The role of the Tat gene in the pathogenesis of HIV infection.

The human immunodeficiency virus Tat regulatory protein is essential for virus replication and for the efficient transcription of HIV-1 provirus, and in the pathogenesis of AIDS. The role of the tat gene was investigated in 300 samples. It was found that 71.7% were subtype CRF_01AE, 9.3% were subtype B, while 11.7 and 7.3% of them were cross-reactive and non-typeable, respectively. Moreover the results from peptide ELISA also showed that a low CD4 cell count was related to a low anti-Tat antibody (p < 0.05), which may be due to the progression of HIV-1, which can be found predominantly in AIDS patients. The results of nested PCR showed that the second Tat exon might also play a role in T-cell activation. Reverse transcription polymerase chain reaction (RT-PCR) was used to measure HIV-1 mRNA expression in PBMC. RT-PCR negative results were found mostly in the asymptomatic HIV-seropositive group (88%). HIV-1 mRNA expression was found to correlate with current immunologic status. The differences in Tat protein sequences from DNA sequencing between the patients who had anti-Tat antibody positive and anti-Tat antibody negative, were not significant (p > 0.05). These results suggested that the Tat amino acid sequences were conserved among each group of samples and did not change significantly compared with the consensus sequence in previous studies. Several factors make Tat an attractive target for vaccine design.

Detection of P. vivax antigens in malaria endemic populations of Nepal by ELISA using monoclonal antibodies raised against Thai isolates.

An indirect enzyme linked immunosorbent assay (ELISA) using monoclonal antibody (MAb) originated from the native Thai isolates of P. vivax (McPV1) and the polyclonal antibody (PAb) raised against Nepali isolates of P. vivax was developed for detection of P vivax antigens in red cell lysates. The assay was specific (100%) since it was positive only with P. vivax-infected erythrocytes and was negative when erythrocytes from 40 healthy individuals from malaria non-endemic areas and 40 P. falciparum infected erythrocytes were tested. When the assay was applied to 203 vivax blood samples already proven by microscopic examination collected from Dhanusha district of Nepal, and using the cut-off level of the mean optical density (OD) (0.144) of 40 healthy individuals who had been living in malaria-endemic areas (0.073) + 2 SD (0.016), the assay could detect 189/203 samples, indicating the sensitivity of the test was 93.1% with a detection limit of erythrocytes of 240 parasites/10(6) erythrocytes. In addition, the assay was negative when 40 blood samples with fever of unknown origin, collected from the same malaria-endemic areas, were tested. However, there was a significant correlation between OD values and parasitemia (r=0.649; p=0.018). The results indicate that MAb-PAb indirect ELISA using MAb raised against Thai isolates of P. vivax as the coating antibodies, and polyclonal antibodies raised against local Nepali isolates as the detecting antibody, could detect P. vivax antigens with high degrees of sensitivity and specificity. Furthermore, it seems that the McPV1 MAb raised against Thai isolates of P. vivax could recognize the antigens of Nepali isolates in a wide range of blood samples.

Prevalence of cytomegalovirus, human herpesvirus-6, and Epstein-Barr virus in periodontitis patients and healthy subjects in the Thai population.

Fifty periodontitis patients and 30 healthy patients with oral cavities were selected from the Faculty of Dentistry, Mahidol University, Bangkok, Thailand, from March 2001 to November 2002. Their ages varied between 15 and 70 years. Among the periodontitis patients, specimens were collected from both disease and healthy sites. All samples were evaluated for the presence of CMV, HHV-6, and EBV-1 by nested PCR. Among the periodontitis patients, CMV was found in 34%, of which 8% were at the disease sites, 10% were at the healthy sites, and 16% were from both sites. EBV was not found in this group of the patients, while HHV-6 was found in 4%, at the disease sites only. CMV was found in one (3.3%) healthy control while HHV-6 and EBV-1 were not found. The depth of sample sites, various demographic and baseline characteristics eg sex, age, occupation and root planning were not associated with the presence of these viruses.

Rapid diagnosis of cytomegalovirus in Thai pediatric AIDS patients.

Blood samples were collected from 100 pediatric AIDS patients for the detection of CMV in pp65-bearing leukocytes (PBLs) by immunoperoxidase staining (IP) and PCR. IgM antibody assay was performed to determine the correlation of antigen and antibody. IP and PCR can be used as methods for the early detection of CMV (prior to the presence of IgM antibody). The sensitivity and specificity of IP were 73% and 97% respectively. IP is superior to PCR in several ways: it is very easy to perform, less time consuming, less expensive, and does not require expensive instruments.