA-strongly positive sera (84%) were positive by western blot. The sera with weakly positive, negative and inconclusive results by IFA (16%) were possibly any of positive, inconclusive or negative by western blot. By PA, 200 of 208 (97%) sera were PA-positive and 1% of these sera were WB-inconclusive while the PA-negative sera were either negative or inconclusive by western blot. These results suggested that PA is a simple and sensitive method for screening of HIV-1 antibody while IFA could be a primary confirmatory test and western blot would then be used for confirming any IFA-negative or inconclusive results.