The prevalence and persistence of human parvovirus B19 infection in thalassemic patients.

Human parvovirus B19 infection was studied in 60 thalassemic patients in Thailand. Seroprevalence, persistence of parvovirus B19 and their genotypes were identified in blood samples. Prevalence of anti-parvovirus B19 IgG and DNA found in thalassemic patients were 38% and 13%, respectively. Anti-parvovirus B19 IgM could be detected in 4% of these positive anti-parvovirus B19 IgG patients. The seroprevalence and parvovirus B19 DNA in patients with a history of blood transfusion were not significantly higher than those without such a history (44% vs. 34% and 20% vs. 9%, respectively). Phylogenetic analysis of NS1 nucleotide sequences of three parvovirus B19 samples revealed that they were parvovirus B19 genotype 1. They showed low genetic diversity from prototype (Au) strain. We concluded that acute and chronic persistent parvovirus B19 infection were found in the thalassemic Thai patients. Chronic persistence of parvovirus B19 infection might play important clinical role in thalassemic patients because of the high prevalence of parvovirus B19 DNA. Blood transfusion had no significant influence to increase the prevalence of parvovirus B19 infection in thalassemic patients.

Surveillance of subtype and genetic variation of the circulating strains of HIV-1 in Thailand.

Two HIV-1 strains, CRF01_AE and subtype B', were reported in Thailand during the early years of the epidemic. Recently, an intersubtype recombination of HIV-1 strain was found in Thailand. Eight-hundred and twenty-eight samples collected during years 1995-2004 from high-risk groups in Bangkok, northern, northeastern, and southern region of Thailand were studied. HIV-1 env nucleotide sequences were used for phylogenetic analysis of the circulating HIV-1 strain. By single HIV-1 region (env) genotyping, CRFO1_AE was found in 97.3% and HIV-1 subtype B was found in 2.7%. A predominance of CRF01_AE was found in all geographic regions. Parallel analysis of the HIV-1 gag and env genes demonstrated that 2.1% and 4.0% of recombinant HIV-1 strains were found using p17 and p24 region sequences, respectively. The recombinant gag gene was also found in one southern isolate. Phylogenetic analysis of HIV-1 isolated from 20 provinces in 2002 suggested the northern and northeastern isolates were more related than the southern isolates which had the lowest genetic diversity of 0.13. The GPGQ V3 loop tip was also present in isolates from all regions. The molecular epidemiological data from this study may be useful for surveillance design as well as targeting prevention efforts. It also provides information regarding new antigenic regions of circulating strains responsible for the HIV-1 epidemic in Thailand.

Trends of HIV seropositivity at Siriraj Hospital: 13 year's observation from 1992-2004.

OBJECTIVE: The aim of the present report was to observe the trend of seroprevalence rates of HIV seropositivity for routine services at Siriraj Hospital for 13 years. MATERIAL AND METHOD: The prevalence rate of HIV seropositivity was analyzed in three groups of subjects: 1) patients who attended the hospital with HIV related diseases; 2) pregnant women at first visit to the antenatal care clinic; 3) emigrating workers who have applied for employment in foreign countries. RESULTS: Of the 13 year-observation, HIV seroprevalence rates in the groups of patients, pregnant women and emigrating workers was 10.6% (95%CI 8.9-12.3%), 2.0% (95%CI 1.8-2.2%) and 0.6% (95%CI 0.4-0.8%), respectively. CONCLUSION: The low prevalence of HIV seropositivity in the group of emigrating workers may be due to self selection, whereas the prevalence in pregnant women, which was rather consistent at about 2.0%, may represent the infection rate in the general population. The seroprevalence rate measured in the group of pregnant women demonstrates that Thailand should increase efforts to confine the spread of HIV infection in the community.

Prevalence of rabies virus and Hantaan virus infections in commensal rodents and shrews trapped in Bangkok.

Five hundred rodents and shrews (Rattus norvegicus: 458, Rattus rattus: 28, Rattus exulans: 5, Mus musculus: 4 and Suncus murine: 5) trapped from the fresh food markets around Bangkok area were investigated for rabies virus and Hantaan virus infections. No rabies viral antigens in the animals' brains were detected by direct immunofluorescence. On the other hand, antibodies to Hantaan virus were demonstrated in the sera of 7 (1.53%) R. norvegicus caught in various markets using a particle agglutination technique. Further determination of the viral genome in rat lung tissue was performed by reverse transcriptase-polymerase chain reaction (RT-PCR) and nested PCR, 3 (0.66%) out of 7 were positive. HindIII and HifI restriction enzyme analyses showed the pattern of the Hantaan virus genome in 2 samples and that of the Seoul virus genome in the other. The results of the present study suggest that rodents from Bangkok's fresh food markets did not carry rabies. Thus, getting rid of rabies in dogs or cats in the Bangkok area may be easier than anticipated because there are no sources of asymptomatic reservoirs. This may result in the low incidence of rabies patients observed in Bangkok. On the contrary, the presence of antibodies and the Hantaan virus genome and Seoul virus genome in R. norvegicus will definitely provide evidence for physicians to be aware of hemorrhagic fever with renal syndrome (HFRS) and other clinical settings of Hantaan/Seoul virus disease in patients with a history of having contact with rats or their excreta.

Seroprevalence of rickettsial infection in commensal rodents and shrews trapped in the Bangkok Metropolitan Area.

Murine typhus and scrub typhus are important human rickettsial diseases in Thailand. Small mammals, including many species of rodents and shrews, serve as the reservoir host of rickettsial diseases. Rickettsia typhi can be transmitted to humans by fleas causing murine typhus, while infection with Orientia tsutsugamushi causing scrub typhus in humans is transmitted by chiggers. The prevalence of rickettsial infection depends on the geographic area. The seroprevalence of antibody to R. typhi and O. tsutsugamushi was studied in commensal rodents and shrews trapped in markets in the Bangkok Metropolitan Area (BMA). R. typhi and O. tsutsugamushi antigen prepared in the yolk sac of embryonated eggs were used to determine the specific antibody in trapped animals' sera by using fluorescein isothiocyanate (FITC)-anti rat immunoglobulins as a second antibody. Antibody to R. typhi was found in 25 (5%) of 500 sera tested and no antibody to O. tsutsugamushi was detected. R. typhi antibody titer ranged from 40-1280 and was found in Rattus norvegicus (4.2%), Rattus rattus (0.4%), Rattus exulans (0.2%), and Mus musculus (0.2%) trapped in 8 of 47 markets in the BMA. R. typhi antibody was commonly found in R. norvegicus. The authors concluded that murine typhus is an important rickettsial disease and R. norvegicus is an important reservoir species of rodents found in markets of the BMA.

Recombinant envelope protein of HIV-1 subtype E as antigen in HIV-1 antibody detection enzyme immunoassay.

In order to develop a reliable and inexpensive serodiagnostic method to be used for anti-HIV antibody detection in Thailand, recombinant envelope (TM or gp41 subunit) protein of HIV-1 subtype E was produced from prokaryotic cell (Escherichia coli) as the source of antigen in enzyme immunoassay (TE diagnostic EIA kit). HIV-1 gp41 subunit of subtype E was successfully expressed in E. coli in the form of polyhistidine-tagged proteins, comprising of rgp41A (601 bases N-terminal half of TM or 25kDa) and rgp41B (560 bases C-terminal half of TM or 24 kDa) by using an expression vector, pBAD/His C. The amount of protein, dilution of sera, and anti-human IgG labeled HRP used in the EIA test optimized by a checker board titration of the protein and seropositive or seronegative sera, were 5.0 microg/ml, 1:300, and 1:4,000, respectively. The blinded test evaluation of TE-diagnostic EIA in 500 seropositive and 500 seronegative sera which have been simultaneously tested by two available commercial kits and compared with our TE diagnostic EIA, gave 99.6% sensitivity and specificity. The other known genetic subtypes sera such as subtype A (n=5), B (n=9), C (n=4) and D (n=5) were also positive with this EIA. The estimated manufacturer cost per test of rgp41 based anti-HIV antibody detection EIA or TE-diagnostic EIA was about 15 baht. This recombinant envelope (gp41 or TM) protein from HIV-1, which can be produced in large quantities without any hazards from growing the virus and has lower cost to produce anti-HIV antibody serological diagnostic kit, should be considered as an HIV screening test in Thailand.