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Detection of Chlamydia trachomatis in fallopian tube tissue in women with postinfectious tubal infertility.

OBJECTIVE: Biopsy tissues from women with postinfectious tubal infertility were studied for the presence of Chlamydia trachomatis. STUDY DESIGN: Tubal biopsy specimens from 25 women with postinfectious tubal infertility undergoing laparoscopy for repair of fallopian tubes were evaluated by culture, in situ hybridization. Immunocytochemistry, and transmission electron microscopy for the presence of Chlamydia trachomatis. Serum was also tested for Chlamydia trachomatis antibodies. RESULTS: Chlamydia trachomatis was detected in postinfectious tubal biopsy specimens in three of 25 patients by culture, 12 of 24 by in situ hybridization, 15 of 22 by immunoperoxidase stain, and two of 10 by transmission electron microscopy. Serum antibody against Chlamydia trachomatis was detected in 15 of 21 patients. CONCLUSION: Chlamydia trachomatis deoxyribonucleic acid or antigens were detected at a high percentage (19/24 women) in the biopsy tissues of the fimbrial and peritubal adhesions by in situ hybridization or immunoperoxidase stain, suggesting a persistent infection in these women even after antibiotic treatment.

Adult↗

Chlamydia trachomatis and Ureaplasma urealyticum in men attending a sexually transmitted diseases clinic.

Urethral specimens from 480 heterosexual patients were examined for Chlamydia trachomatis. Chlamydia were isolated from 32.7% of men with non-specific urethritis (NSU), from 16.1% of men with gonorrhoea, and from 4.1% of men without urethritis. Chlamydial isolation was not related to duration of symptoms, presence of discharge, or past history of attendance at the clinic. Urine from 176 heterosexual patients was examined for Ureaplasma urealyticum. Ureaplasmas were present in 53.8% of men with NSU, in 28% of men with gonorrhoea, and in 32.9% of men with no urethritis. Detection rates for ureaplasmas in patients with chlamydia-negative and chlamydia-positive NSU were similar, but ureaplasmas were present in significantly greater numbers in patients with chlamydia-negative NSU than in those with chlamydia-positive NSU.

Animals↗

Detection of seroconversion and persistence of Chlamydia trachomatis antibodies in five different serological tests.

Microimmunofluorescence (MIF), a Chlamydia trachomatis species-specific enzyme immunoassay incorporating lipopolysaccharide-extracted Chlamydia trachomatis L2 elementary bodies, two different synthetic peptide-based species-specific tests, and a recombinant lipopolysaccharide genus-specific test were performed on multiple follow-up sera (n = 104 total) from 16 women with Chlamydia trachomatis-positive cervical swabs. These women included five with IgG seroconversions, five with Chlamydia trachomatis reinfections after initial therapy, and six with serologic follow-up of more than 6 years after antibiotic therapy. Of all the tests employed in this study, MIF IgG reverted earliest to negative titers, while MIF IgA was the least sensitive. The lipopolysaccharide-extracted elementary body enzyme immunoassay exhibited the closest correlation with the MIF test. The highest test sensitivity was observed in one of the synthetic peptide-based tests, which detected earliest seroconversions and longest IgG persistence. The other synthetic peptide-based test gave false-negative results in 2 of 16 women and did not detect seroconversion earlier than the MIF test. Seroconversion and persistence of genus-specific IgG--cross-reactivity with Chlamydia pneumoniae--against lipopolysaccharide were similar to species-specific IgG. A significant serologic response to reinfection was observed only in women with signs of pelvic inflammatory disease. Species-specific tests of high sensitivity and reproducibility are best suited for gynecological diagnostic purposes.

Adult↗

HLA-DP restricted Chlamydia trachomatis specific synovial fluid T cell clones in Chlamydia induced Reiter's disease.

Synovial fluid (SF) mononuclear cells from a patient with Chlamydia trachomatis induced acute Reiter's disease were directly by limiting dilution in a representative protocol using phytohemagglutinin in the cloning medium. Out of 76 alpha beta-TCR+ CD4+ T lymphocyte clones, 7 were shown to specifically recognize C. trachomatis in a proliferation assay. The antigen recognition of these clones was HLA-DP restricted. Unexpectedly, 2 HLA-DR restricted clones showed a proliferative response to Yersinia enterocolitica O3, though the patient had no history of yersinia infection. The high frequency of SF derived T cells with specificity for species-specific chlamydial antigens and the limited diversity of HLA class II restriction of these clones may indicate an oligoclonal synovial T cell response to persistent intraarticular chlamydia.

Adult↗

Detection of Chlamydia trachomatis by the polymerase chain reaction in the cervices of women with acute salpingitis.

OBJECTIVE: Our objective was to determine whether an increased prevalence of Chlamydia trachomatis could be detected by the polymerase chain reaction as opposed to culture in the cervices of women with acute salpingitis. STUDY DESIGN: Endocervical samples from 15 women with laparoscopy-verified acute salpingitis and 20 women seeking medical help for conditions other than pelvic pain were tested for Chlamydia trachomatis with the polymerase chain reaction. The oligonucleotide primer pairs used were specific for a 144 bp region of the major outer membrane protein that contained a single EcoRI endonuclease cleavage site. The detection of a 144 bp band that was cleaved by EcoRI to a 103 bp band denoted a Chlamydia trachomatis-positive sample. Cervical samples were cultured for Chlamydia trachomatis with the use of McCoy cells. The lymphocyte proliferative response to Chlamydia trachomatis elementary bodies was also determined. RESULTS: Nine of the 15 women (60%) with salpingitis had positive results when tested with the polymerase chain reaction for cervical Chlamydia trachomatis. Only two of these women (13%), both of whom had positive results when tested with the polymerase chain reaction, had cultures that were positive for Chlamydia trachomatis (p < 0.02). Among the 20 other women, only two patients with cervicitis had positive cultures for Chlamydia. Those women plus two women with unexplained recurrent abortions had positive polymerase chain reaction test results for Chlamydia trachomatis. A lymphocyte proliferative response to Chlamydia trachomatis was detected in five of eight women with salpingitis, as well as three of the other four patients, all of whom had positive polymerase chain reaction test results; lymphocytes from the remaining women were unresponsive. Follow-up cervical samples were obtained 4 to 6 months after treatment from six of the patients with salpingitis who had positive polymerase chain reaction test results; at that time five had negative polymerase chain reaction test results for Chlamydia trachomatis. CONCLUSION: The polymerase chain reaction appeared to be more sensitive and more rapid than culture in detecting Chlamydia trachomatis in the cervices of women with acute salpingitis. This assay may be of value for the early diagnosis of chlamydial infections.

Acute Disease↗

Efficacy of single-dose azithromycin versus doxycycline in the treatment of cervical infections caused by Chlamydia trachomatis.

The efficacy of single-dose azithromycin therapy in the treatment of cervical Chlamydia trachomatis infections was compared to that of a standard seven-day course of treatment with doxycycline. Cervical samples from 60 patients reacted positively in an enzyme immunoassay for detection of Chlamydia trachomatis. In 31 patients Chlamydia trachomatis was isolated from the sample taken before treatment. Fourteen of the 31 patients were treated with doxycycline and 17 with azithromycin. All cultures of samples taken one and four weeks after the start of therapy were negative. All 31 isolates showed a similar pattern of MICs for the seven antibiotics tested, including azithromycin and doxycycline. No differences were observed between isolates of different serovars. In samples from four patients chlamydial DNA could be detected by PCR one week after the start of the therapy and in two patients also after four weeks. No difference in microbiological parameters could be observed between the two treatment groups. It is concluded that single-dose azithromycin is as effective as a seven-day course of doxycycline in the therapy of cervical Chlamydia trachomatis infections.

Azithromycin↗

The vaginal introitus: a novel site for Chlamydia trachomatis testing in women.

OBJECTIVE: We evaluated the vaginal introitus as a noninvasive sampling site for testing for Chlamydia trachomatis. STUDY DESIGN: Swabs from the vaginal introitus were obtained from 300 women attending a sexually transmitted diseases clinic and tested for the presence of Chlamydia trachomatis by polymerase chain reaction. Additionally, 200 of these women self-collected an additional introitus swab and submitted a urine sample for polymerase chain reaction testing. These samples were compared with polymerase chain reaction, culture, and enzyme immunoassay for Chlamydia trachomatis from endocervical samples and polymerase chain reaction and culture on urethral swabs. Patients were determined to be infected with Chlamydia trachomatis by a positive culture result from any site or a confirmed positive result by polymerase chain reaction with an alternate primer. RESULTS: The sensitivity of vaginal introitus swabs obtained by health care providers for the detection of urogenital Chlamydia trachomatis was 92% (95% confidence interval 83 to 100), greater than polymerase chain reaction, culture, or enzyme immunoassay of the cervix or urethra. The sensitivity by polymerase chain reaction of patient self-collected swabs was 81%. Sampling of the vaginal introitus, by both health care workers and the patient herself, performed as well as commonly used diagnostic tests that require vaginal speculum examination. The sensitivity of polymerase chain reaction testing of urine samples was 73%. CONCLUSION: The vaginal introitus represents a highly effective noninvasive specimen collection site for Chlamydia trachomatis testing by polymerase chain reaction. Self-collection of introitus specimens may revolutionize sexually transmitted disease testing by eliminating the need for a speculum examination by skilled health care personnel.

Chlamydia Infections↗

Unusual manifestations of Chlamydia trachomatis infections.

Systemic infections with Chlamydia trachomatis are known to occur with the agents of lymphogranuloma venereum but are not generally recognized to occur with the trachoma and inclusion conjunctivitis (TRIC) agents, i.e., immunotypes A-K. The clinical spectrum of TRIC agent infections has expanded, however, and now includes deep-seated genital infections such as epididymitis and salpingitis, as well as infections in neonates. Endocarditis, pneumonia in adults, otitis media, choroiditis and erythema nodosum are unusual manifestations of C. trachomatis infections that may be seen. Meningoencephalitis, chronic palmoplantar pustulosis, and pituriasis rosea also might be associated with C. trachomatis infection. Finally, lymphogranuloma venereum may have systemic manifestations, and Chlamydia psittaci infections may be characterized by extrapulmonary involvement.

Adult↗

Prevalence of urogenital Chlamydia trachomatis increases significantly with level of urbanisation and suggests targeted screening approaches: results from the first national population based study in the Netherlands.

OBJECTIVES: Chlamydia trachomatis (Chlamydia) is the most prevalent sexually transmitted bacterial infection and can cause considerable reproductive morbidity in women. Chlamydia screening programmes have been considered but policy recommendations are hampered by the lack of population based data. This paper describes the prevalence of Chlamydia in 15-29 year old women and men in rural and urban areas, as determined through systematic population based screening organised by the Municipal Public Health Services (MHS), and discusses the implications of this screening strategy for routine implementation. METHODS: Stratified national probability survey according to "area address density" (AAD). 21 000 randomly selected women and men in four regions, aged 15-29 years received a home sampling kit. Urine samples were returned by mail and tested by polymerase chain reaction (PCR). Treatment was via the general practitioner, STI clinic, or MHS clinic. RESULTS: 41% (8383) responded by sending in urine and questionnaire. 11% (2227) returned a refusal card. Non-responders included both higher and lower risk categories. Chlamydia prevalence was significantly lower in rural areas (0.6%, 95% CI 0.1 to 1.1) compared with very highly urbanised areas (3.2%, 95% CI 2.4 to 4.0). Overall prevalence was 2.0% (95% CI 1.7 to 2.3): 2.5% (95% CI 2.0 to 3.0%) in women and 1.5% (95% CI 1.1 to 1.8) in men. Of all cases 91% were treated. Infection was associated with degree of urbanisation, ethnicity, number of sex partners, and symptoms. CONCLUSION: This large, population based study found very low prevalence in rural populations, suggesting that nationwide systematic screening is not indicated in the Netherlands and that targeted approaches are a better option. Further analysis of risk profiles will contribute to determine how selective screening can be done.

Adolescent↗

Prevalence of Chlamydia trachomatis infection in a population of asymptomatic women in a screening program for cervical cancer.

The prevalence of Chlamydia trachomatis infection in a population of women with no symptoms of sexually transmitted disease was investigated. These women, aged 35-55 years, participated in a screening program for cervical cancer. With the use of a direct immunofluorescence method, 109 out of 2,470 smears tested were positive for Chlamydia trachomatis, indicating an overall prevalence of 4.4%. No changes in prevalence were found when five-year cohorts of this group were analyzed, indicating that age-dependent changes or epidemiological factors do not result in a different (decreased) prevalence over the ages 35 to 55 years. The prevalence of Trichomonas vaginalis and fungi, as detected by cytological screening, was lower than that observed for Chlamydia trachomatis: 3.1 and 2.1%, respectively. Of the 109 smears positive for Chlamydia trachomatis, 90 showed cervical cells with reactive changes (out of 1,490 smears with PAP II), whereas no cytological changes were found in 15 cases (out of 884 smears with PAP I). Changes suggestive of mild or moderate dysplasia were found in only four cases (out of 93 smears with PAP III). The results indicate that Chlamydia trachomatis is associated with reactive changes of endocervical cells and raise serious questions about whether prevention of possible secondary effects such as infertility and pelvic inflammatory disease can be achieved by a combined screening program for cervical cancer and Chlamydia trachomatis.

Adult↗

DNA amplification assays: a new standard for diagnosis of Chlamydia trachomatis infections.

Infections caused by Chlamydia trachomatis are among the most common bacterial sexually transmitted infections in the world with an estimated 50 million new cases occurring each year. Since the prevalence of chlamydial infections range from 3%-5% among asymptomatic men and women to 20%-25% among adolescents attending sexually transmitted disease (STD) clinics, widespread screening for this infection has been recommended. With the advent of DNA amplification technology, several assays have become commercially available which have high sensitivity and specificity, and can utilise convenient, non-invasive specimens such as urine. Using the endogenous plasmid of C trachomatis as target DNA, polymerase chain reaction (PCR) and ligase chain reaction (LCR) have demonstrated sensitivities of 90% to 96%, with specificities of 98% to 100%. In contrast, culture, the previous gold standard for C trachomatis detection, had sensitivities of 50% to 70% for male urethral specimens and 60% to 80% for female endocervical specimens when compared to confirmed PCR and LCR. DNA sequencing and restriction fragment length polymorphisms (RFLP) of amplified target DNA from the major outer membrane protein gene (OMPI) of C trachomatis has been performed to verify specificity and to study the molecular epidemiology of C trachomatis. Nucleic acid amplification assays have proven to be effective in the detection of chlamydia in other clinical specimens including ocular specimens from patients with trachoma, epididymitis, salpingitis, and reactive arthritis. Urine-based screening of both men and women by LCR or PCR provides a unique sensitive method for widespread population-based screening for C trachomatis, a necessary tenet of chlamydia control programmes.

Chlamydia Infections↗

[Chlamydia trachomatis infections of the cervix and postoperative course].

In the paper authors consider the problem if Chlamydia trachomatis infection of urogenital tract exerts an influence on postoperative course in women who were operated in the gynaecology department. 186 women have been examined in this study. Using the criteria for selective screening for Chlamydia trachomatis infection proposed by Handsfield, 29 women with higher risk for chlamydial infection were seperated. The personal details have been obtained by standardized questionnaire. Using the immunoenzymatic test "Chlamydiazyme" the incidence of Chlamydia trachomatis infection of the cervical canal was studied in 29 women from groups of higher risk for this infection. No Chlamydia trachomatis antigens were found in this group. These results don't answer the question if Chlamydia trachomatis infection of urogenital tract exerts an influence on postoperative course in women who were operated in gynaecology department, because it is to small trial.

Chlamydia Infections↗

Diagnostic methods for Chlamydia trachomatis disease in neonates.

Chlamydia trachomatis is an obligate intracellular parasite responsible for many clinical syndromes, including neonatal conjunctivitis and pneumonia. The gold standard of diagnosis has been isolation in cell culture. However, this requires days of processing. Several rapid diagnostic tests are available. Giemsa staining of conjunctival smears, enzyme immunoassay, and the fluorescein-conjugated monoclonal antibody test. Both the EIA and the FA tests show promise as ideal rapid diagnostic tests. Treatment of chlamydial conjunctivitis must focus upon the eradication of nasopharyngeal carriage as well as cure of ophthalmic symptoms. The need for nasopharyngeal eradication is underscored by the fact that it is the source for chlamydial pneumonia as well as for conjunctival re-infection. Clinical studies have shown that oral erythromycin estolate or ethylsuccinate suspension 50 mg/kg/day twice-daily or four times a day for 14 to 21 days are the therapeutic regimens of choice. Neonatal ocular prophylaxis is currently under study. One per cent silver nitrate does not prevent chlamydial conjunctivitis but preliminary studies do show favorable results with topical erythromycin. Nevertheless, neither 1 per cent silver nitrate nor topical erythromycin eradicate nasopharyngeal carriage, elimination of which is necessary for the prevention of neonatal chlamydial pneumonia.

Conjunctivitis, Inclusion↗

Characteristics of the m2000 automated sample preparation and multiplex real-time PCR system for detection of Chlamydia trachomatis and Neisseria gonorrhoeae.

We evaluated a new real-time PCR-based prototype assay for the detection of Chlamydia trachomatis and Neisseria gonorrhoeae developed by Abbott Molecular Inc. This assay is designed to be performed on an Abbott m2000 real-time instrument system, which consists of an m2000sp instrument for sample preparation and an m2000rt instrument for real-time PCR amplification and detection. The limit of detection of this prototype assay was determined to be 20 copies of target DNA for both C. trachomatis and N. gonorrhoeae, using serially diluted linearized plasmids. No cross-reactivity could be detected when 55 nongonococcal Neisseria isolates and 3 non-C. trachomatis Chlamydia isolates were tested at 1 million genome equivalents per reaction. Concordance with the Roche Amplicor, BDProbeTec ET, and Gen-Probe APTIMA Combo 2 tests was assessed using unlinked/deidentified surplus clinical specimens previously analyzed with these tests. For C. trachomatis, concordance for positive results ranged from 93.7% to 100%, while concordance for negative results ranged from 98.2% to 100%. For N. gonorrhoeae, concordance for positive and negative results ranged from 91.4% to 100% and 99.3% to 100%, respectively. A workflow analysis of the prototype assay was conducted to obtain information on throughput under laboratory conditions. At 48 samples/run, the time to first result for both C. trachomatis and N. gonorrhoeae was 4.5 h. A total of 135 patient specimens could be analyzed in 8.9 h, with 75 min of hands-on time. This study demonstrated the technical and clinical feasibility of the new Abbott real-time PCR C. trachomatis/N. gonorrhoeae assay.

Automation↗

The role of calf serum in the growth of Chlamydia trachomatis in McCoy cell cultures.

Chlamydia trachomatis is normally grown in McCoy monolayer coverslip cultures using partially defined media containing foetal calf serum at concentrations up to 10% (v/v). Omission of the serum decreased the number of inclusions produced by infecting the McCoy cells with a standard inoculum of a genital strain of C. trachomatis. Substitution of the foetal calf serum with a macromolecular fraction from the serum or with a mixture of sodium oleate, bovine serum albumin fraction V and fetuin maintained the inclusion count, but substitution with serum filtrate, containing the amino acids and other low molecular weight components, decreased the inclusion count of C. trachomatis. The role of calf serum and the need for a fully defined medium excluding serum for studying the growth of C. trachomatis in tissue culture are discussed.

Animals↗

Intrauterine Chlamydia trachomatis infection in a premature infant.

Intrauterine Chlamydia trachomatis infection was strongly suspected in a premature infant born in the 32nd week of gestation. The membranes were artificially ruptured at the time of delivery. This infant showed a high titer of specific IgM antibody to Chlamydia trachomatis at one hour after birth. He showed mild respiratory distress and was treated with oral erythromycin for three weeks. He was discharged home at the age of 46 days.

Chlamydia Infections↗

A microbiological study of conjunctivitis with emphasis on Chlamydia trachomatis, in northern Norway.

To determine the microbiological agents in conjunctivitis in children and young adults, physicians outside hospitals were asked to obtain samples from the conjunctiva in patients presenting with conjunctivitis. Specimens from 194 patients and 177 healthy controls were cultivated for Chlamydia trachomatis. In 12 cases Chlamydia trachomatis was isolated (i.e. 4 neonates, 1 three-year-old child and 7 adults, aged 17 to 39 years), but in none of the controls. Of the specimens from patients 168 and all of the healthy controls were cultivated for both bacteria (including Chlamydia trachomatis) and viruses. The main micro-organisms, regarded as infectious, were Haemophilus influenzae (20), Streptococcus pneumoniae (18), Staphylococcus aureus (14) and Chlamydia trachomatis (9). Haemophilus influenzae (non-typable strains) were isolated more frequently in the age group below 5 years of age than in the age group 5-50 years. Herpes simplex virus (type II) was isolated in one neonate. Chlamydia trachomatis is among the most important infectious agents in conjunctivitis treated outside hospitals. As chlamydial infections need special attention regarding treatment and follow-up, physicians should be encouraged to obtain specimens for microbiological examination, including chlamydia, from the population at risk.

Adolescent↗