Prevalence, incidence, and correlates of chlamydia and gonorrhea among young adult injection drug users.

PURPOSE: To measure prevalence, incidence, and correlates of chlamydia and gonorrhea among injection drug users (IDUs). METHODS: Participants (n = 2129; 63% male, 52% white, ages 18-30 years) in five US cities were tested for chlamydia and gonorrhea by urine LCR assay and completed a standardized questionnaire about demographics and recent sexual behavior. Logistic regression identified correlates of prevalent infection; incidence rates were calculated from 6-month follow-up data. RESULTS: Chlamydia prevalence was 5.2% and did not differ by gender. Gonorrhea prevalence was 0.2% among men and 2.0% among women, P < .001. Among men, younger age [OR (95% CI): 0.89 (0.83-0.96)], age at sexual debut [0.91 (0.83-0.99)], and African American race [2.92 (1.53-5.59)] were associated with chlamydia. Among women, age at sexual debut [1.16 (1.02-1.31)] and commercial sex [1.96 (1.03-3.74)] were associated with chlamydia, and with gonorrhea [1.27 (1.04-1.56)] and [5.17 (1.66-16.11)], respectively. At 6 months, the cumulative incidence of chlamydia was 1.7% among men and 4.4% among women, P = .03; no men and 1.3% of women tested positive for gonorrhea, P = .01. IMPLICATIONS: Prevalence and correlates of chlamydia and gonorrhea were similar to other samples, suggesting that screening criteria need not be modified for IDU populations. The number of behavioral correlates identified was limited; perhaps unmeasured sexual-network-level factors play a role in determining sexually transmitted disease (STD) prevalence.

Multiple drug-resistant Chlamydia trachomatis associated with clinical treatment failure.

In vitro susceptibility testing and genotyping were done on urogenital isolates of Chlamydia trachomatis from 3 patients, 2 of whom showed evidence of clinical treatment failure with azithromycin and one of whom was the wife of a patient. All 3 isolates demonstrated multidrug resistance to doxycycline, azithromycin, and ofloxacin at concentrations >4.0 microg/mL. Recurrent disease due to relapsing infection with the same resistant isolate was documented on the basis of identical genotypes of both organisms. This first report of clinically significant multidrug-resistant C. trachomatis causing relapsing or persistent infection may portend an emerging problem to clinicians and public health officials.

Rapid assessment of sexually transmitted diseases in a sentinel population in Thailand: prevalence of chlamydial infection, gonorrhoea, and syphilis among pregnant women--1996.

OBJECTIVE: To determine the prevalence of sexually transmitted diseases (STDs) among pregnant women in Thailand, where case reporting suggests a marked decrease in STDs following a campaign promoting condom use during commercial sex. DESIGN: Cross sectional study of women at their first visit to the study hospitals' antenatal clinics in Chiang Rai (n = 500) and Bangkok (n = 521). METHODS: First catch urine specimens were tested for Chlamydia trachomatis and Neisseria gonorrhoeae using the Amplicor CT/NG polymerase chain reaction assay. Syphilis and HIV serological testing were performed in the study hospitals' laboratories. RESULTS: The prevalence of chlamydial infection was 5.7%, gonorrhoea 0.2%, and syphilis 0.5% (all VDRL or RPR titres were < or = 1:4). The prevalence of HIV infection was 7.1% in Chiang Rai and 2.9% in Bangkok. In a multivariate logistic regression analysis, chlamydial infection was associated with younger age and with higher gestational age at first antenatal clinic visit, but was not associated with marital status, gravidity, city of enrollment, or HIV infection status. CONCLUSIONS: There was a low prevalence of gonorrhoea and syphilis among these pregnant women in Thailand. Chlamydial infection was detected at a higher prevalence, especially among younger women and women registering later for antenatal care. Testing of pregnant women using easily collected urine specimens and a sensitive nucleic acid amplification assay is a feasible method of rapidly assessing chlamydial and gonococcal prevalence.

Detection of Neisseria gonorrhoeae infection by ligase chain reaction testing of urine among adolescent women with and without Chlamydia trachomatis infection.

BACKGROUND AND OBJECTIVES: Culture, the conventional method for detection of Neisseria gonorrhoeae, requires invasive sampling and stringent specimen transport conditions. The recently developed ligase chain reaction test (LCR; Abbott Laboratories; North Chicago, IL) allows noninvasive sampling and stable transport conditions, but has not been evaluated with specimens from adolescent populations. GOAL OF THIS STUDY: To perform a comparative evaluation of a commercial LCR test and culture for the diagnosis of N. gonorrhoeae in adolescent women. STUDY DESIGN: Urine and endocervical swab specimens from 330 teenage women seen in two public health adolescent clinics were tested by LCR and culture. For resolution of discordant results, a polymerase chain reaction (PCR) test was developed that directly amplifies N. gonorrhoeae DNA from urine samples processed for LCR. RESULTS: Thirty-one of 330 (9.4%) cervical specimens were culture-positive for N. gonorrhoeae, and 30 of 330 (9.1%) urine specimens were positive by LCR. After resolution of 13 discordant results, the sensitivity, specificity, and positive and negative predictive values of LCR for urine were 88.2%, 100%, 100%, 98.7%, respectively, and for culture of endocervical specimens were 82.3%, 98.9%, 90.3% and 98%, respectively. CONCLUSIONS: Although more expensive than culture, LCR offers a sensitive means for the detection of N. gonorrhoeae in urine samples and may be useful for this purpose in settings where pelvic examinations are difficult to perform and simultaneous detection of N. gonorrhoeae and Chlamydia trachomatis is advantageous.

Detection of Chlamydia trachomatis cervical infection by urine tests among adolescents clinics.

PURPOSE: To compare urine ligase and polymerase chain reaction (LCR, PCR) tests for diagnosis of Chlamydia trachomatis cervical infection with PCR and nucleic acid probe (GPA) on cervical specimens in adolescents, as well as risk factors for C. trachomatis infection and prevalence of infection at enrollment. METHODS: Urine and cervical specimens were collected from women aged 13-20 years attending adolescent clinics, and interviews were administered. Urine specimens were tested by PCR and LCR, and cervical specimens by GPA and PCR. Prevalence rates of C. trachomatis infection and gonorrhea were compared by demographic, behavioral, and clinical risk factors. RESULTS: Of 415 women tested, 86 (20.7%) were infected with C. trachomatis as indicated by positive cervical PCR results. A higher prevalence of C. trachomatis infection was seen among adolescents who douched monthly or more frequently, or had gonorrhea; prevalence declined from 25.8% in the first 7 months to 16.3% in the last 14 months of the study (p = .017). A statistically significant protective effect for reported condom use was not observed. Sensitivity of urine PCR was 89.5% and specificity was 100% relative to cervical PCR, compared to 84.9% and 99.4% (urine LCR) and 65.4% and 98.0% (cervical GPA). Sensitivity of urine PCR was higher in women with discharge; urine LCR sensitivity was higher in women < 19 years of age. CONCLUSIONS: Polymerase chain reaction and LCR assays on urine specimens were sensitive, specific, and noninvasive tests in this population of adolescents with high C. trachomatis infection prevalence. Chlamydia trachomatis infection was associated with douching monthly or more frequently. Prevalence of infection declined over the period during which the study was conducted.

Increase in endocervical CD4 lymphocytes among women with nonulcerative sexually transmitted diseases.

To assess associations of nonulcerative sexually transmitted diseases (STDs) with human immunodeficiency virus (HIV)-susceptible leukocytes on female genital mucosa, cervicovaginal specimens from 32 HIV-negative STD clinic patients with gonorrhea, chlamydial infection, or trichomoniasis were compared with specimens from 32 clinic patients without these infections. Twenty-eight patients had single infections (15 gonorrhea, 10 chlamydial infection, 3 trichomoniasis), and 4 had dual infections. A saline vaginal wash and saline suspensions of vaginal wall scrapings, ectocervical scrapings, and endocervical brushings were analyzed by flow cytometry. Specimens from the endocervix had the highest proportions of lymphocytes, monocytes, and Langerhans' cells. The median number of endocervical CD4 lymphocytes/10,000 cells was greater among patients with STDs than among those without (476 vs. 245; P < .001). These data suggest that the endocervix may have a particularly important role in heterosexual HIV transmission and that nonulcerative STDs may facilitate HIV transmission by increasing the presence of CD4 lymphocytes at this site.

Screening high-risk adolescent males for Chlamydia trachomatis infection. Obtaining urine specimens in the field.

BACKGROUND AND OBJECTIVES: Reported case data suggest that few men are being tested for Chlamydia trachomatis (CT) infection (female:male reported case ratio is > 5:1) partially because men seek preventive health services less frequently than women and, until recently, obtaining a CT specimen from men required a urethral swab, which has low patient acceptability. A study was conducted in San Diego, CA, to determine whether urine specimens could be obtained from high-risk teen males in the field using a peer teen outreach approach. GOALS: Identify teen males infected with CT and provide treatment and partner management services. STUDY DESIGN: Prevalence survey of 261 teen males and a program cost evaluation. RESULTS: During the 6.5-month study period (Dec 15, 1995 to June 30, 1996) an estimated 1,860 teen males were approached and 261 submitted a urine specimen; 16 (6.1%) were positive by polymerase chain reaction. All positive males were treated with azithromycin, 1 gm, in the field, and 9 female sex partners were treated, 7 of whom were CT positive. The cost per specimen obtained and per CT infection identified was $103 and $1,677, respectively. The annual cost for adding a peer teen outreach service to an existing STD program using existing staff and adding 1.2 full-time equivalents of outreach time is approximately $25,000. CONCLUSION: Peer teen outreach and in-field collection of urine specimens appear to be an acceptable alternative for screening teen males for CT and should be further evaluated in other communities.

Antigenic variation among strains of Chlamydia pneumoniae.

The antigenic profiles of six strains of Chlamydia pneumoniae were analyzed by the microimmunofluorescence test (MIF) and immunoblotting with human serum and murine monoclonal antibody. MIF-derived antibody titers in serum samples from culture-positive patients were four- to eightfold higher against autologous isolate antigen than they were against the prototype antigen strain TW-183. Sera of patients with respiratory illness that were culture negative and complement fixation positive for Chlamydia spp. produced higher titers by MIF against a strain of C. pneumoniae isolated in the area than they did against TW-183. For two of five cases, the criteria for establishing the diagnosis of acute infection were met only with use of the antigen from the local strain; TW-183 was inadequate for this purpose. Immunoblot profiles revealed antigenic differences between strains that varied with the human serologic response; i.e., unique antigens were recognized by the sera of some individuals and not by the sera of others. Using the reactivity of a genus-specific monoclonal antibody against a major outer membrane protein, we found that strain CWL-011, isolated in Atlanta, Ga., may possess a major outer membrane protein with a molecular mass between those of C. trachomatis L2 and other C. pneumoniae strains. These data provide evidence of several new and unique serotypes of C. pneumoniae and suggest that the serologic diagnosis of C. pneumoniae infection may require the use of antigens from more than one strain of this species.

Enzyme immunoassay to determine exposure to Chlamydia pneumoniae (strain TWAR).

Recent studies suggest that a group of Chlamydia strains known as TWAR, which are now proposed to be a new species called Chlamydia pneumoniae, may be a frequent cause of respiratory disease in the United States and many other countries. Current serotesting methods do not allow rapid screening of large numbers of samples to distinguish C. trachomatis exposure from C. pneumoniae exposure. We developed an enzyme immunoassay to decrease cross-reactivity between immunoglobulin G antibodies reactive with C. trachomatis and C. pneumoniae. Elementary bodies of C. trachomatis or C. pneumoniae were treated with a detergent-chelating solution to decrease the reactivity of the common lipopolysaccharide antigens. Sera from four groups of patients, totaling 143 persons, were tested by this assay. The prevalences of titers of greater than or equal to 128 to C. trachomatis and C. pneumoniae, respectively, were as follows: (i) for 23 women seropositive for C. trachomatis by the microimmunofluorescence test, 21 (91%) and 18 (78%); (ii) for 50 adult blood donors, 13 (26%) and 39 (78%); (iii) for 40 sexually transmitted disease clinic patients, 20 (50%) and 32 (80%); (iv) for 30 healthy children 5 to 7 years old, 0 (0%) and 8 (27%). Western blots (immunoblots) of each antigen corroborated the differential reactivity of C. trachomatis-positive, C. pneumoniae-negative and C. trachomatis-negative, C. pneumoniae-positive serum samples. Western blots of serum samples from rabbits immunized with either C. trachomatis or C. pneumoniae elementary bodies revealed at least two protein bands (30 and 80 kilodaltons) which appeared to represent unique C. pneumoniae antigens.

Comparison of serologic screening tests for brucellosis.

The slide agglutination test (SAT), microagglutination test (MAT), and card agglutination test (CAT) were compared with each other, using the tube agglutination test (TAT) as the standard method, by two reference laboratories to determine effectiveness as screening tests for human brucellosis. TAT titers of 1,253 sera tested in both laboratories were compared. In one laboratory, 1,270 sera were tested by the TAT and SAT, while the other laboratory tested 1,261 sera by both methods. Of these sera, 1,155 were tested in one laboratory by the CAT and 187 sera were tested by the MAT. Compared with that of the TAT (greater than or equal to 160 positive), the sensitivities were 97 to 100% (SAT), 90% (CAT), and 88% (MAT). The specificities were 88 to 89% (SAT), 98% (CAT), and 88% (MAT). For populations with a low prevalence of disease, increased specificity offers higher predictive value, so the CAT and MAT are preferable for screening purposes and the choice between tests depends on the number and frequency of tests performed. All sera reactive in the CAT and MAT should be retested with the TAT.

Detection of immunoglobulin M in cerebrospinal fluid from syphilis patients by enzyme-linked immunosorbent assay.

Cerebrospinal fluid (CSF) samples were evaluated in an immunoglobulin M enzyme-linked immunosorbent assay (IgM ELISA) for syphilis with sonic extracts of Treponema pallidum coated on polystyrene plates. The ELISA procedure was reproducible, and T. pallidum antigens were stable., A total of 15 CSF samples from patients with neurosyphilis, 18 CSF samples from patients with syphilis, 12 CSF samples from patients treated for syphilis, and 494 CSF samples from patients with neurologic or other systemic diseases were tested. The IgM ELISA gave reactive results in all of six symptomatic and congenital neurosyphilitic patients and none of nine asymptomatic neurosyphilitic patients. Of 524 CSF samples from nonneurosyphilitic individuals, 513 were nonreactive, resulting in 98% test specificity. The IgM ELISA in CSF should prove to be useful for confirmation of symptomatic neurosyphilis.

Staining intensities in the fluorescent treponemal antibody-absorption (FTA-Abs) test: association with the diagnosis of syphilis.

In 1984 the reporting system for the fluorescent treponemal antibody-absorption (FTA-Abs) test was changed by the Centers for Disease Control (CDC; Atlanta, GA) to eliminate the borderline report. Factors influencing the reliability of the FTA-Abs test results, i.e., sensitivity, specificity, prevalence of syphilis, prescreening of sera with nontreponemal tests, and reproducibility, were considered before the change in the reporting system was recommended and are reported here. The borderline report, when associated with syphilis, was most frequently also associated with the diagnosis of early primary, dark-field-positive, nontreponemal test-nonreactive syphilis. Whereas elimination of the borderline report decreased the sensitivity of the FTA-Abs test as a confirmatory test from 100% to 99.5%, the specificity increased from 82.5% to 88.7%. The 1+ staining intensity had an association of approximately 5% with the diagnosis of syphilis. The changes in the reporting system were designed to assist the clinician in interpreting the results of the FTA-Abs test in those cases that present diagnostic dilemmas.

Four serologic tests for syphilis: results with comparison of selected groups of sera.

The specificity of the fluorescent treponemal antibody-absorbed (FTA-Abs) test was assessed for 17 sera from syphilitic patients that were nonreactive in the Treponema pallidum immobilization (TPI) test but reactive in the FTA-Abs test. Thirty-three other sera from syphilitic patients and 19 sera from nonsyphilitic individuals were also examined by fluorescent treponemal and microhemagglutination Treponema pallidum (MHA-TP) tests and by the enzyme-linked immunosorbent assay (ELISA). Specific absorptions of sera with calf thymus DNA or Treponema pallidum biotype Reiter (Reiter treponemes) were performed. In quantitative immunofluorescence assays (IFA) with antihuman IgG and IgM conjugates, results were similar to those for reactive sera from a control group. Results of both the MHA-TP and ELISA tests supported the specificity of the FTA-Abs test; reactivity in the latter was not removed by specific absorption either with calf thymus DNA or with Reiter treponemes. This evaluation suggests a format for serodiagnosis in cases in which test results are discrepant.

Evaluation of sera from patients with Lyme disease in the fluorescent treponemal antibody-absorption test for syphilis.

To determine whether the cross-reactivity between Treponema pallidum and Borrelia burgdorferi affects the specificity of the fluorescent treponemal antibody-absorption (FTA-Abs) test for syphilis, sera from patients with Lyme disease or syphilis were examined in a quantitative FTA-Abs test. Sera were diluted serially in phosphate-buffered saline, then in sorbent, and were tested with T. pallidum and B. burgdorferi antigens. Nine of 40 sera from patients with known Lyme disease were reactive at the 1:5 dilution with antigen from T. pallidum; only one serum was reactive at the 1:10 dilution. When both antigens were tested, the titer against B. burgdorferi was always higher than that against T. pallidum. Similarly, sera from patients with syphilis showed cross-reactivity with B. burgdorferi. Although reactivity could be absorbed with Treponemal phagedenis (Reiter strain), simultaneous titration with both antigens was easily performed and designated the etiologic agent.

Rheumatoid factor in syphilis.

Immunoglobulin M (IgM) antibodies directed against IgG antibodies (rheumatoid factor [RF]) are known to occur often in patients with syphilis and to interfere with serological tests measuring specific antibodies of the IgM class. In this study we examined the occurrence and specificity of the RF and demonstrated a simple method to detect and eliminate the RF for a specific Treponema pallidum IgM enzyme-linked immunosorbent assay. We measured the occurrence of the RF with a sensitive enzyme-linked immunosorbent assay and found that it increased with the duration of syphilitic disease: 1 of 13 primary syphilis serum specimens, 3 of 13 secondary syphilis serum specimens, and 10 of 27 latent syphilis serum specimens were reactive in this RF test. Those sera containing IgM RF were immunoprecipitated with anti-human gamma chain antibodies and 2% polyethylene glycol until the RF was removed. One serum specimen from a patient in the secondary stage of syphilis and eight serum specimens from patients with latent disease still presented the RF after immunoprecipitation. Removal of the IgG antibodies also improved the sensitivity of the treponemal IgM test, indicating competition of these antibodies for binding sites of the antigen. The enzyme-linked immunosorbent assays for detection of RF and antitreponemal IgM antibodies are performed on the same plate. Theoretically, only sera positive for both tests have to be immunoprecipitated. But our findings indicated an increase in sensitivity of the IgM enzyme-linked immunosorbent assay after removal of IgG antibodies responsible for competition at the binding sites.

Four-step enzyme-linked immunosorbent assay for detection of Treponema pallidum antibody.

Further studies of a four-step enzyme-linked immunosorbent assay procedure to detect Treponema pallidum antibody are described. High-titered antibody, produced in rabbits by intravenous injection of T. pallidum, was used to coat polyvinyl chloride microtiter plates. To these plates a known concentration of T. pallidum was added, followed in successive steps by serial dilutions of human sera and appropriately diluted peroxidase-labeled anti-human immunoglobulin G antibody. O-Phenylenediamine was the substrate. A total of 340 sera were obtained from the DeKalb County Sexually Transmitted Diseases Clinic, Atlanta, Ga., and examined within 3 days of receipt. Ninety-six percent test agreement between the enzyme-linked immunosorbent assay and the fluorescent treponemal antibody absorption-double staining test was obtained. A total of 372 additional sera stored at -20 degrees C were examined. The overall sensitivity of the enzyme-linked immunosorbent assay with sera from patients with various stages of syphilis was 96%. With sera from uninfected individuals, the specificity of the enzyme-linked immunosorbent assay was 95%. No antigen instability was noted with the two antigen preparations used during this evaluation.

Double-conjugate enzyme-linked immunosorbent assay for immunoglobulins G and M against Treponema pallidum.

An enzyme-linked immunosorbent assay (ELISA) for the simultaneous measurement of immunoglobulin G (IgG) and IgM was developed to detect antibodies to Treponema pallidum. Wells of polystyrene microtiter plates were coated with T. pallidum antigen, diluted patient serum was added, and IgG and IgM which bound to the T. pallidum antigen were measured by the simultaneous addition of alkaline phosphatase-labeled anti-human IgG and horseradish peroxidase-labeled anti-human IgM. Bound IgG was detected first, followed by bound IgM. After development of the procedure, 145 categorized sera were evaluated: 60 from individuals without syphilis; 62 from patients with syphilis, including 22 with primary, 20 with secondary, and 20 with latent phases of syphilis; and 23 from patients with rheumatoid arthritis. Of the 60 sera from individuals without syphilis, 100% were nonreactive for IgG antibody and 16% were reactive for IgM. Of the 23 sera from patients with rheumatoid arthritis, 3 were reactive for IgG and 3 were nonreactive for IgM. Of the 62 sera from patients with syphilis, 61 (98%) were reactive for IgG antibody with increased titers as the stage of syphilis increased, whereas IgM reactivity decreased. This enzyme-linked immunosorbent assay appears to be a simple method for the simultaneous measurement of antibodies under equal assay conditions.

Immunofluorescent staining of Treponema in tissues fixed with formalin.

Immunofluorescent examination of formalin-fixed tissue for Treponema pallidum has generally been unsatisfactory because of nonspecific background fluorescence and poor contrast. We examined the process of treating deparaffinized formalin-fixed tissue sections with 1% ammonium hydroxide (NH4OH) to improve fluorescent staining. Treponema pallidum- and Treponema pertenue-infected rabbit testes or human tissue biopsy specimens fixed in 10% buffered formalin and embedded in paraffin were examined. Sections were cut one week to five years after embedment. Tissues were then stained with fluorescein- or rhodamine-labeled human anti- T pallidum globulin for 30 minutes at 37 degrees C. Treponemes were consistently stained and background staining was generally reduced after NH4OH treatment in both fresh and stored tissue. Cutting sections at a thickness of approximately 2 micron was critical to achieve optimal fluorescence.

Fluorescent treponemal antibody absorption double-staining test evaluation.

The fluorescent treponemal antibody absorption (FTA-ABS) double-staining (DS) test has been developed for microscopes equipped with incident illumination, and the procedure offers many advantages over the FTA-ABS test when tests are performed with this equipment. In this study, 346 fresh sera, including 35 from patients with syphilis, were evaluated by the FTA-ABS DS test. Parameters for investigation included two readers, each using a different microscope; a new FTA-ABS DS test reporting system; sera heated at 56 degrees C for 30 min versus unheated sera; and sera retested after at least 2 weeks of freezer storage. Agreement for FTA-ABS DS test readings between the two microscopes was 99%. Between-test agreement for the FTA-ABS test with the conventional reporting system and the FTA-ABS DS test with the new reporting system was 95%. Sensitivity calculations based on reactivity for the 35 syphilis sera were 94% for the FTA-ABS DS test and 91% for the FTA-ABS test. Specificity calculations based on non-reactivity of nonsyphilis sera were 98% for the FTA-ABS DS test and 93% for the FTA-ABS test. Differences in percentages appeared to be related to borderline readings in the FTA-ABS test. For example, if the same reporting system was used for the reference FTA-ABS test, the specificity was 97%. When sera were examined within 48 h, no difference was observed in results obtained with heated and unheated sera. Sera frozen for 2 weeks showed comparable results in the FTA-ABS DS test and the FTA-ABS test. These findings strongly support the recommendation that the FTA-ABS DS test be accepted as a confirmatory test for syphilis. The new reporting system for the FTA-ABS DS test would be advantageous for the reference FTA-ABS procedure.