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We evaluated performance of Abbott RealTime CT/NG assay (real-time PCR, Abbott Japan) for detect Chlamydia trachomatis and Neisseria gonorrhoeae by real-time PCR in 88 female patients with cervicitis symptoms seen at gynecological clinics and 100 male patients with urethritis symptoms seen at urological or dermatology clinics in Kitakyushu, Japan. Endocervical swab and first-voided urine (FVU) specimens were then collected from women and FVU specimens from men. Detection rates of C. trachomatis and N. gonorrhoeae by real-time PCR in the 3 types of specimens were compared to those by ProbeTec ET assay (ProbeTec, BD Diagnostic System). The overall positive concordance between real-time PCR and ProbTec were 97.1% (66/68) for C. trachomatis and 100% (33/33) for N. gonorrhoeae, C. trachomatis detection yielded 3 discordant results in endocervical specimens and 1 discordant result in male FVU by real-time PCR and ProbTec. Three of 4 reexamined using Aptime Combo 2 Assay (Fuji Rebio Inc.) were positive for C. trachomatis. Endocervical swab and FVU specimen results for C. trachomatis were discordant in 3 cases in real-time PCR and 4 in ProbeTec. Subjects with 2 or more positive endocervical awab results in female or male FVU specimens were assumed to be "true positive" for C. trachomatis. The sensitivities of real-time PCR for detecting C. trachomatis was 94.4% in endocervical swabs, 77.8% in female FVU and 97.4% in the male FVU. The sensitivities for real-time PCR for detectig N. gonorrhoeae was 100% in all 3 specimen types. Abbott RealTime CT/NG assay was useful for detecting C. trachomatis using endocervical swabs or male FVU specimens and for detecting N. gonorrhoeae using endocervical swabs and all FVU specimens.

Objectives: Serum levels of antibody against Chlamydia trachomatis are used routinely to diagnose chlamydial infection among patients undergoing counseling and testing for sexually transmitted disease at public health centers in Japan, whereas tests for pathogen detection are conducted at most clinics. The objective of the present study was to evaluate the usefulness of a test for detecting C. trachomatis and to compare it with that of screening for serum chlamydia antibodies at a public health center., Methods: Serum chlamydia antibody titers were estimated using an ELISA, while C. trachomatis was detected in the urine using the nucleic acid amplification assay that included strand displacement amplification (SDA). We compared the results of 120 patients (64 men and 56 women), because the sensitivity of the method using urine samples from women has been reported to be as high as that using cervical samples. The serum antibody test results were considered to be positive when either one or both of the IgA and IgG antibodies were positive., Results: The overall prevalence of positive results for the antibody test was 24.2% (14.1% for men and 35.7% for women), and that for the SDA method was 7.5% (3.1% for men and 12.5% for women). The concordance rate of the 2 tests was 81.7%, with a kappa statistic of 0.35 (95% CI, 0.10-0.59). Of the 9 subjects who were positive for C. trachomatis using the SDA assay, 1 had serum chlamydia IgA antibodies, 6 had IgG antibodies, 1 had both the antibodies, and 1 had no antibodies. On the other hand, of the 111 subjects who were negative for C. trachomatis, only 8 had chlamydia IgA antibodies, 5 had IgG antibodies, and 8 had both the antibodies. These results indicated there was no association between serum IgA antibodies and the presence of C. trachomatis. The presence of serum chlamydia antibodies was significantly higher in subjects with a past history of genital chlamydial infection than in those without a past history of infection (P<0.01). However, there was no difference in the results of the SDA assay between subjects with or without a previous infection., Conclusion: Thus, although serum chlamydia antibodies are generally measured at public health centers, tests for detecting C. trachomatis should also be used.

The clinical efficacy and safety of levofloxacin (LVFX) 500mg qd were evaluated in female patients with cervicitis with Chlamydia trachomatis and intrauterine infections. LVFX was administered orally at 500 mg qd for 7 days. Bacteriological efficacy was 94.4% (17/18) and clinical efficacy was 100% (16/16) at 14 to 21 days after the end of treatment in cervicitis. On the other hand, bacteriological efficacy and clinical efficacy at the end of treatment in intrauterine infections were 68.8% (11/16) and 94.7% (18/19), respectively. For safety, adverse drug reactions occurred in 9 of 43 patients (20.9%), i.e., increased y-GTP in 2 patients, glucose urine present in 2, and each of all other adverse reactions occurred in 1. All adverse drug reactions observed were either mild or moderate. Results suggested that LVFX 500 mg qd was effective and safe in the treatment of cervicitis with Chlamydia trachomatis and intrauterine infections.

We evaluated performance of Abbott RealTime CT/NG assay (real-time PCR, Abbott Japan) for detect Chlamydia trachomatis and Neisseria gonorrhoeae by real-time PCR in 88 female patients with cervicitis symptoms seen at gynecological clinics and 100 male patients with urethritis symptoms seen at urological or dermatology clinics in Kitakyushu, Japan. Endocervical swab and first-voided urine (FVU) specimens were then collected from women and FVU specimens from men. Detection rates of C. trachomatis and N. gonorrhoeae by real-time PCR in the 3 types of specimens were compared to those by ProbeTec ET assay (ProbeTec, BD Diagnostic System). The overall positive concordance between real-time PCR and ProbTec were 97.1% (66/68) for C. trachomatis and 100% (33/33) for N. gonorrhoeae, C. trachomatis detection yielded 3 discordant results in endocervical specimens and 1 discordant result in male FVU by real-time PCR and ProbTec. Three of 4 reexamined using Aptime Combo 2 Assay (Fuji Rebio Inc.) were positive for C. trachomatis. Endocervical swab and FVU specimen results for C. trachomatis were discordant in 3 cases in real-time PCR and 4 in ProbeTec. Subjects with 2 or more positive endocervical awab results in female or male FVU specimens were assumed to be "true positive" for C. trachomatis. The sensitivities of real-time PCR for detecting C. trachomatis was 94.4% in endocervical swabs, 77.8% in female FVU and 97.4% in the male FVU. The sensitivities for real-time PCR for detecting N. gonorrhoeae was 100% in all 3 specimentypes. Abbott RealTime CT/NG assay was useful for detecting C. trachomatis using endocervical swabs or male FVU specimens and for detecting N. gonorrhoeae using endocervical swabs and all FVU specimens.

Spondyloarthritis (SpA) includes reactive arthritis (ReA) and enteropathic arthritis (EA), which are clinically important but often misdiagnosed. ReA, sterile inflammatory arthritis, arises after certain genitourinary or gastrointestinal infections. Chlamydia are the most common pathogens causing ReA; ReA due to Chlamydia infection is called Chlamydia-associated arthritis (Chl-AA). Recently, Chlamydia trachomatis was detected in the synovial tissue from patients with ReA by electronmicroscopy. In addition, mRNA as well as DNA has been detected in the synovial tissue, suggesting that Chlamydia are viable in inflamed joints. Thus, the notion that ReA is a sterile inflammation should be reconsidered. Chl-AA patients, especially women, often show no symptoms and signs of genitourinary infection. Thus, Chl-AA should be suspected in patients with inflammatory arthritides that is difficult to diagnose. EA is accompanied by inflammatory bowel diseases (IBD). In Japan, over 130,000 individuals have IBD; IBD is diagnosed in 6,500 individuals every year. Around 10% IBD patients develop arthritis, suggesting that 13,000 patients develop arthritis every year. SpA includes peripheral and axial arthritis; axial arthritis includes spondylitis and sacroiliac arthritis. Sacroiliac joint tests need to be performed to diagnose sacroiliac arthritis. Rheumatologists should be aware of the pathogenesis of Chl-AA and EA and diagnose and treat these diseases appropriately.

Urine samples collected from 422 males and 53 females visiting a clinic in Kawasaki City who were suspected to have sexually transmitted infection were tested for the detection of Chlamydia trachomatis and Neisseria gonorrhoeae by BD ProbeTecET (SDA method). The detection rates of C. trachomatis by the SDA method and polymerase chain reaction (PCR) method (control) were as high as 98.1% for C. trachomatis, and as high as 99.4% for N. gonorrhoeae, and the concordance rate of detection of both bacterial species was high. The detection sensitivity and specificity of the SDA method were 90.6 and 99.3%, respectively for C. trachomatis and 98.7% and 100% for N. gonorrhoeae, when PCR was used as the standard method. There were no differences in these results between males and females. The number of patients showing a discrepancy of the results obtained between the SDA method and the PCR method was 9 for C. trachomatis and 1 for N. gonorrhoeae, but the results of redetermination by the SDA method tended to coincide with those of the PCR method. Urine samples tested by the SDA method were positive for N. gonorrhoeae even in patients in whom the culture of secretions from the male urethra was negative for N. gonorrhoeae. Based on these results, the BD ProbeTecET (SDA method) was confirmed to have the equivalent capability to the PCR method for the detection of C. trachomatis and N. gonorrhoeae in urine samples.

The performance of a real-time DNA amplification assay, BD ProbeTec ET System (BDPT, BD Diagnostic Systems), to detect Chlamydia trachomatis and Neisseria gonorrhoeae on endocervical and oropharyngeal samples was evaluated. After obtaining informed consent, 364 endocervical, 363 urine and 247 oropharyngeal specimens were collected from 307 cases. The overall agreement rate of the BDPT and Amplicor (AMP, Roche) assays for the detection of C. trachomatis and N. gonorrhoeae in endocervical samples was 99.2% (361/364) for C. trachomatis and 99.5% (362/364) for N. gonorrhoeae. Assay of oropharyngeal swabs by the BDPT yielded 21 C. trachomatis positives, and 19 of them were C. trachomatis negative by the DNA probe assay (Gen-Probe PACE). The AMP assay showed that 16/19 (84.2%) of the BDPT +/DNA probe - samples were positive. The BDPT also yielded 21 N. gonorrhoeae positives, 15 of which were negative with the DNA probe. Additional testing showed that all 15 BDPT +/DNA probe - samples were positive by the established nested PCR method. Our data suggest that the performance of the BDPT is comparable to that of AMP for detection of C. trachomatis and N. gonorrhoeae in endocervical swab samples and that it may be a useful method for detecting of C. trachomatis and N. gonorrhoeae in oropharyngeal samples clinically.

The data of sexually transmitted urethritis in males have been collected at 24 institutes in Kyoto Prefecture since October, 2002. The data collected from January to December in 2004 are summarized herein. A total of 1,275 patients were diagnosed with urethritis during this period. Microbiological examinations isolated Neisseria gonorrhoeae alone in 368 (29%), Chlamydia tracomatis alone in 336 (26%), both in 85 (7%), and others in 453 (36%). Male patients under 20 years old tended to have Chlamydial urethritis, alone or combined with gonococcal infection, and had a predominant infectious source, a non-commercial-sexual-worker female partner, suggesting a profound problem in sexual life of adolescents. The urologist preferred to use quinolones as the first therapeutic modality against male urethritis. However, drug resistance of N. gonorrhoeae, especially against quinolones, has rapidly progressed, which was also observed by a sensitivity examination test. Antibiotics should be used adequately against male urethrits according to the recent guidelines.

The adherence to and invasion of A549 cells by serotype VIII GBS (group B streptococci) were significantly greater than those of serotype III GBS for both the invasive strain and vaginal isolates. Serotype III strains of GBS stimulated significantly greater tumor necrosis factor alpha (TNF-alpha), interleukin-10 (IL-10), and interleukin-12 (IL-12) production than did serotype VIII strains. Interleukin-4 (IL-4) and interleukin-8 (IL-8) production in response to serotype VIII was significantly higher than in response to serotype III. On sexual transmitted diseases, orogenital contact has been common in both commercial sex workers (CSWs) and non-CSWs in Japanese women. Therefore, chlamydial infections in pharynx have been increasing in recent days. However, the treatment against Chlamydia trachomatis in the cases of pharyngeal infections should be longer than in the cases of uterine cervical infections.

The isothermal and chimeric primer-initiated amplification of nucleic acids (ICAN) is a new isothermal DNA amplification method composed of exo Bca DNA polymerase, RNaseH and DNA-RNA chimeric primers. We developed the simultaneous detection system for Chlamydia trachomatis/Neisseria gonorrhoeae DNA, combined with luminescence detection by a probe hybridization. In the performance tests, this system was able to detect 10 to 100 copies of C. trachomatis/N. gonorrhoeae DNA for only 3.5 hours, and was highly specific to C. trachomatis/N. gonorrhoeae without any cross-reaction to C. pneumoniae, N. lactamica, N. sicca or N. meningitidis. When we tested 60 clinical samples of urine and cervical swabs, the interpretive results were completely consistent with those obtained by Roche PCR system. Of 13 positive samples by the ICAN and PCR systems for C. trachomatis, four were negative by EIA method(IDEIA Chlamydia). These results indicate that the ICAN system is an efficient and sensitive system to simultaneously detect C. trachomatis/N. gonorrhoeae DNA.

Purpose: We reviewed the clinical data of male urethritis diagnosed as sexually transmitted disease with the aim of revealing its clinical features., Patients and Methods: In a total number of 414 males diagnosed as having acute urethritis transmitted by sexual activities between January and December in 2000, clinical data were collected and analyzed. In addition, questionnaire concerning the awareness of pharyngeal infection was obtained., Results: Their ages ranged from 16 to 60 years with a mean of 31 years. The source of infection was a commercial sexual worker (CSW) in 288 (69.6%) out of 414 cases. In 241 cases in whom an infection route was determined, 199 cases (82.6%) were considered to have an infection through oral sex. Microbiological examinations isolated Neisseria gonorrhea alone in 206 (49.8%), Chlamydia tracomatis alone in 47 (11.3%) and both in 46 (11.1%) cases. In our study, 57.5% and 20.8% of Neisseria gonorrhea strains were resistant to penicillin and new quinolones, respectively. Based on questionnaire, 174 (57.8%) out of 301 cases were not aware of possible infection from the pharynx through oral sex., Conclusion: The remarkable prevalence of gonococcal urethritis might be in part due to the lacking of awareness of infection through oral sex and sexual activities without a condom. Instruction on the prevention as well as the actual situation of urethritis is needed for general population with a broad age spectrum. The appropriate use of antimicrobial agents with sufficient susceptibility and less inductivity of drug-resistance is also of clinical importance.

Between 1992 and 1998, serotyping of 82 Chlamydia trachomatis cervical isolates were examined by micro-IF method in Nagano Prefecture. Of these, 17 isolates (20.7%) were serovar E, 15 (18.3%) were serovar F, 14 (17.1%) were serovar D and 10 (12.2%) were serovar G, and the isolates typed these serovars were found to be 68.3% whole. Furthermore, serovars B (7.3%) and K (9.8%) were comparatively frequently found. From 1992 to 1994, the number of isolates of B-complex, intermediate and C-complex were 18, 17 and 6, respectively. On the other hand, from 1995 to 1998, the number of the isolates were 19, 8 and 14, respectively. The distribution of serovars of C. trachomatis tended to fluctuate from serovars D and E to F and G as the patients grew older. Moreover, serovar E isolates were only detected from patients less than 40 years old. Although most of the serovar I isolates were detected from the patients with vaginal discharge, the other isolates did not clearly indicate the relationship of serovars and clinical symptoms.

Chlamydia trachomatis detection by both swab CHLAMYDIAZYME and urine sediment AMPLICOR were performed on 61 males and 54 females who were suspected to have C. trachomatis genital infection (CTGI). Positive agreement between swab CHLAMYDIAZYME and urine sediment AMPLICOR was good, 96.9% in males and 96.6% in females. Negative agreement was 100% in males and 84.0% in females. "Swab CHLAMYDIAZYME positive-Urine sediment AMPLICOR negative specimen" were not obtained. "Swab CHLAMYDIAZYME negative-urine sediment AMPLICOR positive" resulted in four females who were finally diagnosed to have CTGI by repeated specimens on CHLAMYDIAZYME. Positive result by AMPLICOR was obtained on un-spun urine specimen that was in agreement with the laboratory result that the sensitivity of AMPLICOR reaches one thousand fold of CHLAMYDIAZYME. Detection on every separately collected urine specimen of one urination showed AMPLICOR positive all through the last cup specimen in both male and female. Those results seem to certify the reliability of C. trachomatis detection by urine sediment AMPLICOR. False negative by interference substance in urine and false positive by contamination were not experienced. On asymptomatic young male and female, positive rates of urine sediment AMPLICOR were 7.5% in male and 12.1% in female which are far more than the positive rate obtained by CHLAMYDIAZYME. Non invasive C. trachomatis detection method applicable to asymptomatic female is needed for control of CTGI. C. trachomatis detection could be facilitated by non invasive urine sediment AMPLICOR.

The initial site of infection with female chlamydia trachomatis (CT) genital infection is known to be the cervix. CT can often be detected in vaginal discharge, urethral swabs and urinary sediment. In order to determine the distribution of CT around the cervix, CT detection by Chlamydiazyme was performed on specimens other than cervical swabs, obtained from females whose cervical swabs were positive for CT. Positive rates and mean reaction values for Chlamydiazyme were 97.4% and 0.762 +/- 0.570 for cervical discharge, 75.4% and 0.503 +/- 0.536 for vaginal wall swab, 72.1% and 0.489 +/- 0.516 for vaginal swab, 62.7% and 0.696 +/- 0.760 for urethral swab and 43.8% and 0.228 +/- 0.328 for urinary sediment. Both the positive rate and the mean reactive value were revealed to be in the order: vaginal wall = vestbule swab < vaginal discharge < cervical swab. The results are consistent with the assumption that CT was contained in discharge in the infected area in the cervix and spread to other areas. Both a high positive rate and high reactive value for vaginal discharge sustain the assumption that CT rectal infection could mainly be caused by direct contamination by cervical discharge, not by rectal intercourse.

Specimens from 15 young patients presenting with acute epididymitis were tested for the presence of Chlamydia trachomatis by an enzyme immunoassay (EIA), polymerase chain reaction (PCR), and for other bacteria by standard laboratory techniques. C. trachomatis urethral infection was detected in 3 patients by an EIA test of the urethral swabs (20%) and in 13 patients by the PCR (87%). This difference in detection rate was statistically significant (p < 0.005). Thirteen specimens were positive by the PCR, but only three of them were positive by the EIA method. These findings indicate that the PCR assay is a highly sensitive assay for the detection of C. trachomatis in male urine specimens and provides a noninvasive technique for routine screening of chlamydia infection in the patient with acute epididymitis.

We analysed the clinical background of female who had no underlying disease but consulted out urology outpatient clinic with a chief complaint of symptoms of bladder irritation. Overally, 50.1% (102/200 cases) of the cases had urethral syndrome, without pyria or bacteriuria. Plotting of the age distribution of the urethral syndrome cases showed a bell-shaped curve, with the peak at the 40-49 year-old age bracket. The age distribution for the cystitis cases showed two peaks: at 20-29 years of age and 60-69 years of age. These results indicate that the cases of urethral syndrome involve multiple factors with increasing age. Examination of the external genitalia revealed the urethral smear specimens to be positive for leukocytes in 48.9% (88/180 cases) of the total urethral syndrome cases and in 72.7% (16/22 cases) of the urethral syndrome cases positive for urethral bacteria. Accordingly, it was surmised that there is considered to be a precursor of cystitis-as a cause of urethral syndrome cases. In addition, the rates of detection of chlamydia trachomatis from the vagina and urethra of the urethral syndrome cases in the reproductive age range of (2/34 cases) for the urethra. However, these rates were about the same as those determined for healthy volunteers. Nevertheless, it was surmised that urethritis or vaginitis caused by C. trachomatis was one of the causes of the symptoms of bladder irritation in some of the cases. Accordingly, it was concluded that infection due to C. trachomatis cannot be ignored as a possible cause of urethral syndrome.

Mycoplasma genitalium causes urethritis in non-human primates, but studies on its pathogenicity in man have been hampered by the difficulty in isolating this oragnism in culture. We have used a specific polymerase chain reaction to examine the role of M. genitalium in non-gonococcal urethritis (NGU). Oligonucleotide primers were used to amplify a 281 bp of 140-KDa adhesin gene of M. genitalium. A characteristic PCR product was amplified, when M. genitalium DNA was template for the PCR. No amplified product was detected in Mycoplasma pneumoniae DNA, Mycoplasma hominis DNA or other bacterial DNAs. M. genitalium DNA was detected in urethral swabs from 17 (14.9%) of 114 men with NGU. Three (9.1%) of the 33 men with Chlamydia-positive NGU and 14 (17.3%) of the 81 with Chlamydia-negative NGU were positive for M. genitalium DNA, but 29 men without urethritis were negative. The prevalence of M. genitalium in NGU and in Chlamydia-negative NGU was significantly higher than that in the normal control. These findings suggest that M. genitalium would be a cause of NGU.

In an attempt to know the applicability of non-cultural detection technique for N. gonorrhoeae and C. trachomatis on pharyngeal and rectal swabs, the swabs taken from N. gonorrhoeae- and C. trachomatis-free subjects were tested by Gonozyme, Chlamydiazyme, Ideia Chlamydia, Gen-Probe and Gen Probe Pace 2. As N. gonorrhoeae- and C. trachomatis-free subjects, the castrated males older than age 70, with known prostate cancer treated at least for 6 months with anti-androgens who were seronegative for C. trachomatis and had a history of Ofloxacin treatment in the last 3 months and whose pharyngeal and rectal swabs were negative for N. gonorrhoeae, were subjected for study. The false negative rates were for N. gonorrhoeae from the pharynx by Gonozyme 57.7%, by Gen-Probe 0%, by Gen-Probe Pace 2 0%, from rectum by Gonozyme 0%, by Gen-Probe 0% by Gen-Probe Pace 2 0%, for C. trachomatis from pharynx by Chlamydiazyme 93.8%, by Ideia Chlamydia 8.3%, by Gen-Probe 0%, by Gen-Probe Pace 2 0%, from rectum by Chlamydiazyme 94.4%, by Ideia Chlamydia 70.9%, by Gen-Probe 20.0% and by Gen-Probe Pace 2 0%. To investigate the incidence of pharyngeal and rectal infection by N. gonorrhoeae among the patients with gonococcal genital infection, as well as pharyngeal and rectal infection by C. trachomatis among the patients with chlamydial genital infection, pharyngeal and recal swabs obtained from the patients with genital infection by N. gonorrhoeae or C. trachomatis were tested by Gen-Probe Pace 2.(ABSTRACT TRUNCATED AT 250 WORDS)

Recently, two new kits, HITAZYME (Hitachi Chemical Co., Ltd.) and SERO IPALISA (Savyon Diagnostics, Ltd.), for the assay of anti-C. trachomatis antibodies by the enzyme immunoassay (EIA) method have been developed and put into clinical application. In the study reported here, the authors investigated the clinical usefulness of these assay kits, together with the IPAzyme and micro-IF test, in the diagnosis of cases of urogenital tract C. trachomatis infections. 1. The positive rates for IgA antibodies, which are considered to be an indicator of active infection, obtained with the HITAZYME and SERO IPALISA kits in the 82 antigen-positive cases were significantly (p < 0.005) higher than the rates obtained with the IPAzyme and micro-IF test. These results showed the usefulness of the HITAZYME and SERO IPALISA kits for detecting C. trachomatis infections. 2. A comparison was made of the assay results obtained with the HITAZYME and SERO IPALISA kits, and it was found that there was a large number of cases (142) that tested negative for IgA antibodies with the HITAZYME but positive with the SERO IPALISA kit. We carried out a confirmatory test on the specimens of cases for which the results obtained with the HITAZYME and SERO IPALISA kits were not in agreement. This test employed the Western blotting method using COMC (the antigen extracted from EB of C. trachomatis strain L2 and used in the HITAZYME kit) and whole EB of C. trachomatis strain L2 (the antigen used in the SERO IPALISA kit). The results showed a significantly higher degree of agreement between the HITAZYME kit data and the Western blotting data than between the SERO IPALISA kit data and the Western blotting data. 3. In addition, with the objective of investigating the existence of cross reactivity with anti-C. pneumoniae antibodies, we performed Western blotting using as the antigen crude whole EB of C. pneumoniae strain TW-183. The results showed that anti-C. pneumoniae antibodies were detected in 25 of 35 (71.4%) cases that were negative with C. trachomatis antigen and the HITAZYME kit and positive with the SERO IPALISA kit. These findings indicate a strong possibility that these cases positive with the SERO IPALISA kit are due to a cross reaction with anti-C. pneumoniae antibodies.(ABSTRACT TRUNCATED AT 400 WORDS)

Untreated infection of female partners by men with chlamydial epididymitis may have serious effects on the partners' fertility. To assess the need for detailed microbiological investigation, 32 patients, 35 years old or younger, with epididymitis and their sexual partners were examined. The patients underwent thorough evaluations, including chlamydia isolation, microscopy of urethral swab, bacterial culture, and chlamydia serologic testing. An infective cause was identified in 56% of the patients. The most common microorganism was Chlamydia trachomatis. This microorganism was identified from urethral swabs in 11 patients (34%). A total of 18 sexual partners were traced and investigated for chlamydia antigen by cervical swab, urinary bacterial culture, and chlamydia serologic testing. Of the 18 female sexual partners screened, 9 were partners of patients with chlamydial epididymitis; 78% of these women had the same infection. Young men with epididymitis, as well as their partners, should undergo full microbiological evaluation including Chlamydia trachomatis for adequate treatment of this infection.

The intensity of reaction in the IDEIA CHLAMYDIA test kit to lipopolysaccharide of seven strains of Chlamydia pneumoniae was examined by using highly purified elementary bodies (EBs). The strains were divided into two groups according to whether the EBs were pear-shape or round. The group with the pear-shaped EBs consisted of TW-183, AR-39 and AR-388 strains; the group with the round EBs consisted of IOL-207, Kajaani-6, YK-41 and KKpn-1 strains. The number of EBs at the cutoff level of the test kit were determined as follows: TW-183, 7.0 x 10(3); AR-388, 8.4 x 10(3); AR-39, 2.4 x 10(4); IOL-207, 6.0 x 10(3); Kajaani-6, 1.2 x 10(4); KKpn-1 2.8 x 10(4); and YK-41, 4.0 x 10(4) per assay. The number of EBs of C. trachomatis L2/434/Bu and C. psittaci Cal 10 were 1.0 x 10(3) and 2.7 x 10(4) per assay, respectively. The results demonstrated that the EBs of C. pneumoniae were detected with the test kit at different reaction intensities ranging from 6.0 x 10(3) to 4.0 x 10(4) per assay and that there was no correlation between EB number and morphology.

We have reported a method for detection of Chlamydia trachomatis by polymerase chain reaction (PCR) with two oligonucleotides based on sequences within the major outer membrane protein gene from C. trachomatis serovar L2. In the previous report, in addition to treatment of the mixture of first-voided urine (FVU) sediment and 1 ml of urine with proteinase K. DNA purification by phenol extraction was necessary for preparation of template DNA for PCR. In this study, FVU sediment was suspended in 1 ml of Chlamydiazyme dilution buffer and a part of the suspension was treated with proteinase K for DNA extraction. The DNA extraction solution could be used as template for PCR without purification of DNA by phenol extraction. One hundred FVU specimens obtained from male urethritis patients were examined with the two methods (PCR and IDEIA) for detection of C. trachomatis. In 33 of 100 specimens, the DNA fragments of C. trachomatis was amplified by the PCR and in 32 of 100, the chlamydial antigen was detected by IDEIA. The positive and negative coincidence rate of the PCR to IDEIA were 93.8% (30.32) and 95.6% (65/68) respectively, resulting in a high overall coincidence rate at 95%. Thus, the improved method with PCR using FVU as a specimen is proved to be a useful, non-invasive diagnostic tool for diagnosis of chlamydial urethritis.

Neisseria gonorrhoeae was detected from first-voided urine sediments of male patients with urethritis by polymerase chain reaction (PCR). Urine and urinary sediment were treated with proteinase K, and DNA was further purified by phenol extraction. Two oligonucleotides based on sequences within a ribosomal RNA gene from N. gonorrhoeae were used as primers for the PCR. A DNA fragment of 206 bp specific for N. gonorrhoeae was amplified by PCR and detected by agarose gel electrophoresis. In 19 specimens of urine sediments collected from 21 patients in whom N. gonorrhoeae was isolated from urethral swab by culture, 206 bp DNA fragment was amplified by PCR. In all specimens of urine sediments from 24 patients in whom cultures for N. gonorrhoeae were negative, no DNA was amplified by the PCR. The overall coincidence rate between the PCR for detecting N. gonorrhoeae in first-voided urine sediments and culture in urethral swab was 95.6% (43/45). PCR procedure for detection of pathogens from first-voided urine sediments would be noninvasive and would be applied for the diagnosis of gonococcal urethritis and chlamydial urethritis.

Chlamydia trachomatis was detected from first-voided urine sediments of 97 male patients with urethritis by polymerase chain reaction (PCR). Since urine and urinary sediments only treated with proteinase K inhibited DNA amplification by PCR, DNA was further purified by phenol extraction and concentrated. Two oligonucleotides based on sequences within the major outer membrane gene from C. trachomatis serovar L2 were used as primers. A DNA fragment of 242 bp specific for C. trachomatis was amplified by PCR and detected by agarose gel electrophoresis. The DNA fragment was amplified by PCR in all specimens of urine sediments from 50 patients with Chlamydiazyme-positive urethral swab. In 38 specimens of urine sediments from 47 patients with Chlamydiazyme-negative urethral swab, PCR was negative. The overall coincidence rate between the PCR for detecting C. trachomatis in first-voided urine sediments and Clamydiazyme in urethral swab was 90.7% (88/97). Detection of C. trachomatis from first-voided urine sediments by PCR was considered to be noninvasive and useful for the diagnosis of male urethritis due to C. trachomatis.

A method using polymerase chain reaction (PCR) was compared to an enzyme immunoassay (Chlamydiazyme) for detection of Chlamydia trachomatis by testing a reference strain and clinical specimens. Two oligonucleotides based on sequences within the major outer membrane protein gene from C. trachomatis serovar L2 were used as primers for the PCR. A DNA fragment of 242 bp specific for C. trachomatis was amplified by the PCR, when DNA of greater than or equal to 10(2) C. trachomatis was used as template for the PCR. A chlamydial antigen was detected by Chlamydiazyme, when greater than or equal to 2.6 x 10(3) C. trachomatis were applied for the enzyme immunoassay. The PCR method was 26 times more sensitive than Chlamydiazyme in detection of C. trachomatis. The PCR method and Chlamydiazyme were carried out to examine 74 urethral swabs obtained from male patients with urethritis for detection of C. trachomatis. In 45 of 74 specimens, the DNA fragment of C. trachomatis was amplified by the PCR, and in 41 of 74, the chlamydial antigen was detected by Chlamydiazyme. The detection rate of the PCR method (60.8%) was higher than that of Chlamydiazyme (55.4%). The positive coincidence rate of the PCR method to Chlamydiazyme was 100% (41/41) and negative coincidence rate was 87.9% (29/33). The overall coincidence rate between the two methods was high (94.6%). Thus, the PCR method was more sensitive than Chlamydiazyme for detection of C. trachomatis and specific for diagnosis of chlamydial urethritis.

The author devised a method which permits simultaneous isolation and identification of Chlamydia using a slide chamber with 8 wells. Contaminating bacteria were eliminated by filtration with the membrane filter. The procedure allowed isolation of Chlamydia pneumoniae and Chlamydia trachomatis from 11 (10.1%) out of 109 and 10 (9.2%) out of 109 otolaryngologic clinical specimens, respectively. The use of HL cells together with Hela-229 cells had been considered essential to isolate Chlamydia from specimens in this field. The conditions for isolating Chlamydia pneumoniae were similar to those for Chlamydia trachomatis. The slide chamber procedure with membrane filter treated specimens was suitable for simultaneous isolation and culture of various species of Chlamydia and allowed easy differentiation of Chlamydia pneumoniae from Chlamydia trachomatis with identification by the indirect fluorescent-antibody technique. Routine examination of clinical specimens using this procedure should be required in the future in order to better understand chlamydial infections.

A polymerase chain reaction (PCR) with Chlamydia trachomatis-specific primers was applied for detection of C. trachomatis from urethral swab in male urethritis. The results were compared with those of culture method for detection of C. trachomatis. Of 18 clinical specimens tested in this study, inclusion bodies of C. trachomatis were detected in 11 specimens by the culture method. For PCR, sample DNA was prepared from transport medium in which urethral smear was suspended and two oligonucleotides based on sequences within the major outer membrane protein gene from C. trachomatis serovar L2 were used as extension primers. In 12 of the 18 specimens, 242bp DNA fragment was amplified by PCR and demonstrated to be the DNA fragment of C. trachomatis by Southern blot hybridization. No DNA of 242bp was amplified by PCR from five specimens in which any inclusion bodies of C. trachomatis were observed or from a specimen in which one inclusion body per cover slip was detected by culture method. C. trachomatis DNA of 242bp was amplified from all specimens in which 14 and more inclusion bodies per cover slip were detected by culture method. In two specimens concluded s negative by culture method, amplified C. trachomatis DNA were detected by PCR. Thus, the PCR would be a more simple and sensitive method for detection of C. trachomatis, compared with the culture method.

Three chlamydial strains isolated from patients of otitis media with effusion were studied by comparing reactivity to monoclonal antibody (MAb) and polyclonal antibody (PAb) produced against one clinical isolate (named Mk), which was first isolated by Dr Mukai (Mukai Microbiological Research Laboratory, Yamato-shi, Kanagawa prefecture). Commercially supplied antibody (Microtrak (Syva), Culture-set (Ortho diagnostic system)) was also used. To isolate the Chlamydia spp, the yolk sacs of eggs were immediately inoculated with sample effusions (0.2 to 0.4 ml per sac) as soon as the samples were received. The eggs were observed every day for a period of 12 days thereafter for signs of life or death. One to two blind passages were first done in the eggs and then in HeLa 229 cells. The reactivity was examined by both micro-IF tests, among various strains of Chlamydia (C. trachomatis: L2. C. pneumoniae, C. psittaci: Budgerigar, Izawa, Meningopneumonitis (MP)) and by immunoblot analysis. Chlamydia spp were isolated in two of the twenty-nine sample effusions (6.9%). These isolates were then tested for reactivity to MAb and PAb. It was found that MAb reacted with MP and Mk, but not with Budgerigar, Izawa and C. pneumoniae. The antibody of Culture-set reacted with C. trachomatis C. pneumoniae and C. psittaci. No reactivity was observed in Mk by MicroTrak. Immunoblot analysis revealed that MAb reacted with about 95 KDa protein of Mk, the two clinically isolated Chlamydia spp and MP. By using PAb from rabbits, similar blotting patterns were observed in Mk, the clinical isolates and MP.(ABSTRACT TRUNCATED AT 250 WORDS)

A polymerase chain reaction (PCR) procedure was developed for detection of Chlamydia trachomatis. Two oligonucleotide primers based on sequences within the major outer membrane protein gene from C. trachomatis serovar L2 were used. A single DNA fragment was amplified, when C. trachomatis DNA was template for the PCR. No amplified product was detected in Chlamydia psittaci DNA, Chlamydia pneumoniae DNA or other bacterial DNAs. The amplified DNA fragment was detected, when DNA of greater than or equal to 10(2) C. trachomatis per reaction was used as template for the PCR. Thus, the PCR was shown to be specific for C. trachomatis and more sensitive than the enzyme immunoassays for detection of chlamydial antigen and the chlamydial rRNA:DNA probe hybridization method.

The infections with C. trachomatis and N. gonorrhoeae have recently been determined routinely with the commercial kits for detecting the antigens of these organisms in both the clinical and mass examination laboratories. In mass examination, a change during transporting the specimens must be avoided. Hence, a study on whether the antigens were changed with atmospheric temperature was carried out. As for C. trachomatis, no change in antigen was found on the enzyme immunoassay (EIA) absorbance of the antigen quantity corresponding to the cell of 4.8 x 10(4), 2.4 x 10(4) and 6.0 x 10(3) (IFU/ml) until 5 days at 4, 25 and 37 degrees C respectively after sampling. The coefficient variation was found out to be ca. 10%. In a test on antigen stability of N. gonorrhoeae, similar results to that of C. trachomatis were obtained on the following antigen quantity: 2.4 x 10(3), 3.3 x 10(2) and 9.0 x 10 (CFU/ml) under the same conditions as in the above. The coefficient variation was found out to be 10% or less. To investigate whether there is any difference on the stability of C. trachomatis antigen between mailing and hand carrying, a total of 133 specimens collected from many clinics were subjected to the detection of antigen with the commercial EIA kits. The results of comparison on both transport methods were as follows: agreement rate 96.2%, positive rate 100% and negative rate 95.2%. The highly correlation between the both transportation was confirmed in the detection of antigen; that is, Y = 1.03X + 0.03, r = 0.936.(ABSTRACT TRUNCATED AT 250 WORDS)

To study the incidence of Chlamydia trachomatis (C.T.) infection among women, we obtained swab (endocervical columnar cells) from the uterine cervix of 1,041 women attending the Department of Obstetrics and Gynecology of Gunma University Hospital. The results were as follows. 1) Overall, C.T. was isolated from 60 of the 1,041 women (5.8%). With reference to the diagnoses, the organism was isolated from 38 of 183 patients with cervicitis (20.8%) and from 22 of 858 asymptomatic womens (2.6%) (p less than 0.001). 2) C.T. antigen was detected at a higher frequency (15.0%) in the under 30 years age group than in the over 30 year age group (2.3%) (p less than 0.001). 3) The unstained inclusion C.T. bodies were confirmed more easily by phase contrast microscopy. 4) Inclusion bodies were specified as either reticulate or elementary bodies by the acridine orange staining method.

To study the serovar distribution of C. trachomatis in Japan, a total of 85 genital C. trachomatis isolates from male and female patients attending the clinics were examined by the microimmunofluorescence test using immune sera of the isolates produced in mice. Of these isolates, 34 (40.0%) were typed D or E, and 19 (22.4%) were typed G or F. The serovars of the remaining 32 isolates were B, H, I, J, and K, and the proportions of these serovars were from 8.2 to 3.5%. Thus, two thirds of C. trachomatis isolates in this country were found to fall into only four serovars, namely, D, E, G, and F, and, therefore, the epidemiology of C. trachomatis infection in Japan seems to be similar to that of other countries in North America and Europe. The relative distribution of serovars of C. trachomatis isolates from male patients and female patients somewhat differed. Serovars D, E, and G, F were isolated in the same ratio from male patients, while the isolation ratio of the former serovars was three times or more higher than the latter serovars in female patients. No isolate typed serovar K was found in male patients, while 15% of isolates from female patients were typed this serovar.

We report a case of recurrent tonsillitis and otitis media with effusion (OME) from which Chlamydia trachomatis was isolated. Chlamydia pneumoniae, a newly recognized species of Chlamydia, was also recovered from the tonsillar and bronchial swabs. A 8-year-old girl was seen on February 23, 1988, because of a running nose, a productive cough and bilateral hearing difficulty. She had a history of recurrent tonsillitis. The diagnosis was acute sinusitis with tubal obstruction, then cefixime was prescribed. Her symptoms were once resolved, for the time being but she came back to the hospital a week later with a bilateral ear-ache. The tympanic membranes were injected and characteristically retracted. Her left ear showed type B tympanogram (effusion). Tympanocentesis was performed to remove middle-ear effusion, from which C. trachomatis but no ordinary bacterium was isolated. Therefore rokitamycin 300 mg/day was administered for a week. Her condition improved, however, a rhinorrhea, a plugged ear sensation and a hacking cough returned in a month. She was admitted to the hospital on May 10, for tympanostomy and grommet insertion, but from the day before admission, she had a sore throat with fever (39.2 degrees C). The surgery was withheld until May 26. When adenotonsillectomy and grommets insertion were undertaken, C. trachomatis had disappeared from the middle-ear effusion, but C. pneumoniae was recovered from both tonsillar and bronchial swabs. Readministration of rokitamycin was performed and to date (June, 1990) she remains well.

Sexual behaviour in 141 female adolescents, from 15 to 20 (average 17.4) years old, detained in a remedial institution was investigated from an epidemiological viewpoint, with special reference to Chlamydia trachomatis (CT) infection, from April, 1985 to March, 1986. All had experienced sexual intercourse. Nineteen subjects (13.5%) were found to be CT-antigen positive by either cell cultures (16 cases) or immunofluorescence tests (9 cases). The immunotypes of CT belonged to types D-K. CT-antigen was isolated more frequently (25.5%) from girls with uterine cervicitis than from those without inflammation (5.8%) (p less than 0.001). Younger girls seemed to be more susceptible to chlamydial infection, because CT-antigen was detected at a higher rate (21.1%) in the 15-17 year age group than in the 18-20 year group (4.6%) (p less than 0.01). Anti-CT IgG antibody (CT-antibody) was found in 65 sera (46.1%). CT-antigen was detected more frequently in CT-antibody positive girls than in those with negative tests (p less than 0.001). Such results suggest that serum antibody does not effectively prevent chlamydial infections in the genital tract. The duration of persistent chlamydial infection was estimated to be from 41 to 367 days, based on the period between the last intercourse and the latest CT-antigen positive day. CT-antigen disappeared spontaneously in some girls (9 of 15) during a 100 day period of abstinence.

C. trachomatis isolation by culture method using McCoy cells and direct immunofluorescence test (DFA) were carried out on specimens collected from patients with suspected chlamydial infections and the results were compared. Follow-up survey was also made on C. trachomatis positive patients. The results are summarized as follows: 1) One thousand and eighty-six specimens were examined. Positive-coincidence ratio obtained from both methods was 80% and negative-coincidence ratio was 92%. The antigen detection by the DFA exceeded a little bit in positive ratio than that by the culture method. 2) C. trachomatis was positively detected in 31 of 94 men (33%) and 5 of 27 (17%) women during periods from 7 days to 2.5 years in follow-up examinations.

Sixty-one male urethritis cases, 28 gonococcal urethritis and 33 nongonococcal urethritis (NGU), were out-patients at the Department of Urology, Asahi General Hospital, during the 4 months, Oct. 1, 1984-Jan. 31, 1985. Thirteen of the 33 NGU patients (39.4%) were infected with C. trachomatis (CT). The efficacy of Doxycycline and the route of infection was studied in the cases of CT-positive CT-negative-NGU. CT infection from prostitutes was not so frequent as in the other pathogen infection of NGU. The efficacy of Doxycycline (100 mg b.i.d. for 2 weeks) against CT infection was excellent in the disappearance of subjective complaints and that of white blood cells in the urethral discharge in the CT positive-NGU group (13/13), in comparison with CT negative-NGU group (7/18). The efficacy of Doxycycline against CT was also confirmed from the follow-up study by the isolation of CT and by detection of CT antigen from urethral swabs using FITC conjugated monoclonal antibody against CT antigen.

Between January, 1984 and August 1986, we examined 151 male patients with urogenital infections including 114 cases of urethritis, 18 cases of epididymitis, 19 cases of prostatitis at the outpatient clinic of the department of urology, Toho University's Ohmori Hospital. The positive rate for Chlamydia trachomatis from the urethra based on cell cultures was 29.0%, (30.5% for urethritis, 27.8% for epididymitis, and 21.1% for prostatitis). The positive rate for non-gonorrheal urethritis was 33.0% and chlamydia infections were complicated by gonorrheal urethritis in 23.3% of the cases. The mean age of the patients with positive Chlamydia trachomatis was 31.9 years. The route of infection in 77.8% of the cases was sexual intercourse the most common cause of which was infection from a prostitute in 62.1% of the cases. The latent period was in most cases from 2-3 weeks.

During the past 3 years, 30 sexual partners including 18 married couples and 12 extramarital sexual pairs whose male partners were diagnosed as having non-gonococcal urethritis were examined for Chlamydia trachomatis infection. Twenty-three of the 30 couples (76.7%) had identical results either positive or negative for Chlamydia trachomatis infection. All 3 male partners of the 3 pairs who had the non-identical results for Chlamydia trachomatis infection, male negative and female positive, had history of urethritis or prostatitis. Fourteen of the 17 female partners (82.4%) who were positive for Chlamydia trachomatis, had no subjective complaints. The above findings suggest the necessity of treating the female sexual partners of the non-gonococcal urethritis patients irrespective of their symptom.

Enoxacin (ENX) is a new derivative of pyridonecarboxylic acid and has a broad spectrum antibacterial activity. In this study, the effects of ENX on nongonococcal urethritis (NGU) were examined in 20 patients. ENX was administered orally 3 times a day at a daily dose of 600 mg for 2-23 days. Before treatment, Chlamydia trachomatis was isolated from urethral epithelium in 5 of the 20 patients. Of the 18 patients evaluated in this clinical trial, 15 (83%) responded well, and ENX was effective in 3 of the 5 Chlamydia positive cases and in 12 of the 13 Chlamydia-negative patients. As adverse reactions, headache was observed in one case and numbness of limbs in another case. No abnormality induced by ENX was recognized in laboratory findings. It is concluded that ENX is a useful drug in the treatment of patients with NGU, particularly in whom Chlamydia trachomatis is not isolated.

From January through December 1986, the urethral smear specimens from 132 male urethritis patients were examined by using Chlamydia trachomatis direct specimen test (Micro Trak). C. trachomatis was detected in 59 (44.7%) out of 132 male urethritis patients, 8 (30.8%) out of 26 patients with gonococcal urethritis (GU), and 51 (48.1%) out of 106 patients with non-gonococcal urethritis (NGU). In 5 (31.3%) out of 16 cases, already treated in other hospitals, infection of C. trachomatis was revealed. The age distribution of the patients with chlamydial urethritis (NGU-C) was between 19 and 52 years old, and the average was 30.9 years old. The peak incubation periods of GU and NGU-C were seen within 7 days and between 8 and 14 days, respectively. In NGU-C patients, 70.6% of them showed WBC less than or equal to 9/hpf in first voided urine sediment. The source of infection was a non-prostitute in 22.2% and a prostitute in 72.2% of GU, while in NGU-C 23.5% was infected from a non-prostitute, and 62.7% from a prostitute. Ofloxacin (OFLX) was administered in a daily dosage of 600 mg (in 3 divided oral doses) for 5 to 14 days in 38 NGU-C patients, and C. trachomatis was eliminated in all cases after the treatment.