Evaluation of a new fourth-generation microwell enzyme-linked immunosorbent assay for detection of HIV-1 subtype B and E antibodies.

The recent fourth-generation enzyme-immunoassays have been used to increase the sensitivity for detecting HIV-1 antibodies and reduce the window period of HIV infection. The HIV antigens utilized in those assays were prepared from HIV-1 clade B which is different from HIV-1 subtypes circulating in Thailand. We evaluated 323 HIV-1 seropositives either B or E subtype to determine whether they were detected with the new combined anti-HIV and the p24 Ag assay. Under evaluation we found that this enzyme immunoassay manufactured by Organon Teknika showed the high sensitivity and specificity with a greater delta (delta) value with B than E subtypes samples (+15.29 vs +5.73).

Chlamydia pneumoniae in community-acquired pneumonia.

Chlamydia pneumoniae has been established recently as an important human respiratory pathogen. The aim of this study was to define the prevalence of C. pneumoniae in community-acquired pneumonia. We prospectively investigated adult patients who were treated as inpatients and outpatients. Acute and convalescent serum samples were obtained from each patient. Serological diagnosis of C. pneumoniae infection was determined by enzyme-linked immunosorbent assay (ELISA). Eighty paired sera were tested for C. pneumoniae-specific IgM, IgG and IgA. Twenty-one patients (26.2%) had serological results compatible with acute C. pneumoniae infection. Eighteen (85.7%) of these infected patients were C. pneumoniae-specific IgM positive, three had a seroconversion of IgA and two had a four-fold or greater increase in C. pneumoniae-specific IgG antibody titer. The most common clinical manifestations of community-acquired pneumonia due to C. pneumoniae were fever (100%), cough (100%), chest pain (47.6%) and shortness of breath (42.9%). Physical examination revealed crackle in 85.7 per cent of the cases. These findings suggest that C. pneumoniae is a common cause of community-acquired pneumonia in Thailand.

HIV type 1 subtype E-infected patients with broadened, dual (B/E) V3 loop serology have increased cross-neutralizing antibodies.

The two prevalent subtypes of HIV-1 circulating in Thailand are subtypes E and B. While the most prevalent subtype continues to be E using molecular typing assays, immunologically, a subset of subtype E-infected patients (3.4% in 1997) have binding antibodies to both the E and B V3 loops in a peptide ELISA. To assess the potential function of this dual (B/E) V3 reactivity, plasmas from patients with genetically defined HIV-1 subtype E infection and either E or B/E V3 serotypes were compared for magnitude and breadth of neutralization of seven primary and laboratory-adapted subtype B and E viruses. Dually reactive (B/E) plasmas showed significantly increased cross-neutralizing activity against subtype B viruses (p < 0.001), and increased neutralization of the panel of viruses overall (p < 0.02), as compared to monoreactive E serotype plasmas. While the total envelope binding antibody titers to both subtype B and E envelopes did not differ significantly between the E and B/E plasmas, 67% of B/E plasmas neutralized >50% of the viruses in the panel, and only 14% of E plasmas showed this broadened neutralizing activity. These data suggest that dual (B/E) V3 loop reactivity may be a marker of broader immune recognition of HIV envelope epitopes in subtype E-infected patients. V3 loop antibody, perhaps in conjunction with antibodies to additional epitopes, may play a role in neutralization of virus isolates from Thailand.

HIV-1 suppression during acute scrub-typhus infection.

BACKGROUND: In HIV-1-infected individuals, viral load has been reported to rise transiently if an acute infection with another organism occurs. Our study was prompted by the unexpected finding that HIV-1 copy number fell during an acute infection with Orientia tsutsugamushi, the causative agent of scrub typhus. METHODS: Serial HIV-1 viral load determinations were made in ten Thai adults with scrub typhus, who were not receiving antiretroviral therapy, and in five HIV-1-infected patients who had other infections (four malaria, one leptospirosis), during and after acute infections. Sera from HIV-1-infected patients with scrub typhus and from mice immunised with O. tsutsugamushi were examined for HIV-1-suppressive activity. FINDINGS: Median viral load 3 days after admission was significantly lower in the scrub-typhus group than in patients with other infections (193% vs 376% of day 28 values, p=0.03). In four O. tsutsugamushi-infected patients HIV-1 RNA copy number fell by three-fold or more compared with day 28 values, and HIV-1 copy numbers were below the assay threshold in two patients with scrub typhus. Five of seven HIV-1 isolates from non-typhus patients with CD4 lymphocytes less than 200 cells/microL were syncytia-inducing variants, whereas all ten isolates from O. tsutsugamushi-infected individuals matched by CD4-cell count were non-syncytia inducing (p=0.03). Sera from an HIV-1-negative patient with scrub typhus had potent HIV-1-suppressive activity in vitro. Sera from typhus-infected mice inhibited HIV-1 syncytia formation and bound by immunofluorescence to HIV-1-infected lymphocytes. INTERPRETATION: HIV-1-suppressive factors are produced during some scrub-typhus infection and should be investigated further in the search for novel strategies for the treatment and prevention of AIDS.

Human immunodeficiency virus type 1 test results in patients with malaria and dengue infections.

Six rapid enzyme immunoassays for the detection of HIV antibody were performed on paired sera from 66 patients with malaria and 9 patients with dengue. Kit specificities ranged from 77% to 100%, demonstrating that more data are needed on cross-reactivity with endemic diseases as the use of rapid HIV tests increases.

HIV-2 infection in Thailand.

Neither the seroprevalence of HIV-2 nor the sensitivity of enzyme immunoassays for the detection of antibodies to this retrovirus have been defined in Thailand. We, therefore, Investigated these enigmas using banked sera previously screened for HIV-1 by a test that did not distinguish between HIV-1 and HIV-2. All 1,013 HIV-seroreactive specimens were positive to HIV-1 on retesting, and 740 (73%) were reactive to both HIV-1 and HIV-2. The thirty-six samples that reacted with HIV-2 at a titer of > or = 1:4,096 were further tested to discriminate between HIV-1 and HIV-2 by immunoblot assays incorporating HIV-2 recombinant proteins. One specimen was untypeable, but all others were determined to be HIV-1. Seventy-three percent of sera from Thai HIV-1 infected subjects cross-reacted with HIV-2, but not a single case of HIV-2 infection could be confirmed. The finding suggests low prevalence of HIV-2 infection in Thailand and that current testing for HIV-2 antibody is not necessary in Thai population.

Application of dried blood spot specimens for serologic subtyping of human immunodeficiency virus type 1 in Thailand.

Dried blood spot (DBS) specimens were assessed as an alternative to plasma for human immunodeficiency virus type 1 (HIV-1) serotyping by V3 loop peptide enzyme immunoassay. Nested PCR capable of distinguishing HIV-1 subtypes B and E was used as the reference standard. Ninety-two percent of DBS samples were typeable as either HIV-1 subtype B or E. Serotype results with DBS and plasma were identical for 254 of 257 specimens. A simple DBS collection method provides a convenient alternative for conducting HIV-1 serotype surveillance while retaining sensitivity and specificity.

Nationwide surveillance of HIV-1 prevalence and subtype in young Thai men.

As part of routine surveillance, an HIV-1 serosurvey of 366,074 members of successive cohorts of young Thai men entering service with the Royal Thai Army (RTA) was conducted between November 1989 and November 1995. We analyzed regional and temporal trends in HIV-1 seroprevalence in young men in Thailand and determined the proportion of infections resulting from subtypes E and B in this population in 1992 and 1995. The prevalence in 1992 was compared with that in 1995 by region and demographic group. The HIV-1 subtype was determined in a random sample of HIV-1-positive specimens in 1992 and 1995 using a V3 peptide enzyme immunoassay. From a peak of 3.7% in 1993, overall seroprevalence declined to 3.0% in 1994 and further in 1995 to 2.5%. Between 1992 and 1995, the absolute decrease in seroprevalence was greatest in the upper North (from 12.5% to 5.3%), where the prevalence has been the highest. Overall, 96.9% and 95.9% of typable specimens were determined to be subtype E in 1992 and 1995, respectively. Decline in HIV-1 seroprevalence among young men in Thailand has continued, which suggests that HIV control programs in Thailand may have been successful in decreasing spread of HIV-1. Almost all HIV-1 infections resulted from subtype E.