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Isolation of Chlamydia trachomatis from the male urethra.

Chlamydia trachomatis was isolated from 26% of urethral swabs taken from 509 men with urethritis. The highest yield of 68% was obtained from a selected group of men with nonspecific urethritis (NSU) who had a frank urethral discharge. This is a higher than in previous reports, and is significantly higher than the isolation of C. trachomatis from men with less severe urethritis. The higher yield was similar to C. trachomatis isolation rates reported among patients with severe trachoma in hyperendemic areas. Men with a previous history of NSU had low isolation rates. Overall, 30% of 385 men with NSU had positive chlamydial culture results, 7% of 59 men with gonococcal urethritis alone were Chlamydia-positive, 15% of 59 men with gonorrhoea followed by NSU (post-gonococcal urethritis) were Chlamydia-positive, and only 3% of 61 men without urethritis harboured Chlamydia. Swabs taken from the cervical os of 28 of 108 female contacts of men with NSU had a positive result for C. trachomatis. Significantly more pairs of sexual partners had the same chlamydial culture result than had different results. The chlamydial isolation rate was higher among men admitting a casual sexual contact than in men claiming only regular partnerships. The findings provide further evidence for the sexual transmission of C. trachomatis and for its aetiological role in NSU.

Chlamydia Infections

Differences in physicochemical and antigenic properties of chlamydial strains.

Antigenic analysis of Chlamydia psittaci, C. trachomatis, and Lymphogranuloma venereum (LGV) revealed basic physicochemical differences among the three chlamydial strains. These were manifested in structural, isoelectric points, absorption spectra, and in the characteristics of the chromophobe-containing proteins. The effective period of sonication for C. psittaci and C. trachomatis is around 60 min during which the linkages most susceptible to external sonication forces were broken, releasing all attainable cellular components. Denaturation studies demonstrated that less than 50% of protein content of C. psittaci was denatured after 1 h of sonication, only 5% in the case of C. trachomatis. The protein and carbohydrate content of the most reactive fractions in macrophage-spreading inhibition test were different for LGV and C. trachomatis. The structural differences appear to determine the antigenic properties observed among the chlamydial strains as well as the specificity and probably the mechanisms (s) of cellular immune reactivity to Chlamydiae. This in turn may explain the failure of chlamydial vaccines, prepared from stock strains, to protect immunized children against 'wild' chlamydial strains.

Antigens, Bacterial

Spectinomycin hydrochloride in the treatment of gonorrhoea: Its effect on associated Chlamydia trachomatis infections.

Sixty-three heterosexual men were successfully treated with a single injection of spectinomycin hydrochloride 2 g for urethral infections with Neisseria gonorrhoeae. Chlamydia trachomatis was recovered from the urethra of 11 of these men both before and after treatment. In six men, the organism was isolated after but not before treatment. No isolates were obtained from the remaining men either before or after treatment. All 17 of the men who yielded C. trachomatis developed post-gonococcal urethritis. Eight of 46 men from whom no isolate was obtained in their cultures developed post-gonococcal urethritis. Seventeen of 50 women successfully treated with spectinomycin for cervical infections with N. gonorrhoeae yielded isolates of C. trachomatis both before and after treatment. The organism was isolated from five women before but not after treatment, and from four women after but not before treatment. In 24 women culture for C. trachomatis was negative both before and after treatment. Spectinomycin hydrochloride in the dosage used rarely eliminated C. trachomatis from the genital tract of either men or women; in this respect it resembled two other drugs commonly used for the treatment of gonorrhoea-pencillin and ampicillin.

Chlamydia Infections

Chlamydia trachomatis incidence in relation to vaginal microbiota dynamics, immunogenetics and exposures in a cohort of young student women in France.

BACKGROUND: Given the potential role of the vaginal microbiota in the acquisition of Chlamydia trachomatis infections, we aim to investigate its contribution together with immunogenetics and epidemiological exposures to the incidence of C. trachomatis in young women. METHODS: This study involved 313 female students aged 18-24 years from the i-Predict prevention trial in France. Participants provided four self-collected vaginal samples and filled four self-administered questionnaires every 6 months for 18 months. C. trachomatis-positive participants and negative controls with complete follow-up were selected for this analysis and submitted to chlamydia testing and to vaginal microbiota characterization using 16S rRNA amplicon sequencing. Thirteen human single nucleotide polymorphisms (SNPs) related to C. trachomatis susceptibility and severity were also assessed. RESULTS: Compared to 260 non-infected participants, Gardnerella spp., Fannyhessea vaginae and Prevotella timonensis were more abundant in C. trachomatis-incident participants (n=24) before infection. Having a CST IV at the preceding sample compared to a CST I (3.56 [1.08-11.70], p=0.037) was associated with increased risk of C. trachomatis acquisition, as well as having had multiple concomitant partners in the last 6 months (4.33 [1.19-15.72], p=0.028). Lifetime condom use was associated with decreased incidence (OR 0.38 [0.16-0.94], p=0.037). None of the tested human SNPs was associated with C. trachomatis infection. CONCLUSIONS: In this low-risk for C. trachomatis population, having a CST IV-AB vaginal microbiota and associated bacterial anaerobes was a risk factor for C. trachomatis acquisition after adjustment for other exposures. Condom use remains one of the main tools to prevent incidence.

C. trachomatis

Reticulate bodies as single antigen in Chlamydia trachomatis serology with microimmunofluorescence.

Formalin-fixed, purified reticulate bodies (RB) of Chlamydia trachomatis immunotype C/TW-3/OT were used as a serological test antigen in the microimmunofluorescence test. The sensitivity and specificity of the RB antigen were compared to elementary bodies (EB) used as antigens in the detection of C. trachomatis antibodies in human sera by microimmunofluorescence. RB reacted with all known C. trachomatis immunotypes with the same sensitivity as the homotypic EB. In routine serology with sera and endocervical secretions, the RB antigen had a sensitivity similar to that of the EB in detecting serum antibodies, endocervical secretion antibodies, and antibodies of immunoglobulin M and G classes. No false-positive reactions were detected with control sera. All positive reactions showed type-specific antibodies against an EB immunotype. RB seemed to demonstrate chlamydial group reactivity; sera from 10 psittacosis patients diagnosed clinically and serologically by complement fixation showed five positive, three equivocal, and two negative reactions. By immunofluorescence, RB appeared as distinct rings demonstrating uniform peripheral surface fluorescence at their rims. The EB appeared as pinpoint-sized dots. C/TW-3/OT RB used as a single test anitgen should provide a simple and sensitive serological assay for the detection of C. trachomatis antibody.

Antibodies, Bacterial

Chlamydia trachomatis in women: antibody in cervical secretions as a possible indicator of genital infection.

One hundred eighty-five women college students were examined for genital infection with Chlamydia trachomatis. This organism was isolated from nine (5%) of the 185 women. Antibody was demonstrated in the genital secretions of 26 (14%) and in the serum of 70 (38%) of the women. None of the sexually inexperienced women was infected. Among those sexually experienced, the prevalence of isolation of C. trachomatis and of detection of local antibodies in cervical secretions and serum antibodies to C. trachomatis increased in relation to the number of life-time sexual partners. Local antibody appeared to be a more reliable indicator of infection with C. trachomatis than serum antibody in this college population.

Adolescent

Experimental acute salpingitis in grivet monkeys provoked by Chlamydia trachomatis.

Chlamydia trachomatis is a common cause of sexually transmitted diseases. Recently it has been shown that chlamydiae are also responsible for complications to such lower genital tract infections. In this study, isolates of C. trachomatis from the fallopian tubes of patients with acute salpingitis were inoculated direct into the fallopian tubes of two, and through the cervical canal into the uterine cavity of one grivet monkey. The experimental infections resulted in a self-limited acute salpingitis in the three animals. C. trachomatis was recovered from the monkeys 2 and 3 weeks post inoculation. As found at laparotomy, the infected tubes were swollen and reddened, and there was watery exudate in the abdominal ostia. Microscopically, cellular infiltrates--mainly lymphocytes--were seen in the mucosa, muscularis and subserosa of the tubes. Serologically, a primary antibody response with an IgM to IgG conversion was found. Salpingitis did not occur in a control monkey inoculated in the tubes with a medium lacking Chlamydia. The histological changes in the fallopian tubes of the infected monkeys were reminiscent of those described as being characteristic of "gonococcal" salpingitis in man. The fulfilment of Koch's postulates in the animal model used adds to the earlier evidence that C. trachomatis is capable of causing acute salpingitis in humans.

Acute Disease

Infection of the uterine cervix with Chlamydia trachomatis.

For identification of those variables in the history and in the clinical and routine laboratory examination that are most likely to indicate infection of the uterine cervix with Chlamydia trachomatis, 284 women attending a venereal disease clinic were studied, were studied, of whom 58 (20.4%) yielded the organism from the cervix. Women with chlamydial cervical infection showed no distinctive symptoms. Although associations were found between the presence of C. trachomatis and cervical discontinuity, purulent cervical exudate, and inflammatory changes in cervical cytology, these signs were not pathognomonic of chlamydial infection since they were also seen in some women infected with other microorganisms. There was a significant relationship between the presence of antibodies, detected by immunofluorescence, and the recovery of C. trachomatis in cell culture. There were no associations between ethnic group, history of sexually transmitted disease, method of contraception, phase in the menstrual cycle, or numbers of sexual contacts and infection with C. trachomatis.

Adolescent

Bacteriology of the urethra in normal men and men with nongonococcal urethritis.

Sixty-nine Caucasian males without a previous history of urethritis and who developed nongonococcal urethritis (NGU) and 39 similar men without urethritis (NU) were cultured from the urethra for Chlamydia trachomatis, Mycoplasma hominis, Ureaplasma urealyticum, aerobes, and anaerobes. C. trachomatis infection was proven by culture of serology in 26 (38%) of the NGU group and 1 (3%) of the NU group; the C. trachomatis-negative NGU group had significantly more U. urealyticum (81%) than the C. trachomatis-positive NGU group (42%) or the NU group (59%). Aerobes were isolated from significantly more NU men (91%) than from men with NGU (66%). The aerobic and anaerobic flora of the two NGU groups were similar. The NU group had significantly more aerobic lactobacilli. Haemophilus vaginalis, alpha-hemolytic streptococci (not Streptococcus faecalis), and anaerobes, predominantly Bacteroides species. This study has provided information about the prevalence and the variety of the aerobic and anaerobic microbiological flora of the anterior urethra of sexually active males. It does not implicate any bacteria other than C. trachomatis and U. urealyticum as potential causes of NGU.

Aerobiosis

CRISPR-Cas-based diagnostics for point-of-care detection of sexually transmitted infections: a laboratory development and evaluation study.

BACKGROUND: Timely, point-of-care diagnosis of sexually transmitted infections (STIs) is crucial for enabling prompt treatment and reducing transmission. We aimed to develop a portable, multiplexed, CRISPR-based assay panel for the detection of Neisseria gonorrhoeae (including the ciprofloxacin resistance marker gyrA S91F), Chlamydia trachomatis, Treponema pallidum, and herpes simplex virus (HSV). METHODS: In this laboratory development and evaluation study, we developed and optimised four multiplexed, CRISPR-based, diagnostic STI assays for point-of-care use. The complete assay panel comprised a CRISPR TP-HSV (cTP-HSV) panel for the detection of T pallidum and pan-HSV, with reflex testing to distinguish HSV-1 from HSV-2, and a CRISPR NG-CT (cNG-CT) panel for the detection of N gonorrhoeae and C trachomatis, with reflex testing to detect N gonorrhoeae using two additional genome regions and to identify the gyrA S91F mutation. Each pathogen was targeted at two independent genomic regions by isothermal amplification and CRISPR-Cas reaction using Cas12a and Cas13a, each with distinct fluorescent reporters. Analytical specificity and limits of detection (LODs) were determined, and a retrospective, masked concordance study was conducted on genomic DNA from 900 clinical samples (400 for cTP-HSV and reflex testing and 500 for cNG-CT and reflex testing), using quantitative PCR as the reference standard. The diagnostic accuracy of the test was assessed by analysis of receiver operating characteristic curves. FINDINGS: The overall sensitivity of the TP-HSV CRISPR assay was 82·5% (95% CI 74·0-88·7) for T pallidum and 94·4% (90·2-97·0) for pan-HSV; LODs were 6·2 copies per μL for T pallidum and 7·8 copies per μL for HSV. Reflex testing gave sensitivities of 97·0% (91·1-99·3) for HSV-1 and 96·0% (89·7-98·7) for HSV-2. The NG-CT CRISPR assay had an overall sensitivity of 80·0% (74·0-84·9) for N gonorrhoeae and 73·0% (65·5-79·3) for C trachomatis, with a LOD of 3·9 copies per μL for both pathogens. Reflex testing for the detection of the gyrA S91F mutation in N gonorrhoeae showed an overall sensitivity of 63·1% (55·1-70·4); however, this was dependent on sample type, with a sensitivity of 85·7% (46·7-99·5) in genital samples and 61·2% (52·8-68·9) in extragenital samples. For all pathogens, assay sensitivity was positively correlated with pathogen load. Area under the curve (AUC) values were 0·90 for T pallidum and 0·99 for pan-HSV in the TP-HSV assay, with values of 0·99 for HSV-1 and 0·97 for HSV-2 obtained in the reflex HSV-1-HSV-2 assay. For the cNG-CT assay, AUC values were 0·90 for N gonorrhoeae and 0·85 for C trachomatis, with a value of 0·72 obtained for gyrA S91F in the reflex cNG-gyrA assay. INTERPRETATION: Our multiplexed, CRISPR-based, point-of-care platform achieved performance consistent with WHO target product profiles for N gonorrhoeae and T pallidum. Proof-of-concept detection of the gyrA S91F resistance marker highlights its potential for resistance-guided therapy. Although optimisation is required before large-scale deployment, this suite offers a promising approach for rapid, decentralised, and resistance-informed STI diagnosis, particularly in resource-limited settings. FUNDING: Victorian Government Department of Health, Australian Government Department of Health, Disability and Ageing and Aged Care, and Australian Research Council.

Humans

Triple tetracycline (Deteclo) in the treatment of chlamydial infection of the female genital tract.

The efficacy of treating genital Chlamydia in women using Deteclo 300 mg twice daily for seven and 21 days has been studied. Forty-four patients were treated for seven days and 20 for 21 days. Seven days of treatment was as effective in eliminating Chlamydia trachomatis from the female genital tract as 21 days. The reproducibility of the technique of isolation of C. trachomatis was assessed by delaying treatment in 10 patients for up to 156 days and find C. trachomatis still present.

Chlamydia Infections

Chlamydia trachomatis infection of the Fallopian tubes. Histological findings in two patients.

In two patients with acute salpingitis, C. trachomatis was isolated from the cervix. In one of the patients, the organism was also recovered from the Fallopian tubes, and in the other, chlamydial inclusions were found in Giemsa-stained tubal epithelial cells. A significant change in micro-immunofluorescence antibodies to C. trachomatis occurred in both patients during the course of the disease. The Fallopian tubes of both patients were removed and studied by conventional histological techniques and, in the case of one of them, by transmission electron microscopy.

Adult

Light and electron microscopic study of Chlamydia trachomatis infection of the uterine cervix.

Cervical biopsy specimens from two patients with infections caused by Chlamydia trachomatis (TRIC agent) were studied by means of light and electron microscopy. Intraepithelial vesicles containing C. trachomatis were present in the cervical mucosa. These intramucosal vesicles were readily recognized in paraffin sections by means of light microscopy. The developmental stages of C. trachomatis, which have previously been seen in tissue cell culture, chicken embryo, and conjunctival infections, were also apparent in cervical infection.

Antibodies, Bacterial

Chlamydial pharyngitis?

Among 118 women who were sexual contacts of men with nongonococcal urethritis, the practice of fellatio correlated with symptoms of a sore throat. Oropharyngeal cultures for Chlamydia trachomatis were negative in all women, including 11 women who practiced fellatio and whose partners were known to have nongonococcal urethritis due to C. trachomatis. The study does not support a major role for C. trachomatis as a cause of sore throat in women who practice fellatio.

Chlamydia Infections

Isolation of Chlamydia trachomatis from Bartholin's ducts.

Exudate from Bartholin's ducts from 30 selected patients was investigated for Chlamydia trachomatis and Neisseria gonorrhoeae. N. gonorrhoeae was isolated from the duct exudate in 24 patients and C. trachomatis in nine. Concurrent infection of the ducts was present in seven (29.2%); the remaining two patients were sexual contacts of men with non-specific urethritis. The duct exudate was mucopus in seven patients, cloudy mucus in one, and clear mucus in the other. Although contamination of the vulva by C. trachomatis derived from cervical or urethral infection cannot be excluded, three cases are described which suggest that a true infection of the duct occurs and may persist after gonococcal infection has been cured.

Adolescent

Chlamydia trachomatis urethritis in men attending a venereal disease clinic: a culture and therapeutic study.

Urethral specimens from 459 male patients attending a venereal disease clinic were studied for presence of Chlamydia trachomatis and Neisseria gonorrhoeae. Of the 459 patients, 362 had symptoms suggestive of urethritis. In these 362, gonorrhoea was diagnosed in 78 (22%), while 88 (24%) harboured C. trachomatis; 15 harboured both organisms. Of those patients from whom C. trachomatis was isolated, 17% had no subjective symptoms of urethritis; the same percentage of symptomless carriers of gonococci was found. Watery discharge occurred more frequently in patients with chlamydial urethritis than in patients with gonorrhoea, while in the latter condition purulent discharge was more often found. Treatment with doxycycline for 9 days rendered 28 of 29 chlamydia-positive patients culture negative at control.

Bacteriological Techniques

Neonatal conjunctivitis caused by Neisseria gonorrhoeae and Chlamydia trachomatis.

In a selected group of 103 babies referred with neonatal conjunctivitis Neisseria gonorrhoeae was isolated from 11 and Chlamydia trachomatis from 33. Concurrent infection was present in three. On toddler sibling developed chlamydial conjunctivitis. After treatment C. trachomatis was re-isolated from six babies during the follow-up period. The discharge started one to three days after delivery in only three babies with gonococcal conjunctivitis and at five to eight days in eight babies. One baby was delivered by caesarean section. N. gonorrhoeae was isolated from four asymptomatic fathers, all of whom had urethritis. The mean onset of discharge in the 33 babies from whom Chlamydia was isolated was 7-1 days. One baby was delivered by caesarean section. Chlamydial conjunctivitis was associated with a high incidence of prematurity and of postpartum infection in the mother. Ten fathers of Chlamydia-positive babies were examined. C. tachomatis was isolated from four, all of whom were asymptomatic but had low-grade urethritis. These findings confirm the pathogenic role of C. trachomatis in the cervix and indicate the importance to the family of an adequate microbiological investigation of neonatal conjunctivitis.

Adult

Role of Chlamydia trachomatis in non-acute prostatitis.

The possible role of Chlamydia trachomatis in non-acute prostatitis was investigated by cultural and serological techniques in a study of 53 adult males. C. trachomatis was isolated from the urethra of only one of the 53 patients and from none of the 28 specimens of prostatic fluid from the same patients. By means of a modified microimmunofluorescent test, serum chlamydial IgG antibodies at a titre of 1/64 or greater, or IgM antibodies at a titre of 1/8 or greater, or both were detected in six of the patients, suggesting a recent or current chlamydial infection, while IgG or IgA antibodies at a titre of 1/8 or greater were detected in the specimens of prostatic fluid from two of the 28 men studied. In the seven patients with evidence of chlamydial infection, as well as in a further 13 of the 53 patients studied, the presenting symptoms suggested non-gonococcal urethritis (NGU) rather than prostatitis. Thus in this study C. trachomatis would appear to play a minor aetiological role, if any, in non-acute prostatitis.

Adult