Urine as a diagnostic specimen for the detection of Chlamydia trachomatis in Malaysia by ligase chain reaction.

BACKGROUND AND OBJECTIVES: Noninvasive urine screening for Chlamydia trachomatis infections offers a valuable public health tool, which could be of vast importance in chlamydial control programs. The authors evaluated a new DNA amplification method, ligase chain reaction (LCR). GOALS: The goal was to ascertain whether urine testing could be used as screening method to detect C. trachomatis infections in commercial sex workers, patients at sexually transmitted diseases clinic, and asymptomatic patients in Kuala Lumpur, Malaysia. METHODS: First-void urine specimens from 300 men and 300 women were tested by LCR, as well as by a commercially available enzyme immunoassay. The LCR assay amplifies specific sequences within the chlamydial plasmid with ligand-labeled probes, and the resultant amplicons are detected by an automated immunoassay. Specimens with discrepant results were confirmed by another LCR of the specimen that targeted the gene for the major outer membrane protein (OMP1). RESULTS: There were 31 LCR-positive male urine and 37 LCR-positive female urine specimens. The resolved sensitivity and specificity for the LCR of the male urine specimens were 100% and 99.6%, respectively, whereas for female urine specimens, the sensitivity and specificity were 100% and 98.5%, respectively. After resolution of discrepant test results by OMP1 LCR, the prevalence was 10% for men and 11% for women. The urine enzyme immunoassay was not useful in diagnosing C. trachomatis infections in either men or women, as the resolved sensitivities were 10% and 15.2%, respectively. The specificities were 99.6% for men and 98.9% for women. CONCLUSIONS: Testing first-void urine specimens by LCR is a highly sensitive and specific method to diagnose C. trachomatis infections in men and women, providing health care workers and public health officials with a new molecular amplification assay that uses noninvasive urine specimens for population-based screening purposes.

Imaging of human T-lymphotropic virus type I-associated chronic progressive myeloneuropathies.

We studied magnetic resonance imaging (MRI) of the head and cervical spine and CT of the head in 46 patients (14 men, 32 women) with chronic progressive myeloneuropathy. The findings were correlated with human T-lymphotropic virus type I (HTLV-I) serology, race, country of origin, and age. We found a female predominance of 2:1. Most patients were aged between 30 and 50 years, and most were Caribbean immigrants and black. There were 9 men and 17 women with blood antibody titers to HTLV-I and 7 men and 15 women with cerebrospinal fluid (CSF) titers. All patients with virus or antibodies in blood or CSF were Caribbean immigrants or black. T2-weighted cranial MRI showed scattered areas of high signal intensity in the cerebral white matter, usually in the periventricular and subcortical areas, but not in the posterior cranial fossa. Cranial CT revealed periventricular low density areas, ventricular enlargement, and atrophy MRI of the cervical spine showed atrophy of the cord. Myelography was normal in all 15 patients examined. No imaging differences were observed between the HTLV-I-positive and -negative patients. These findings, although consistent with demyelination, are not specific.

The prevalence of antibody to HTLV-I/II in United States plasma donors and in United States and French hemophiliacs.

Antibody to HTLV-I/II was detected in 19 (0.3%) of 6286 plasma donors from five regions of the United States (US). This seroprevalence rate is approximately 10 times that in whole blood donors. The regional distribution of infection was as follows: Southwest, 0.68 percent; Southeast, 0.45 percent; Midwest, 0.28 percent; Northwest, 0.1 percent; and Northeast, 0.0 percent. Rates of HTLV-I/II infection in blacks (0.74%) and Hispanics (0.66%) were higher (both, p less than 0.001) than those in whites (0.08%). All 19 infected units were donated by subjects aged 30 or older, even though 52.9 percent of the donations came from persons less than 30 years old. Equal rates of HTLV-I/II infection were found in men (0.31%) and women (0.29%). No HTLV-I/II antibody was detected in 154 French and 25 US hemophiliacs who were transfused regularly with noninactivated plasma or its derivatives. This suggests that the transfusion of HTLV-I/II-seropositive plasma products does not transmit the viral infection.

A clinical, hematologic, and immunologic analysis of 21 HTLV-II-infected intravenous drug users.

Infection with human T-cell leukemia virus type II (HTLV-II) has been associated with rare chronic T-cell malignancies and has recently been demonstrated in a significant proportion of American intravenous drug abusers (IVDA). Identification of an HTLV-II-infected cohort of IVDA has allowed analysis of the HTLV-II carrier state. We analyzed clinical, hematologic, and immunologic parameters in 21 HTLV-II-infected IVDA, two HTLV-I-infected IVDA, and 20 uninfected control IVDA identified by serologic screening and by analysis of peripheral blood mononuclear cell (PBMC) DNA by polymerase chain reaction (PCR). An elevated absolute lymphocyte count was observed in 4 of 21 HTLV-II-infected IVDA, 1 of 2 HTLV-I-infected IVDA, and 1 of 20 control IVDA. CD8+ T-cell elevation was observed in three of four HTLV-II IVDA with lymphocytosis and one of two HTLV-I-infected IVDA. Activation of CD8+ T cells in HTLV-II-infected IVDA was suggested by an overall increase in CD8+/HLA-DR+ lymphocytes. Cell fractionation and analysis by PCR of HTLV-II-infected carrier blood showed high levels of HTLV-II provirus in unfractionated PBMC and purified T cells and little or no detectable HTLV-II DNA in B cells or monocytes, indicating that T cells were the most likely target of infection in vivo. The frequency of HTLV-II-infected cells was estimated at approximately 1 in 500 cells or less using dilution analysis by PCR of PBMC DNA. Most HTLV-II-infected IVDA are asymptomatic and have no overt hematologic or immunologic abnormalities, although some manifest benign lymphocytosis.

Transfusion-transmitted HTLV-I/II infection in patients undergoing open-heart surgery.

Sera from 5,244 blood donations collected between 1979 and 1987 were screened for antibody to HTLV-I with an enzyme immunoassay (EIA) whose result was confirmed with a two-step procedure including Western blot (WB) and radio immunoprecipitation. Of 20 repeatedly reactive samples, two were confirmed positive for HTLV-I infection. These blood units were transfused to patients undergoing cardiac surgery. Both recipients of the confirmed anti-HTLV-I positive units were infected with HTLV-I as evidenced by antibody seroconversion. In contrast, six recipients of EIA positive, WB indeterminate blood and nine recipients WB negative blood were not infected with HTLV-I. These results confirm a low prevalence of HTLV-I infection in US blood donors, the capacity of infected units to transmit the virus to recipients, and the importance of an appropriate confirmatory assay.

Screening for anti-human T-lymphotropic virus antibody in blood donors and polytransfused patients in Apulia (Italy).

A stringent procedure for the diagnosis of human T-lymphotropic virus (HTLV) infection was applied to 1,732 volunteer blood donors, 401 patients with various hematological disorders and 78 individuals at high risk for HIV infection. It consisted of a viral lysate-based screening assay (Abbott Laboratories, North Chicago, Ill., USA), and two confirmatory assays (Western blot and radioimmunoprecipitation assay). A confirmed positive sample had to react with at least two different HTLV gene products. Evidence of HTLV infection was not found in either blood donors or patients with hematological disorders. In fact, HTLV infection was only observed in 10 intravenous drug abusers or their sexual partners. Contrary to previous reports that claimed HTLV seroprevalences of between 0.3 and 8% in blood donors from Apulia (Italy), our data suggest that infection with this virus is principally restricted to intravenous drug abusers.

Antibody to human T-lymphotropic virus type I/II (HTLV-I/II) among male inmates entering Maryland prisons.

Serum specimens that had been obtained for routine operational procedures from consecutive entering male inmates during Spring 1987 and the same calendar period in 1988 were tested to identify prevalence and risk groups for antibody to HTLV-I/II. Specimens were assayed for antibody to HTLV-I/II using ELISA, RIPA, and Western blot techniques. Demographics were compared by serostatus using chi 2 and Fisher's exact tests. Of the 1,932 inmates entering prison, 49.3% were 25 years of age or older, 70.1% were black, 62.4% were committed from the Baltimore metropolitan area, 34.1% were intravenous drug users, and 7.0% demonstrated antibody to HIV-1. Among 1,932 inmates, 18 (0.9%) were HTLV-I/II seropositive. All seropositives were black; age greater than 25 years old was significantly (p less than 0.01) associated with seropositivity. Reactivity to HTLV-I/II did not vary significantly by year of entry, HIV-1 serostatus, jurisdiction, offense category, or sentence. Prevalence of HTLV-I/II among incoming male inmates was elevated compared to available local population comparisons. Additional blinded epidemiological serosurveys of antibody to HTLV-I/II are indicated for prison populations in order to monitor the extent and scope of infection in this population.

Monoclonal antibodies to HBsAg: a study of their specificities for eight different HBsAg subtypes.

Spleens from high responder Biozzi mice immunized with purified HBsAg of adw2, adw4 and ayw3 subtypes respectively were fused with N5-1 mouse myeloma cells. Over 30 clones with anti-alpha activity were obtained (principally from fusion adw2) as well as numerous clones with restrictive anti-HBs reactivity. The aim of this study was to characterize 9 different monoclonal antibodies with restrictive reactivity in comparison to a monoclonal anti-alpha antibody. Two different techniques were used: immunodiffusion and enzymoimmunoassay. Already known specificities or common reactivity between some subtypes were found, as w4, subdivision of ayw3 subtype, antigenic determinant common to w4 and adr, absence of the same determinant in adw2 and ayw1, the 2 subtypes having the highest capacity to absorb anti-alpha. Other monoclonal antibodies indicated the presence of previously unidentified subtype specificities: one of them recognized all HBsAg with d determinant but also ayw4, another recognized all HBsAg with y determinant but also adr, 2 others recognized determinants present completely or partially (spur formation) in many subtypes, another one recognized ayw4 and adw4 but also adw2. The combination of new tools such as monoclonal antibodies and synthetic peptides should facilitate the analysis of HBsAg.