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Mycoplasma genitalium is one of the pathogenic microorganisms in male urethritis as a sexually transmitted infection (STI). M.genitalium is detected in the urine specimens of 15-25% male patients with urethritis. The emergence of macrolide- or fluoroquinolone-resistant M.genitalium has become a serious problem in the treatment of male urethritis worldwide, but there is no commercial-based detecting kits accepted by the national insurance in Japan. In this study, we tested the validity of a molecular kit for detecting seven microorganisms related to STI (Anyplex™ II STI-7 Detection which detects Neisseria gonorrhoeae, Chlamydia trachomatis, M.genitalium, Mycoplasma hominis, Ureaplasma urealyticum, Ureaplasma parvum, Trichomonas vaginalis) produced by Seegene company in Korea. Seventeen M.genitalium strains were used to determine the detection limit of M.genitalium. M.genitalium DNA samples were extracted from M.genitalium strains and the diluted DNA samples were reacted to detect M.genitalium by the Anyplex™ II STI-7 Detection. The detection limit was determined as the maximum dilution of DNA samples and the number of M.genitalium DNA copies calculated. In this study, the minimum DNA copies to detect M.genitalium by the Anyplex™ II STI-7 Detection was determined to be around 50 per reaction. The detection rates of M.genitalium in urine specimens were compared between MgPa gene PCR and the Anyplex™ II STI-7 Detection. The positive and negative concordant rates were high as 96.4% (27/28) and 98.6% (71/72), respectively. The validity of the kit for detecting seven microorganisms related to STI (Anyplex™ II STI-7 Detection) was high and thought to be useful for clinical uses.

Six regional medical associations in Shiga prefecture agreed to cooperate in an investigation of medical care for male gonococcal and chlamydial urethritis. In June 2011, we sent a questionnaire to 372 medical offices in Shiga prefecture, and analyzed replies of respondents. Ten urologists and 175 non-urologists responded to the survey (response rate 49.7%). Among 185 physicians, 52 (10 urologists and 42 nonurologists) have treated male patients with gonococcal and chlamydial urethritis. More than 20% (42/175) of non-urological clinics are involved in the medical management. At initial diagnosis for sexually transmitted male urethritis, all urologists select the nucleic acid amplification method (100%), whereas many non-urologists do not (35%). For the treatment of chlamydial urethritis, non-urologists select levofloxacin (LVFX, 52.8%) rather than azithromycin (AZM, 22.0%), whereas urologists use AZM (78.0%) mostly but do not use LVFX (0%) (p = 0.023). For the treatment of gonococcal urethritis, non-urologists prefer oral new quinolones (53.1%) compared to urologists (25.0%) (p = 0. 74). For cure judgment of gonoccocal and chlamydial urethritis, many non-urologists rely on the improvement of subjective symptoms (50 and 47%), but urologists do not (10 and 0%) (p = 0.022 and 0.026, respectively). As for recognition of the clinical guideline for sexually transmitted disease, most urologists (90%) know it, but few non-urologists (13%) do (p < 0.001). We found that non-urological clinics make a great contribution to the medical treatment for male gonococcal and chlamydial urethritis in Shiga prefecture. It is important to standardize the medical care for sexually transmitted male urethritis by familiarizing non-urological practitioners with the clinical guideline.

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.

Besides Chlamydia trachomatis, various microorganisms could cause non-gonococcal urethritis (NGU). Recently, Mycoplasma genitalium and Ureaplasma urealyticum (biovar 2) have been suggested to be other pathogens of NGU independent of C. trachomatis. Clinical findings of non-chlamydial NGU, including M. genitalium--or U. urealyticum-postive NGU, are not different from those of chlamydial NGU. M. genitalium and U. urealyticum (biovar 2) are susceptible to tetracyclines, macrolides, and fluoroquinolones. However, the post-treatment presence of M. genitalium in the urethra is significantly associated with persistent or recurrent urethritis. Eradication of this mycoplasma from the urethra is essential for managing M. genitalium-positive NGU. In treatment of non-chlamydial NGU, therefore, the antimicrobial agents that are active against M. genitalium should be chosen.

Genital chlamydial infection is a sexually transmitted disease (STD) of public health importance. Infection with Chlamydia trachomatis can cause urethritis, cervicitis, pharyngitis, or epididymitis, although asymptomatic infections are quite common. Chlamydial infection remains asymptomatic in approximately 50% of infected men and 70% of infected women. Genital infection with C. trachomatis markedly enhances the risk for reproductive tract sequelae in women, including tubal factor infertility and ectopic pregnancy. A proper detection of C. trachomatis and its treatment is of importance to prevent outbreaks of genital chlamydial infection. We review the pathology, diagnosis and treatment of this entity.

Sexually transmitted diseases diagnosed and treated exclusively in urology clinics are male urethritis and epididymitis caused by Neisseria gonorrhoeae and Chlamydia trachomatis. Although recent developments in diagnostic tools have made it possible to detect these microorganisms with extremely high sensitivities, the screening of Neisseria gonorrhoeae using the microscopic observation of intraleucocyte diplococci stained by methylene blue or gram staining in the first voided urine sediment or urethral smears still the key point in the diagnosis and treatment of gonococcal urethritis. In recent years, mycoplasmas and ureaplasmas have been supposed to be possible pathogens of prostatitis as well as urethritis. The objective diagnosis of genital skin lesions caused by herpes simplex virus and human papillomavirus are remained to be developed.

In Japan, Neisseria meningitidis is not sufficiently recognized as the primary causative bacteria of sexually transmitted diseases (STDs) as the number of reported cases is small. Here, we summarize reports from 3 medical institutions, present clinical courses for each case, as well recommending precautions to prevent infection with this bacterium. Fourteen cases of N. meningitidis urethritis (MU) were admitted between April 2001 and June 2006. All patients were male, consulted a doctor after experiencing subjective symptoms, such as micturition pain and pus discharge, and were diagnosed as having urethritis using isolation culture methods. In 8 of the 14 cases, history of sexual contact in the preclinical stage was confirmed, and contact was with a commercial sex worker (CSW) in 6 of these cases. Many of these patients recalled oral contact. All strains indicated susceptibility to many drugs, and there were no problems with treatment. With regard to serotype, there were 10 cases of type Y, 1 case of type B, and 3 cases that were not classifiable or unidentified. In addition, among the 9 strains that were subjected to genotype identification, 7 strains were ST-23. The recent increase in availability of nucleic acid amplification methods has facilitated simultaneous detection of Neisseria gonorrhoeae and Chlamydia trachomatis. However, we fear that MU will become latent. For screening of urethritis, Gram staining and culture of urethral material must be performed to detect this disease. The relationship of the detected strain and its role in the pathogenesis of meningitis are uncertain, but its serotype and genotype are common in cases of meningitis. Thus, precautions are required to prevent spread of this bacterium.

A 16-year-old heterosexual man presented to our hospital with a purulent urethral discharge and pain at voiding. These symptoms began seven days after oral-genital contact (fellatio) with his partner. A Gram-stained smear from the urethral discharge showed Gram-negative diplococci, and the antigen of Chlamydia trachomatis from urine was positive. We initially made a diagnosis of urethritis caused by Neisseria gonorrhoeae and C. trachomatis. However, N. meningitidis was isolated by culture. Clinicians should pay attention to the possibility of N. meningitidis infection in all cases resembling gonococcal urethritis.

We reviewed the results of clinical examinations conducted on and antimicrobial susceptibilities to Neisseria gonorrhoeae isolated from 51 patients with gonococcal urethritis who visited our hospital during the period from February 2005 to April 2006. The type of sexual activity by which the bacteria was transmitted was oral sex in 81.6% (40/49) of the patients, and none of the patients used a condom during oral sex. Fifty percent (24/48) of the patients were aware of the risk of getting sexually transmitted diseases (STDs) from having oral sex without the use of a condom. The ratios of drug-resistant N. gonorrhoeae were 56.9% for Penicillin G, 0% for CVA/Amoxicillin, Azithromycin and Minocycline, 5.9% for Cefpodoxime, 2.0% for Ceftriaxone, 0% for Cefodizime, and 84.4% for Ciprofloxacin, Levofloxacin and Gatifloxacin. The bacteria in 7.8% of the cases were beta-lactamase-producing strains. The results of this study indicated that STD from oral sex is prevalent in Sapporo. Increasing resistance to Quinolone was noted, but levels of susceptibility to other drugs remained relatively high.

We report a case of acute spongiositis with diabetes mellitus. A 63-year-old man with a 7-year history of diabetes mellitus was admitted to our hospital complaining of perineal pain and pyrexia. Physical examinations revealed induration and tenderness of the corpus spongiosum, but no signs of rubor or erosion. A laboratory investigation showed leukocytosis (12,000/microliter), an elevated level of C-reactive protein (3.38 mg/dl) and hyperglycemia (532 mg/dl). Pyuria and bacteriuria were absent. Magnetic resonance imaging (MRI) yielded abscess-like lesion of the corpus spongiosum. Culture of the aspirated specimen demonstrated B-hemolytic streptococci. Treatment with antibiotics and insulin resolved leukocytosis and tenderness of corpus spongiosum. He left the hospital on the 23rd day after admission and no evidence of recurrence of the spongiositis was found for 1 year after discharge.

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.

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.

For 10 strains of Neisseria gonorrhoeae exhibiting decreased susceptibility to new quinolones (MIC, > or = 0.1 microgram/ml) isolated and preserved from patients with gonococcal urethritis during the period from February 1991 through January 1992. We investigated the mechanisms for development of resistance to new quinolones. After selecting 3 PCG-sensitive and new quinolone-resistant strains from among these strains, we first carried out transformation experiments of N. gonorrhoeae using a plasmid carrying the E. coli gyrA gene. We then determined the base sequence on the N. gonorrhoeae gyrA gene by PCR method. Of the 3 strains in which transformation experiments were carried out, 2 strains yielded transformants, one of which was 8 times more sensitive to norfloxacin (NFLX) than the original strain, and it was assumed that this strain has a mutation in the gyrA gene. In our study of the base sequence on the N. gonorrhoeae gyrA gene using the PCR method, both strains had the mutation of Ser 83 (TCC)-->Phe (TTC), and in addition to this mutation of Ser-->Phe, one other strain had the mutation of Asp-87 (GAC)-->Gly (GGC). This type of mutation of quinolone-resistant N. gonorrhoeae on the gyrA gene has high homology with the mutations reported for Escherichia coli and other strains and there appears to be a close correlation between the increasing frequency of use of the new quinolones in this country and the spread of such resistant strains of N. gonorrhoeae.

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.

Epidemiological studies on sexually transmitted diseases (STDs) in Saitama Prefecture, from 1989 through 1991, were performed and the following results were obtained. The number of patients was 1,833 (male: 1,532, female: 30) cases in 1989, 1,294 (male: 1,163, female: 131) cases in 1990, and 1,672 (male: 1,478, female: 149) cases in 1991. Male patients in their twenties or thirties were often affected by STDs. The order of frequency of STDs was roughly as follows; non-gonococcal and non-chlamydial urethritis, gonococcal urethritis, chlamydial rethritis, condyloma acuminatum, genital herpeic infection and syphilis. Concerning the criteria of the diagnosis on chlamydial urethritis, cases diagnosed by only clinical findings decreased markedly from 1989 through 1991, cases diagnosed by the combination of the antibody of C. trachomatis and clinical findings increased, and cases diagnosed by the detection of C. trachomatis were always about 70%. Male patients were mostly infected from prostitutes. Many patients with STDs were often infected in Taiwan, the Philippines, the Kingdom of Thailand and Republic of Korea.

A polymerase chain reaction (PCR) method was compared to standard methods (cultures for Neisseria gonorrhoeae and Chlamydia trachomatis and an enzyme-immunoassay for C. trachomatis) in diagnosis of gonococcal and chlamydial urethritis in 40 male patients with urethritis. Gonococcal urethritis was diagnosed by detection of a 206 bp DNA fragment amplified by PCR with N. gonorrhoeae-specific primers. Chlamydial urethritis was diagnosed by detection of a 242 bp DNA fragment amplified by PCR with C. trachomatis-specific primers. Gonococcal and chlamydial urethritis, gonococcal and non-chlamydial urethritis, non-gonococcal and chlamydial urethritis, and non-gonococcal and non-chlamydial urethritis were diagnosed in 8, 10, 14 and 8 patients, respectively, by the PCR method. In 9 patients with gonococcal and chlamydial urethritis, 10 with gonococcal and non-chlamydial urethritis, 12 with non-gonococcal and chlamydial urethritis, and 9 with non-gonococcal and non-chlamydial urethritis, diagnosed by the standard methods, the coincidence rates of the PCR to the standard methods were 78% (7/9), 90% (9/10), 100% (12/12), and 89% (8/9), respectively. The overall coincidence rate between the PCR and the standard methods in diagnosis of urethritis were high (90%). In addition, N.gonorrhoeae and C.trachomatis could be simultaneously detected from one urethral sample in approximately 6 hours by means of the PCR. Thus, the PCR method could clinically be applied and would offer several advantages to diagnosis of urethritis, compared to the standard methods.

A polymerase chain reaction (PCR) procedure for detection of Ureaplasma urealyticum was developed. A set of oligonucreotides based on sequences within the 16S ribosomal RNA gene from U. urealyticum were used as extension primers for the PCR. A DNA fragment of 397 bp was amplified by the PCR, when U. urealyticum DNA was template for the PCR. No amplified product was detected from other bacterial DNA including those of Mycoplasma genus. The amplified DNA fragment of 397 bp was detected on agarose gel electrophoresis, when DNA of > or = 10(2) cells of U. urealyticum per PCR was used as template for the PCR. Thus, the PCR procedure was shown to be a simple, rapid and specific method for detection of U. urealyticum and could be applied to detection of U. urealyticum from clinical specimens.

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.

Epidemiological and bacteriological studies on Neisseria gonorrhoeae isolated in Sapporo, Japan, in 1980 and 1991 performed and the following results were obtained. 1. The range of age in the patients infected with Neisseria gonorrhoeae tended to be younger than those in the whole country. 2. Male patients in the early 20s or younger with gonococcal urethritis were often infected by bon-professional females but those in their late 20s or older were often infected from professional females, for example prostitutes and hostesses. 3. The rate of professional females who were positive to gonococci reached 17.4% and young females in their teens with cervicitis had the highest morbidity rate of gonococci than those in the older females. 4. The latent period in gonococcal infections tended to become longer gradually. 5. The isolation rate of penicillinase producing Neisseria gonorrhoeae (PPNG) showed a peak of 23.9% (61/255) in 1985, but gradually declined thereafter and it was 3.7% (1/27) in 1991. 6. An investigation on auxotype showed a decline of proto and Pro-strains and an increase of AHU-strains in non-PPNG. And most of the PPNG belonged to proto or Pro-strains. 7. With the relationship between auxotype and sensitivity to AMPC, AHU-strains were more sensitive than proto or Pro-strains.

A polymerase chain reaction (PCR) procedure was developed for detection of Neisseria gonorrhoeae. Two oligonucleotides based on sequences within a 16S ribosomal RNA gene from N. gonorrhoeae were used as extension primers for the PCR. A single DNA fragment of 206 bp was amplified, when N. gonorrhoeae DNA was template for the PCR. No amplified product was detected in Chlamydia trachomatis DNA, Ureaplasma urealyticum DNA or other bacterial DNAs. The DNA fragment of 206 bp was detected on agarose gel electrophoresis, when DNA of greater than or equal to 6.5 N. gonorrhoeae per PCR was used as template DNA for the PCR. The culture and the PCR were carried out for detection of N. gonorrhoeae in 67 urethral swabs obtained from male patients with urethritis. In 27 of 28 specimens in which N. gonorrhoeae was isolated and identified by the culture, 206 bp DNA fragment was amplified by the PCR, but in one specimen no DNA fragment was detected. In 2 of 39 culture-negative specimens, 206 pb DNA fragment was detected and in the remaining specimens, PCR was negative for N. gonorrhoeae. The overall detection coincidence rate between the culture and the PCR was 95.5% (64/67). Thus, the PCR procedure developed in this study was sensitive and specific for detection of N. gonorrhoeae and could be applied for diagnosis of gonococcal urethritis.

We performed basic and clinical studies in male gonococcal urethritis on a new oral antimicrobial agent, levofloxacin (LVFX, DR-3355), a new quinolone derivative. The antibacterial activity of LVFX against clinical strains of Neisseria gonorrhoeae was roughly comparable to that of ofloxacin, and ciprofloxacin. LVFX was administered to 10 males with gonococcal urethritis, 200 mg twice a day (8 cases) or 100 mg twice a day for 3 days (2 cases). Clinical evaluation was made according to the criteria of the Japanese UTI Committee. Overall efficacy rate was 100% (10/10). In the complication of chlamydia trachomatis (2 cases), efficacy rate was 100% (2/2). No subjective or objective adverse reactions occurred.

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.

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.

Results of the treatment of 5 cases of males with uncomplicated gonoccocal urethritis using levofloxacin (LVFX, DR-3355), the L-type optical isomer of ofloxacin (OFLX), were compared with those treated with OFLX itself. Three hundred mg/day of LVFX or 600 mg/day of OFLX was given to each patient for 5 days. Both drugs showed excellent clinical results in all the patients. When MICs of the 2 drugs were compared against 57 isolated strains of Neisseria gonorrhoeae including 3 penicillinase-producing N. gonorrhoeae, it was found that MICs of LVFX were approximately one half of those of OFLX.

Ureaplasma urealyticum was investigated in urine from 765 outpatients who visited Jikei University Affiliated Hospital and Tokyo Metropolitan Taito Hospital from June, 1988 to December, 1989 in order to clarify the pathogenicity of U. urealyticum in human genitourinary tract. U. urealyticum in urine was detected by means of Taylor-Robinson's method. The positive rates of U. urealyticum were 31.5% in 146 patients with gonococcal urethritis, 33.8% in 334 patients with non-gonococcal urethritis, 17.5% in non-bacterial chronic prostatitis and 27.5% in the other patients without infectious diseases, respectively; no significant difference was seen among these groups. U. urealyticum was detected in the urine from 32.1% of the 28% patients who were younger than 12. However, U. urealyticum was detected in the urine from 5.6% of the 18 patients who were older than 70. Therefore, there was no relationship between the age and U. urealyticum-positive rate in urine. Furthermore, there was no relationship between the detection of U. urealyticum and the subjective and objective findings in the patients with urethritis before and after the treatment. From these results, it is presumed that U. urealyticum has no pathogenicity in human genitourinary tract.

We conducted a multi-center clinical study to evaluate the role of Ureaplasma urealyticum and Mycoplasma hominis in male urethritis. The incidence of each organism in first-voided urine samples of 160 male urethritis patients, including 28 with gonococcal and 126 with non-gonococcal urethritis, was investigated. U. urealyticum and M. hominis were isolated from 13.6% and 6.5%, respectively, of the urine samples, and the concentration of each mycoplasma exceeded 10(3) ccu/ml (color changing units/ml) in 5.2% and 3.9%, respectively. Among 64 patients with non-gonococcal non-chlamydial urethritis, U. urealyticum and M. hominis concentration exceeding more than 10(3) ccu/ml were detected in only two and one samples, respectively. The incidence among urethritis patients with a concentration of either mycoplasma exceeding 10(3) ccu/ml was not significantly greater than that among subfertile males without urethritis. The findings of the present study suggest that, although mycoplasmas may cause urethritis in some patients, the incidence of urethritis due to U. urealyticum or M. hominis is low among patients with non-gonococcal, non-chlamydial urethritis.

We performed basic and clinical studies on the effects of a new oral antimicrobial agent, fleroxacin (FLRX), a new quinolone derivative in male gonococcal urethritis. The antibacterial activity of FLRX against clinical strains of Neisseria gonorrhoeae was roughly comparable to that of norfloxacin and ofloxacin. FLRX was administered to 58 males with gonococcal urethritis. Two different schedules of administration were adopted. One was a single-dose of 300 mg given orally (17 cases) and the other was the oral administration of 200 mg once a day for 3 to 10 days (41 cases). Clinical evaluation was made according to the criteria of the Japanese UTI Committee. The overall efficacy rate was 98% (49/50). For complications of Chlamydia trachomatis (11 cases), the efficacy rate was 90.9% (10/11). No subjective or objective adverse reaction occurred.

We reviewed 497 patients with male urethritis diagnosed between January, 1986 and March, 1989 at the Asama General Hospital. The incidence of gonococcal urethritis (GU) was 47.7%, and that of non-gonococcal urethritis (NGU) 52.3%. There was no difference in the age distribution between GU and NGU. Prostitutes were the most common source of the infection in both GU and NGU. Incubation periods were longer in NGU than in GU, statistically. Urethral discharge was the most common symptom. Purulent urethral discharge was seen more commonly than serous urethral discharge in GU. On the contrary, serous urethral discharge was more common in NGU. Penicillin-resistant gonococcus comprised 29.4% and mixed infection of the C. trachomatis existed 25.6% in GU. C. trachomatis was detected in 71.8% in NGU. In GU, new quinolones and penicillins were administered frequently. The effective rates 1 week after the administration were 80.6% and 83.3%, respectively. In NGU, new quinolones and minocycline were administered frequently. The effective rates were 70.4% and 85.3%, respectively. Ofloxacin (OFLX) showed the highest effective rate to NGU among the four new quinolones. The relapse rate for the two-week administration group was lower than that for the one-week-administration group, but the difference was not statistically significant.

We evaluated the clinical usefulness of a new EIA kit using a monoclonal antibody, IDEIA CHLAMYDIA (IDEIA, Novo Nordisk), for detection of C. trachomatis antigen from the genital tracts of male and female cases. The results were compared with those by Chlamydiazyme (Abbott). 1. C. trachomatis antigen detection by the IDEIA and Chlamydiazyme tests before treatment; IDEIA has a significantly higher detection rate (38.0%, 105/276) than Chlamydiazyme (29.8%, 80/276), for C. trachomatis antigen from urethral smears of 276 male patients with urethritis. In 646 female cases, including cervicitis and so on, IDEIA detected C. trachomatis antigen from cervical smears in 14.5% (94/648) of the total, while Chlamydiazyme did so in 11.9% (77/648). When considering the different results using IDEIA and Chlamydiazyme, approximately 20% of the IDEIA-positive cases were Chlamydiazyme-negative. However, when IDEIA was negative, less than 1% showed Chlamydia-positive. 2. C. trachomatis antigen detection during and after treatment; We studied the clinical courses of 14 male urethritis and 8 female cervicitis cases who had had positive results with both IDEIA and Chlamydiazyme before treatment. Two of the 14 urethritis cases showed positive results with IDEIA, but not with Chlamydiazyme after either 7 or 14 days treatment by an antimicrobial agent. These two also had symptoms indicating persistent urethritis. One of the 8 female cervicitis cases showed a positive result with IDEIA but not with Chlamydiazyme after 7 days treatment by an antimicrobial agent, and this case also had symptoms indicating persistent cervicitis. Thus, these clinical findings suggest that IDEIA can detect even a small quantity of antigen soon after treatment, but Chlamydiazyme can not. In conclusion, IDEIA has a higher sensitivity than Chlamydiazyme, in the detection of C. trachomatis antigen, suggesting that IDEIA is more useful.

The detection of C. trachomatis antigen in first-voided urine sediments has recently been achieved by means of IDEIA CHLAMYDIA (IDEIA, Novo Nordisk), an EIA kit using monoclonal antibodies. Therefore, this kit was used as a screening test to examine the infection rate of C. trachomatis in young adult men without symptoms. The titers of serum IgA and IgG antibodies to C. trachomatis were also determined. 1. Antigen detection from first-voided urine sediments of young adult men without urogenital symptoms (141 cases): The detection rate by IDEIA was 5.0% (7/141). Three of the 7 cases which were positive for antigen in first-voided urine sediments were reaffirmed as having asymptomatic C. trachomatis urethritis, since they were also revealed to have C. trachomatis in the urethra. 2. The positive rates of serum antibodies: The titers of serum antibody were determined in 128 cases out of the 141 cases. The positive rates of IgA and IgG were 6.3% (8/128) and 35.9% (46/128) respectively. The positive rates of IgA and IgG antibodies were significantly higher in cases with positive antigen in first-voided urine sediments than in those which were negative. These results indicate that this kit is useful for antigen detection. 3. The screening test revealed asymptomatic C. trachomatis infections in 5% of young adult men, suggesting the extensive spread of the infection. The screening test using first-voided urine sediments will be useful in public health.

The Plasmid DNA patterns of 22 penicillinase-producing Neisseria gonorrhoeae (PPNG) strains isolated from male patients with gonococcal urethritis in 1988 at Tokyo Metropolitan Taito Hospital were determined. The 3.3 megadalton plasmid, the so called the African type plasmid, was present in 3 isolates out of 22 PPNGs, all of which occurred with a conjugative 24.5 megadalton plasmid. 4.4 megadalton plasmid was present in 19 isolates out of 22 PPNGs, 5 of which occurred with a conjugative 24.5 megadalton plasmid. The infective sources of the 3 cases of the African type PPNGs were found as 1 from philippines and 2 from prostitutes, so called Soap Land Girl in Japan. And they suggests that the African type PPNGs are already common in Japan.

AT-2266 tablets were administered orally in a daily dosage of 600 mg for 7 consecutive days as treatment of non-gonorrheal urethritis in 106 cases. The therapeutic results in 90 cases were investigated, and a detailed analysis of various background factors was also performed. Most of the patients were between 15 and 35 years old; 11.3% of the patients were between 15 and 19 years old. The source of infection was a "professional" woman such as a prostitute in 37 cases, and a non-professional woman in 53 cases. This is noteworthy because the source is more often a "professional" women in the case of gonorrhea. Bacterial isolates were obtained by culture of the urethral secretions. Most were gram-positive cocci, especially S. epidermidis, but, in 51.7% of the patients, no bacteria could be cultured. The urethral secretion was purulent in 51.9% of the patients, and serous in the remaining 48.1%. The efficacy rates obtained when AT-2266 was administered in a daily dosage of 600 mg (in 3 divided oral doses) for 7 days were as follows. In the purulent secretion group, the 3-day and 7-day therapeutic efficacy (elimination of the secretion) rates were 24.2% and 61.2%. In the serous secretion group, the corresponding efficacy rates were 36% and 71.4%. Thus the therapeutic efficacy was slightly inferior in the purulent secretion group. Oral administration of AT-2266 resulted in side effects in 5 of the patients (5.5%). These side effects consisted primarily of mild gastrointestinal disturbances, which did not require discontinued drug administration.

The in vitro activity of minocycline (MINO) against Chlamydia trachomatis and its efficacy in the treatment of C. trachomatis-associated nongonococcal urethritis were investigated. Six isolates of C. trachomatis were inhibited at 0.06 micrograms/ml of MINO and 5 isolates at 0.03 micrograms/ml. All cases received oral MINO twice daily for 7 or more days in doses of 100 mg. In 5 of 31 cases, 2 g of spectinomycin was intramuscularly administrated together with MINO only once. C. trachomatis was eliminated in all cases tested. Excellent results were obtained in 26 cases (84%); urethral discharge and polymorphonuclear cells (PMN) disappeared or decreased to normal levels (3 cells/hpf or less) in these cases. Ureaplasma urealyticum was isolated from 8 cases, 7 of which became free of ureaplasmal infection. MINO seemed to be less effective on the decrease of PMN in the urethral smear in cases infected coincidentally with C. trachomatis and U. urealyticum than in cases infected with C. trachomatis alone. No subjective side effects were observed in any of the 31 cases studied. In conclusion, MINO was a useful antimicrobial agent for the treatment of C. trachomatis- and U. urealyticum-associated nongonococcal urethritis.

Thirty male patients with gonorrheal urethritis were treated with azthreonam at the urological ward of Tokyo Metropolitan Taito Hospital during the period from January to March, 1984, and clinically evaluated. Sixty-one strains of Neisseria gonorrhoeae isolated from the clinical specimens including those of the patients were bacteriologically studied. Of the 61 strains, 9 (15%) were PPNG strains and the other 52 strains were non-PPNG strains. MICs of PCG to the 9 PPNG strains were 3.13--50 micrograms/ml and to the 52 non-PPNG strains were 0.024--3.13 micrograms/ml. While MICs of azthreonam to the PPNG strains were 0.024--0.20 micrograms/ml and to the non-PPNG strains were 0.0122--0.78 micrograms/ml. Those patients with gonorrheal urethritis were given a single 1 g intramuscular dose of azthreonam. The following clinical findings were obtained: The clinical efficacy rate of 30 cases was 100%; excellent in 16, good in 14 cases. All 2 patients with gonorrheal urethritis caused by PPNGs were cured with the treatment. No subjective side effects were found.

Thirty male patients with gonorrheal urethritis were treated with norfloxacin (NFLX) at our urological ward between May and July, 1984, and clinically evaluated. Thirty-nine strains of Neisseria gonorrhoeae isolated from the clinical specimens of these patients were bacteriologically studied. Of the 39 strains, 3 (7.7%) were PPNG strains and the other 36 strains were non-PPNG strains. The minimum inhibitory concentration (MIC) of penicillin G, PCG to the 3 PPNG strains was 6.25-25 micrograms/ml and that to the 36 non-PPNG strains was 0.05-3.13 micrograms/ml. The MIC of NFLX to the PPNG strains was 0.025-0.1 microgram/ml and that to the non-PPNG strains was less than or equal to 0.0125-0.2 microgram/ml. The patients with gonorrheal urethritis were given NFLX orally for 3 days at a daily dose of 600 mg. The clinical efficacy rate was 100%: excellent in 12 and good in 18 cases. All 3 patients with gonorrheal urethritis caused by PPNGs were cured. No subjective side effects were found.

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.

Forty-one male patients with gonorrheal urethritis were treated with ceftizoxime (CZX) administered concomitantly probenecid at the urological ward of Tokyo Metropolitan Taito Hospital during the period from October, 1982 to February, 1983, and clinically evaluated. Eighty strains of Neisseria gonorrhoeae isolated from the clinical specimens of these patients were bacteriologically studied. Of the 80 strains, 12(15%) were PPNG strains and the other 68 strains were non-PPNG strains. MICs of PCG to the 12 PPNG strains were 3.13 - 25 micrograms/ml and to the 68 non-PPNG strains were 0.0122 - 3.13 micrograms/ml. While MICs of CZX to the PPNG strains were less than or equal to 0.003 - 0.024 micrograms/ml and to the non-PPNG strains were less than or equal to 0.003 - 0.05 micrograms/ml. Those patients with gonorrheal urethritis were given a single 0.5 g intramuscular dose of CZX and a single 1 g oral dose of probenecid. The following clinical findings were obtained: The clinical efficacy rate of 41 cases was 98%; excellent in 24, good in 16, poor in 1 case. All 3 patients with gonorrheal urethritis caused by PPNGs were cured with the treatment. No subjective side effects were found.

Eighty-nine isolates of Neisseria gonorrhoeae from 89 male urethritis patients were studied for beta-lactamases (penicillinase) production, and their susceptibilities to benzylpenicillin, amoxicillin, piperacillin, clavulanic acid/amoxicillin (Augmentin), cephalexin, cefotaxime, spectinomycin and minocycline were determined by an agar plate-dilution method. Penicillinase activity was tested by a chromogenic cephalosporin method with nitrocefin as substrate (CefinaseTM discs, BBL, USA) and by a paper strip acidimetric method with benzylpenicillin as substrate (beta-Lactamase detection papers, Oxoid, UK). In addition, 60 of the 89 patients were examined for Chlamydia trachomatis, using fluorescein-labeled monoclonal antibodies (Direct specimen test; Micro TrakTM, Syva Co., USA). Penicillinase-producing N. gonorrhoeae (PPNG) were found in 12 of the 89 strains (13.5%). Although all these strains of PPNG were highly resistant to benzylpenicillin and amoxicillin, the minimum inhibitory concentrations (MICs) of Augmentin markedly decreased. Piperacillin was highly active against not only non-PPNG but also PPNG strains. More than half the isolates were resistant to cephalexin (MICs greater than or equal to 12.5 micrograms/ml) while all strains including non-PPNG and PPNG were fully sensitive to cefotaxime (MICs less than or equal to 0.20 microgram/ml). Spectinomycin and minocycline had MIC ranges of 6.25 to 25 micrograms/ml and 0.01 to 3.13 micrograms/ml, respectively; the ranges for non-PPNG and PPNG strains were fairly similar. C. trachomatis was detected in 11 (18.3%) out of 60 patients examined. The increasing incidence of PPNG and coexisting chlamydial infection should be taken into account in the treatment of gonococcal urethritis.

Clinical effects of ofloxacin (OFLX) in the treatment of chlamydial urethritis was compared with those in the treatment of non-chlamydial urethritis. Chlamydia trachomatis was isolated from 33 (39.3%) out of 84 patients with nongonococcal urethritis. OFLX was administered at a dose of 100 mg, three times daily (300 mg) for 14 days. In 31 (93.3%) of the 33 patients with chlamydial urethritis, C. trachomatis was eliminated within 7 to 14 days after the start of administration, and, in two patients, inclusion bodies decreased in number but persisted. The subjective and objective clinical symptoms, urethral discharge, polymorphonuclear leucocyte (PMNL) in smears, and pyuria in VB1 of the patients with chlamydial urethritis, disappeared in 56.0, 57.6, and 63.0% of the cases, and improved in 24.0, 30.0 and 29.6% respectively, whereas those of the patients with non-chlamydial urethritis, urethral discharge, PMNL, and pyuria in VB1 disappeared in 57.6, 56.9 and 32.9%, and improved in 26.9, 13.7 and 37.9%, respectively. There was no significant difference in the clinical effects of OFLX between cases of chlamydial urethritis and those of non-chlamydial urethritis. Marked improvement in clinical symptoms were observed between day 7 and day 14 of medication in cases of both chlamydial and non-chlamydial urethritis. In nine out of 11 cases of non-chlamydial urethritis where OFLX was ineffective, no Ureaplasma nor aerobes, nor C. trachomatis, were isolated. In the remaining two cases where Ureaplasma and S. epidermidis were isolated respectively, these two bacteria were eradicated after medication, but urethral discharge and PMNL remained unchanged.(ABSTRACT TRUNCATED AT 250 WORDS)

Fifty six males with gonococcal urethritis were administered 600 mg of norfloxacin (NFLX) daily for 7 days, before the effects were examined. The pharmaceutical effects were studied on the third and the seventh day after administration. Pronounced effectiveness of 77.8%, effectiveness of 19.4% and ineffectiveness of 2.8% were found on the third day after administration, and pronounced effectiveness of 94.4%, effectiveness of 5.6% and ineffectiveness of 0% were found on the seventh day after administration. Effectiveness of 100% was obtained up to the seventh day. Minimum inhibitory concentration (MIC) of NFLX and ampicillin (AMPC) were determined for 14 strains. The peak of MIC of NFLX was found at 0.05 micrograms/ml and all strains were 0.78 micrograms/ml or lower. The peak of MIC of AMPC was 0.1 microgram/ml. MICs of both drugs were 0.78 micrograms/ml or lower, and resistant bacteria were not found. In view of the analyses of age distribution of patients and source of infection, the peak was found in the twenties and the source of infection in 64.7% were, so-called, professional women.