Evaluation of a novel solid-phase immunoassay, Clearview Chlamydia, for the rapid detection of Chlamydia trachomatis.

Clearview Chlamydia (Unipath Limited, Bedford, United Kingdom) is a rapid immunoassay for the direct detection of Chlamydia trachomatis antigen. This assay was evaluated against the tissue culture method by using 376 paired endocervical specimens. The Clearview assay had a sensitivity of 93.5% and a specificity of 99% when it was compared with the tissue culture method. This assay does not require specialized equipment or trained personnel and yields results within 30 min from the time that a specimen is collected.     

Evaluation of chlamydiazyme for the detection of genital infections caused by Chlamydia trachomatis.

Chlamydiazyme is a 4-h enzyme-linked immunoassay that detects an antigen of Chlamydia trachomatis directly in clinical specimens. This immunoassay was compared with cell culture for the diagnosis of chlamydial infections of the genital tract. The assay was evaluated at five clinics with a total of 1,277 cervical specimens of which 239 were culture positive. At three of these clinics where urethral samples were taken from males, 99 of 363 samples were culture positive. The sensitivity of the assay averaged 89.5% for detecting cervical infections and 78.8% for detecting male urethral infections. Specificity was 97.0% when samples from either males or females were tested. Some patients who were culture negative were infected with chlamydiae according to both Chlamydiazyme and a monoclonal antibody test that detected a chlamydial antigen distinct from the antigen detected by Chlamydiazyme. If the 15 females and 2 males who were positive by both immunoassays but culture negative were considered positive for chlamydial infection, the specificity of the assay was 98.4% in females and 97.7% in males. Chlamydiazyme is a simple and relatively rapid immunoassay that has sufficient sensitivity and specificity to supplant culture in the detection of genital chlamydial infections.     

Diagnosis by AMPLICOR PCR of Chlamydia trachomatis infection in urine samples from women and men attending sexually transmitted disease clinics.

Screening of urine specimens from men for Chlamydia trachomatis infection by a commercial PCR assay (AMPLICOR C. trachomatis Test; Roche Diagnostic Systems, Inc., Branchburg, N.J.) is a sensitive and specific noninvasive diagnostic assay. Since screening of women for C. trachomatis infection with the AMPLICOR C. trachomatis Test has been limited to use with endocervical swab specimens, we conducted an evaluation of the AMPLICOR C. trachomatis Test for the detection of C. trachomatis using female urine samples and compared the results of those obtained by in vitro culture and PCR of endocervical swab specimens. For 713 men we compared the performance of AMPLICOR C. trachomatis Test with urine specimens with that of culture of urethral specimens. For specimens that were PCR positive and culture negative, two additional tests were used to resolve the discrepancies: direct fluorescent-antibody assay (DFA) of sediment from a spun endocervical specimen culture vial and major outer membrane protein-based PCR of the sediment from the endocervical specimen culture vial. Of 525 urine specimens from females, 67 (12.8%) were PCR positive, and 41 (7.8%) endocervical specimens from the 525 women were culture positive. After resolution of the discrepancies, the resolved sensitivity of the urine PCR was 93.3%, whereas the sensitivity of endocervical swab specimen culture was 67.3%. Of 468 female endocervical swab specimens, 47 (10.0%) had a positive PCR result and 33 (7.0%) were culture positive. The resolved sensitivity of the endocervical swab specimen PCR was 86%. Of 415 matched female urine and endocervical swab specimens, there were 49 confirmed infections; 30 (61.2%) specimens were positive by culture of the endocervical swab specimen, 40 (81.6%) were positive by confirmed endocervical swab specimen PCR, 43 (87.8%) were positive by confirmed urine PCR, and all 49 (100%) were positive by either endocervical swab specimen PCR or urine PCR. For men, the resolved sensitivity of the urine PCR was 88%, and the sensitivity of culture was only 50.7%. These results indicate that urine PCR is highly sensitive for the detection of C. trachomatis in both women and men and provides a noninvasive technique for routine screening for chlamydial infection.     

Detection of Chlamydia trachomatis in endocervical specimens by polymerase chain reaction.

A rapid and sensitive polymerase chain reaction (PCR)-based assay for detection of Chlamydia trachomatis in cervical specimens is described. This assay consists of (i) sample preparation which avoids the use of heat, centrifugation, or organic extractions; (ii) rapid, two-temperature PCR amplification of C. trachomatis cryptic plasmid sequences; and (iii) capture and colorimetric detection of amplified DNA in microwell plates. PCR was compared with culture by using 503 cervical specimens. After resolution of discrepant specimens with a confirmatory PCR assay directed against the chlamydial major outer membrane protein gene, PCR had a sensitivity of 97% and a specificity of 99.7% while culture had a sensitivity of 85.7% and a specificity of 100%. In a separate study, PCR was compared with a direct specimen enzyme immunoassay (Chlamydiazyme; Abbott Diagnostics) by using 375 cervical specimens. After resolution of discrepant specimens, PCR had a sensitivity and specificity of 100%, while the enzyme immunoassay had a sensitivity of 58.8% and a specificity of 100%.     

Comparison of DNA probe, monoclonal antibody enzyme immunoassay, and cell culture for the detection of Chlamydia trachomatis.

A total of 201 endocervical specimens were obtained from patients with a clinical or epidemiological history suggestive of chlamydial infection. These specimens were tested by DNA probe (Gen-Probe, San Diego, Calif.) and the IDEIA III (Boots-Celltech, Berkshire, United Kingdom) monoclonal antibody enzyme immunoassay and compared with cell culture for detection of Chlamydia trachomatis. Discrepancies between cell culture and antigen detection methods were resolved by direct fluorescent-antibody testing. In a population with a 17.4% prevalence, the sensitivities and specificities of these assays were 82.8 and 99.4%, respectively, for the DNA probe assay and 97.1 and 98.1%, respectively, for the IDEIA III.     

Multicenter evaluation of the AntigEnz Chlamydia enzyme immunoassay for diagnosis of Chlamydia trachomatis genital infection.

To evaluate the AntigEnz Chlamydia enzyme immunoassay (EIA) (Baxter/Bartels, Issaquah, Wash.), we studied 320 men and 1,209 women attending clinics for sexually transmitted diseases in Baltimore, San Francisco, and Seattle. At examination, two separate swabs were obtained from each patient, one for chlamydial culture and one for EIA. Cervical samples were collected from women, and urethral samples were collected from men. The prevalence of chlamydial infection by culture was 9% in Baltimore (n = 532), 11% in Seattle (n = 500), and 9% in San Francisco (n = 497). To resolve specimens with discrepant culture and EIA results, the EIA transport buffer was centrifuged and the resuspended pellet was stained by direct immunofluorescence to determine whether elementary bodies were present. Overall sensitivity of the AntigEnz Chlamydia assay compared with culture was 87% in men and 86% in women, and overall specificities were 94 and 97%, respectively. Differences between centers were seen, with sensitivities ranging from 76% among men and 79% among women in Seattle to 100% among men and 95% among women in Baltimore. With a true positive considered to be either a culture-positive or an EIA- and direct immunofluorescence-positive specimen, the revised sensitivity was 91% in men and 88% in women. Overall revised specificity was 99% in both men and women. We conclude that in this high-prevalence population, the sensitivity and specificity of this assay compare favorably with those of other noncultural antigen detection tests for the diagnosis of chlamydial genital infection.     

Multicenter comparative evaluation of two rapid immunoassay methods for the detection of Chlamydia trachomatis antigen in endocervical specimens

Objective: To evaluate two rapid immunoassay methods, QuickVue-Chlamydia (Quidel Corp., San Diego California) and Kodak SureCell (Kodak Corp., Rochester, NY) for the detection of Chlamydia trachomatis antigen in endocervical swabs from high- and low-risk females.
Methods: Seven hundred and twenty-four females attending three clinics were enrolled in the study. The results were compared to McCoy's or BGMK cell culture and discrepancies resolved with polymerase chain reaction and direct fluorescent antibody tests performed on left-over culture specimens.
Results: The sensitivity, specificity, predictive value of a positive and predictive value of a negative of the QuickVue Chlamydia assay were 92.0%, 99.1%, 92.0% and 99.1%, respectively. The sensitivity, specificity, predictive value of a positive and predictive value of a negative of the SureCell assay were 90.0%, 99.8%, 98.6% and 98.8%, respectively.
Conclusions: The performances of the two immunoassay methods were similar, and slight differences in sensitivity and specificity were not statistically significant. Both immunoassay methods performed well in high- and low-risk patient groups, both for symptomatic and for asymptomatic patients.     

Detection of Chlamydia trachomatis in genital specimens by the Chlamydiazyme test.

Cotton swabs were used to collect two specimens each from 416 patients (206 males, 210 females) attending a sexually transmitted disease clinic. The first swab was transported in Specimen Storage Reagent and extracted in Specimen Dilution Buffer for enzyme immunoassay by Chlamydiazyme (Abbott Laboratories); the second swab was extracted into 2SP and inoculated into McCoy cell cultures. In the first phase of the study (215 patients: 111 males, 114 females) enzyme immunoassay results were positive (optical density greater than or equal to 0.1) in 30 of 35 instances in which Chlamydia trachomatis was isolated (sensitivity, 86%). Of 18 false-positive enzyme immunoassay results, 15 (83%) were cervical swabs (specificity, 90%). In a phase II study, using a modified Chlamydiazyme kit, 201 patients were tested (95 males, 106 females). Of 41 chlamydial isolates, 8 were not detected by the Chlamydiazyme test (sensitivity, 81%). Only three positive Chlamydiazyme test results could not be confirmed by culture (specificity, 98%). Overall, Chlamydiazyme assay provided a rapid (4 h), sensitive, and specific assay for the detection of chlamydial antigens.     

Comparison of the Gen-Probe PACE 2 system, direct fluorescent-antibody, and cell culture for detecting Chlamydia trachomatis in cervical specimens

A chemiluminescent-labeled DNA probe (Gen-Probe PACE 2 System) for detection of Chlamydia trachomatis in endocervical specimens was evaluated. For each specimen, tissue cell culture, direct immunofluorescent staining (DFA), and DNA probe assay were performed. Thirty of 318 (9.4%) specimens were positive for C. trachomatis. The sensitivities, specificities, and positive and negative predictive values of the DNA probe compared with cell culture were 93%, 98%, 85%, and 99%, respectively, and for DFA these same values were 81%, 99%, 83%, and 99%, respectively. The Gen-Probe PACE 2 System is a reliable method for the rapid detection of endocervical chlamydial infection.     

Detection of Chlamydia trachomatis in urine samples by polymerase chain reaction and enzyme immunoassay

First-void urine samples collected from sexually transmitted diseases (STD) clinic patients were examined by a nested polymerase chain reaction (PCR) and a commercial enzyme immunoassay (IDEIA Chlamydia) for the diagnosis of Chlamydia trachomatis urethritis or cervicitis. The primers for the PCR amplified a target in the major outer membrane protein (MOMP) gene in C trachomatis while the IDEIA detected genus-specific chlamydial lipopolysaccharide. Discrepant results were resolved by retesting urine specimens with a second (plasmid-based) PCR and taking urethral or endocervical swab results into consideration. For 231 men (chlamydial prevalence 20.4%), the sensitivity, specificity, positive and negative predictive values were 59.6%, 99.5%, 96.6% and 90.6% for urine IDEIA, 68.1%, 99.5%, 97% and 92.4% for urethral swab IDEIA and 97.9%, 99.5%, 97.9% and 99.5% for urine PCR. The corresponding rates for 66 women (chlamydial prevalence 54.6%) were 19.4%, 100%, 100% and 50.8% for urine IDEIA, 86.1%, 96.7%, 96.9% and 85.3% for endocervical swab IDEIA and 91.7%, 93.3%, 94.3% and 90.3% for urine PCR. Hence, in a high prevalence population, the urine IDEIA was a suitable alternative to the male urethral swab IDEIA but significantly less sensitive than the endocervical swab IDEIA. The urine PCR was, however, much more sensitive than the urine IDEIA for both men and women and could replace the endocervical swab IDEIA for the diagnosis of chlamydial cervicitis.     

Vulval swabs as alternative specimens for ligase chain reaction detection of genital chlamydial infection in women.

A ligase chain reaction (LCR)-based assay was recently shown to be highly sensitive and specific for the detection of Chlamydia trachomatis not only in cervical specimens but also in first-void urine (FVU) specimens form women. The suitability of using vulval swabs as an alternative specimen that can be obtained by noninvasive means for the diagnosis of genital chlamydial infection by LCR was investigated. In a first study of 169 women, vulval, endocervical, and urethral swabs were tested by LCR, culture, and a combination of enzyme immunoassay (EIA) followed by confirmation by direct fluorescent-antibody assay (DFA), and the results were compared with those obtained by testing FVU specimens by LCR and EIA-DFA by using a specimen from an infected patient as a reference standard. Of the 169 women tested, 27 (16%) were shown to be infected. Whereas LCR showed high sensitivities with all specimen types (85.2% for vulval, urine, and endocervical specimens; 92.6% for urethral swabs), the sensitivities of culture and EIA-DFA were high only with endocervical swabs (74.1 and 70.4%, respectively), being 22.2 and 40.7%, respectively, with vulval swabs. In addition, urine testing by EIA-DFA also showed a poor sensitivity (48.1%). In order to further compare LCR performance with vulval specimens to that with FVU specimens, a second study was carried out with specimens from 312 women, of whom 26 were infected. Comparable sensitivity was obtained by LCR with vulval swabs (88.5%; 23 of 26) and FVU specimens (92.3%; 24 of 26). The results indicate that vulval swabs may serve as suitable alternative to specimens that can be obtained by noninvasive means for the detection of C. trachomatis by LCR.     

Comparison of Kodak Surecell Chlamydia Test Kit with culture for the diagnosis of chlamydial conjunctivitis in infants.

The accuracy of the Surecell Chlamydia Test Kit (Kodak Clinical Products, Rochester, N.Y.) in detecting neonatal conjunctival infection caused by Chlamydia trachomatis was determined by comparison of this enzyme immunoassay with the isolation of C. trachomatis in tissue culture. Kodak Surecell is a rapid monoclonal antibody-based membrane capture enzyme immunoassay which can be processed in the office of a physician. The sensitivity and specificity compared to culture in detecting C. trachomatis in conjunctival specimens from infants with conjunctivitis were 93 and 96%, respectively. The test does not require specialized equipment or trained personnel and would be ideal for physicians who see low numbers of infants with possible chlamydial conjunctivitis in their offices.     

Detection of Chlamydia trachomatis in urine specimens from women by ligase chain reaction.

The performance of a plasmid-based ligase chain reaction (LCR) with urine specimens was compared with those of cell culture of cervical swabs and enzyme immunoassay with urine specimens for the detection of Chlamydia trachomatis infection in women who had attended a family planning clinic. The prevalence of chlamydial infection determined by LCR was 3.1%. Discrepant results among the three assays were resolved by testing urine by a second LCR assay based on the C. trachomatis chromosomal gene encoding the major outer membrane protein. Sensitivity, specificity, and positive and negative predictive values for the cell cultures were 56.3, 100, 100, and 98.4%, respectively, whereas those for the enzyme immunoassay were 18.8, 100, 100, and 97.1%, respectively, and those for LCR were 87.5, 100, 100, and 99.5%, respectively. LCR thus provides a highly sensitive and specific noninvasive screening method for detecting genital chlamydial infections in women.     

Evaluation of enzyme immunoassay (Chlamydiazyme) for detecting Chlamydia trachomatis in genital tract specimens.

An enzyme immunoassay (Chlamydiazyme) for detecting Chlamydia trachomatis was evaluated on genital specimens from 96 men and 272 women attending a clinic for sexually transmitted diseases (STD clinic). Compared with a direct immunofluorescence test for chlamydial elementary bodies, the enzyme immunoassay had a sensitivity of 58% on specimens from men, a specificity of 90%, a positive predictive value of 93%, and a negative predictive value of 88%; the assay had a sensitivity of 67% on specimens from women, a specificity of 89%, a positive predictive value of 63% and a negative predictive value of 90%. Immunofluorescence provided the most stringent test for the performance of the enzyme immunoassay as values were improved a little when a cell culture procedure was used for comparison. Further evidence for the lack of sensitivity was the detection of elementary bodies, sometimes in large numbers, in the enzyme immunoassay buffer of 13 of 19 specimens that had given a negative enzyme immunoassay result and the finding in comparative titrations of four laboratory strains that the enzyme immunoassay was at least 100-fold less able to detect chlamydiae than either immunofluorescence or the cell culture procedure. Lack of specificity may be associated with the finding that the enzyme immunoassay antibody reacted with strains of Acinetobacter calcoaceticus, Escherichia coli, Gardnerella vaginalis, Neisseria gonorrhoeae and group B streptococci. The enzyme immunoassay was not considered to be sufficiently sensitive, specific, or reproducible for routine use.     

Evaluation of a new optical immunoassay for diagnosis of neonatal chlamydial conjunctivitis.

The BioStar OIA Chlamydia test (BioStar, Inc., Boulder, Colo.) is a novel immunoassay system that uses changes in reflection of light to directly detect chlamydial antigen in clinical specimens. We compared the optical immunoassay (OIA) with culture for detecting Chlamydia trachomatis in ocular specimens from infants with suspected chlamydial conjunctivitis. We initially performed a retrospective evaluation, testing 152 ocular specimens previously collected for culture with the OIA. The sensitivity and specificity were 94.2 and 97%, respectively. A subsequent prospective study evaluating 37 ocular specimens from infants with suspected C. trachomatis conjunctivitis revealed a sensitivity and specificity of 100 and 92.6%, respectively.     

Evaluation of the digene hybrid capture II CT-ID test for detection of Chlamydia trachomatis in endocervical specimens

The performance characteristics of the new signal amplification-based Hybrid Capture (HC) II CT-ID test system (Digene, Silver Spring, Md.) with endocervical specimens were compared to those of tissue culture and PCR (AMPLICOR CT PCR; Roche Molecular Systems, Branchburg, N.J.) for detection of Chlamydia trachomatis in 587 women. HC II CT-ID identified 62 of 65 confirmed C. trachomatis-positive patients (sensitivity of 95.4%) and was negative for 517 of 522 patients who were negative by culture and PCR (specificity of 99.0%). Twelve of the 65 confirmed positive patients were negative by culture but were identified by both HC II CT-ID and PCR (sensitivity of culture was 81.5% [P < 0.01]). In comparison, PCR detected 59 of 65 positive specimens (sensitivity of 90.8%) and had a specificity of 99.6% (520 of 522). These results demonstrate that the Digene HC II CT-ID test is a highly sensitive and specific assay for the detection of C. trachomatis infection in endocervical specimens.     

Comparison of detection procedures for Chlamydia trachomatis, including enzyme immunoassays, in a mouse model of genital infection

Two chlamydial enzyme immunoassays, Chlamydiazyme and IDEIA, were evaluated in a mouse model of chlamydial genital-tract infection. The Chlamydiazyme assay and the IDEIA were assessed on specimens from 10 and 11 mice, respectively. The animals were infected with Chlamydia trachomatis, strain SA2f, and the results obtained by these methods on vaginal specimens taken on 4 or 5 occasions during 41-42 days were compared with those obtained in cell culture and to a less extent by the MicroTrak direct immunofluorescence test. In comparison with culture, the Chlamydiazyme assay had a sensitivity of 62% and a specificity of 92%; IDEIA had a sensitivity of 76% and a specificity of 94%. These assays sometimes did not detect chlamydiae in specimens taken immediately before specimens which proved positive by culture and the immunoassays were less sensitive if swabs were taken after those for culture. The IDEIA also failed to detect chlamydiae in the late stage of the murine infection when chlamydial elementary bodies were seen by immunofluorescence. The implications of the observations for investigations in the human field as well as for further studies in the mouse are discussed.     

Detection of Chlamydia trachomatis in first-void urine collected from men and women attending a venereal clinic

Cervical, urethral and first-void urine (FVU) specimens from 196 men and 245 women attending a venereal outpatient clinic were studied by culture and a commercial enzyme immunoassay (EIA) (CHLAMYDIAZYME). Confirmatory chlamydial testing by a direct fluorescence assay (DFA) (MICROTRAK) was performed on the sediments of the positive EIA samples from culture-negative patients. Chlamydia trachomatis was isolated from 11% of the men and 12% of the women. Of the women, 67% were positive in both sampling sites and 33% in the cervix only. No further cases were found when a female urethral swab was cultured. All the chlamydia-positive urine samples were obtained from women who were positive in the urethra. The denominator used to calculate sensitivities was the combination of patients with culture- and EIA-positive results which could be confirmed by DFA. The sensitivity of our culture method was 85% for men and 77% for women. In men, the sensitivity of EIA was greater on urine than on urethral specimens (77% vs 62%; p less than 0.1). In women, the sensitivity of EIA on urine was significantly poorer than that on cervical specimens (54% vs 85%, p less than 0.001). The specificity of EIA ranged between 94 and 100%. Our study suggests that it may be worth using FVU in a trial for the diagnosis of genital chlamydial infections in symptomatic men, but not in symptomatic women.     

Comparison of manual Amplicor PCR, Cobas Amplicor PCR, and LCx assays for detection of Chlamydia trachomatis infection in women by using urine specimens.

We compared the Roche Amplicor PCR, Roche Cobas Amplicor PCR, and Abbott LCx assays by using urine specimens for the detection of Chlamydia trachomatis infections in a female population. First-catch urine and endocervical swab specimens were collected from a total of 442 patients. Urine specimens were tested by the manual Roche Amplicor PCR, the automatic Roche Cobas Amplicor PCR, and the Abbott LCx assays as instructed by the manufacturers. For the Cobas Amplicor PCR, the internal control protocol was used for every specimen to reveal the presence of polymerase inhibitors. Cell culture of cervical specimens was used as a reference method. Of 442 patients, 50 (11.3%) were confirmed to have chlamydial infection. The diagnostic sensitivity and specificity of cell culture with cervical swab specimens were 88 and 100%, respectively. With urine specimens the sensitivity and specificity for the manual Amplicor PCR assay were 100 and 99.7%, respectively; those for the automatic Cobas Amplicor PCR assay were 94 and 99.2%, respectively; and those for the LCx assay were 94 and 100%, respectively. Thus, all amplification methods with urine specimens proved to be highly sensitive and specific for the detection of C. trachomatis infection in women. No statistically significant differences in the test performances could be demonstrated for specimens from this population. All three amplification techniques with urine specimens proved to be superior to cell culture with cervical swab specimens in diagnosing C. trachomatis infections in women.     

Enzyme immunoassay for the detection of Chlamydia trachomatis antigen in urethral and endocervical swabs.

An enzyme immunoassay technique based on the direct detection of Chlamydia trachomatis antigen in urethral or cervical swabs was used for the rapid diagnosis of chlamydial genital infection. Urethral and cervical samples from 140 patients were tested in parallel by enzyme immunoassay and cell culture using iodine staining. The direct test had a sensitivity of 92.5% and specificity of 97.2% when compared with the cell culture system. The enzyme immunoassay technique provides a rapid and simple method for diagnosing chlamydial genital infection and may be performed on a large number of samples in laboratories which do not have tissue culture facilities or a trained microscopist.