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Capture-ELISA: a new assay for the detection of immunoglobulin M isotype antibodies using Chlamydia trachomatis antigen.

We present here a new method of IgM antibody-capture enzyme-linked immunosorbent assay (IgM-Capture-ELISA) for the diagnosis of recently acquired infections with Chlamydia trachomatis. For this analysis, plates were coated with goat IgG anti-human Fc mu. The capture of serum IgM antibodies was revealed indirectly by the sequential addition of biotinylated chlamydial proteins and peroxidase-conjugated streptavidin. In chlamydial extracts, cysteine-rich proteins are preferential antigenic targets for the humoral response. 3-(N-maleimidopropionyl)-biocytin (MPB), which binds biotinylated moieties to sulfhydryl groups, was used for the labeling procedure. The preservation of the antigenic specificity of labeled proteins was controlled by a blotting of these proteins, which were, respectively, probed either with specific IgM antibodies or with streptavidin. This analysis revealed that, after labeling, recognized epitopes are more particularly present on the major outer membrane protein (MOMP) of Chlamydia trachomatis. The validation of IgM-Capture-ELISA was assessed by using 170 selected sera from patients suspected of being infected by Chlamydia. Results were respectively compared to conventional indirect immunofluorescence assays (MIF-IgM assays) and to Western blotting. Sixteen sera were found to possess IgM antibodies against Chlamydia trachomatis with IgM-Capture-ELISA. Among these 16 sera, 14 and 15 were, respectively, positive with MIF-IgM assays and in Western blotting. Data obtained with IgM-Capture-ELISA reveal the absence of false-positive results in sera containing rheumatoid factor, which has been shown to interfere in the two other methods. IgM-Capture-ELISA value was then confirmed using sera from patients consulting for genital or pulmonary diseases, from patients with confirmed chlamydial infections, and from patients with other pathologies. IgM-Capture-ELISA appears as an alternative simple semi-quantitative assay for the detection of early chlamydial infection.

Animals↗

Chlamydia trachomatis: important relationships to race, contraception, lower genital tract infection, and Papanicolaou smear.

Chlamydia trachomatis is a common cause of sexually transmitted disease in adolescent girls. Of 366 adolescent patients screened, 15.3% were found to have chlamydial endocervical infections, with an infection rate of 23.3% in blacks, 14.3% in Hispanics, and 10.3% in whites (P = 0.01, excess for blacks). Of Chlamydia-positive patients, 63.6% had a diagnosis of lower genital tract infection, compared with 35.4% of Chlamydia-negative patients (P = 0.004). Oral contraceptive users had a higher prevalence of infection (23.8%) compared with those using a barrier method (16.2%) or with nonusers (9.3%) (P = 0.004). Inflammatory changes on Papanicolaou smears were associated with chlamydial infection (P = 0.0001). Other variables identified as risk factors for chlamydial infection included both a younger age at first intercourse (P = 0.02) and more years of sexual activity (P = 0.02). Chronologic, menarchal, and gynecologic age, biologic age of the cervix, the number of sexual partners in the last month and during a lifetime, and parity were not found to be associated with recovery of Chlamydia.

Chlamydia Infections↗

Isolation of Chlamydia trachomatis from the prostatic cells in patients affected by nonacute abacterial prostatitis.

To assess the presence of Chlamydia trachomatis in nonacute abacterial prostatitis 30 patients with urethral cultures positive for Chlamydia trachomatis underwent microbiological studies, including cultures of transrectal aspiration biopsies of the prostate. Chlamydia trachomatis was isolated from 10 of the prostatic specimens (33 per cent). In 3 cases a nonspecific cytopathogenic effect caused the destruction of the tissue cultures. Our findings demonstrate that Chlamydia trachomatis may cause ascending infections of the prostate and that this microorganism may have an etiological role in the pathogenesis of nonacute abacterial prostatitis.

Adult↗

[Prevalence of Chlamydia trachomatis infection, as evaluated by molecular biology methods].

BACKGROUND: To determine the prevalence of Chlamydia trachomatis infection in our area by molecular methods. METHODS: We describe the combined results of three studies carried out in the city of Barcelona including a total of 408 women considered to be at high risk for acquiring a sexually transmitted disease (STD). The first study was performed in 94 women attended at a public STD clinic located downtown, the second in 112 women attended at the Hospital Clinic and the third in 202 women attended at Hospital Vall d'Hebron (both third-level hospitals). In the first and third study endocervical exudate was tested with a PCR technique, and in the second study LCR was performed in 20-ml urine samples. RESULTS: Chlamydia trachomatis was detected in 1 woman in the first study, in no women in the second and in 3 women in the third. The prevalence of Chlamydia trachomatis infection detected in the three studies using molecular biology techniques was 1.06%, 0% and 1.48% respectively, giving an overall prevalence of 0.98%. CONCLUSION: The prevalence of Chlamydia trachomatis infection in our geographic area is surprisingly low.

Adult↗

High prevalence of Chlamydia trachomatis infections in adolescent females not having pelvic examinations: utility of PCR-based urine screening in urban adolescent clinic setting.

PURPOSE: To determine utility of polymerase chain reaction (PCR)-based urine screening for Chlamydia trachomatis in the care of adolescent females in an urban clinic. METHODS: Females > or = 15 years of age attending an adolescent clinic were approached consecutively. Each enrollee was interviewed to determine the primary reason(s) for the clinic visit and was queried about genitourinary symptoms. Nonsterile voided urine specimens were tested for C. trachomatis using PCR-based analysis. Endocervical C. trachomatis cultures were obtained from the subjects who had a pelvic examination. Main outcome measures were chlamydia infection rates in clinic attendees whether a pelvic examination was performed or not. RESULTS: A total of 315 (99.4%) of 317 patients approached agreed to participate. Overall, 47 (14.9%) patients had positive urine PCR tests. The chlamydia infection rate detected by urine PCR was 22.1% (19 of 86) among those who had pelvic examinations performed and 12.2% (28 of 229) among those who did not (p = .03; odds ratio 2.04; 95% confidence interval 1.02, 4.06). Sixty percent (28 of 47) of chlamydia infections identified during the study period were identified by the urine screening test. CONCLUSION: Urine screening was accepted by vast majority of female adolescents attending the clinic irrespective of reason for the clinic visit, and was highly effective in identifying unsuspected C. trachomatis infections, particularly among girls attending the clinic for reasons unrelated to reproductive health care and as an interim screening tool for adolescent family-planning clients.

Adolescent↗

Sexual behavior and the prevalence of Chlamydia trachomatis infection in asymptomatic students in Germany and Spain.

This study aimed to assess patterns of sexual activity, preventive behaviors, contraceptive use, and the prevalence of chlamydia infection in student populations from two countries. 188 asymptomatic students of the University of Bielefeld, Germany and 590 students of the Navarra Public University, Spain were surveyed using a standardized self-administered questionnaire. Urine samples were analyzed by ligase chain reaction technique for chlamydia infection. Compared to German students, Spanish students were less likely to have more than one sex partner within the last 12 months (OR: 0.62; CI: 0.46-0.83), to have sexual intercourse more than one time per week (OR: 0.70; CI: 0.52-0.93) and to use oral contraceptives (OR: 0.16; CI: 0.10-0.26). They reported a higher use of condoms (OR: 2.93; CI: 2.01-4.27) and were more likely to always use condoms with a new sex partner (OR: 2.47; CI: 1.72-3.53). The prevalence of chlamydia infection was considerably higher in German students (5.8% [corrected] in females; 2.2% in males) than in Spanish students, where no case was found. The higher frequency of sexual activity, a higher engagement in risk-taking sexual behavior and the lower use of barrier contraceptives may contribute to the higher prevalence of chlamydia infection in German students.

Adult↗

Acute and chronic urinary tract infections caused by Chlamydia trachomatis.

Chlamydia trachomatis is one of the causes of acute and chronic urinary tract infections and acute or silent salpingitis. Chronic or recurrent female urinary or genital tract infections with Chlamydia trachomatis have been recognised as a significant factor in the development of acute or chronic renal interstitial inflammation or increased risk of ectopic pregnancy. In most cases Chlamydia trachomatis is sexually transmitted. Moreover, it is one of the most common sexually transmitted pathogens. The current estimate is that in the United States there occur 4.5 million new infections each year. We describe 3 cases of recurrent urinary tract infections due to Chlamydia trachomatis.

Acute Disease↗

Chlamydia and apoptosis: life and death decisions of an intracellular pathogen.

The chlamydiae are important obligate intracellular prokaryotic pathogens that, each year, are responsible for millions of human infections involving the eye, genital tract, respiratory tract, vasculature and joints. The chlamydiae grow in cytoplasmic vesicles in susceptible host cells, which include the mucosal epithelium, vascular endothelium, smooth muscle cells, circulating monocytes and recruited or tissue-specific macrophages. One important pathogenic strategy that chlamydiae have evolved to promote their survival is the modulation of programmed cell death pathways in infected host cells. The chlamydiae can elicit the induction of host cell death, or apoptosis, under some circumstances and actively inhibit apoptosis under others. This subtle pathogenic mechanism highlights the manner in which these highly successful pathogens take control of infected cells to promote their own survival - even under the most adverse circumstances.

Apoptosis↗

Chlamydia trachomatis utilizes the host cell microtubule network during early events of infection.

The host cell cytoskeleton is known to play a vital role in the life cycles of several pathogenic intracellular microorganisms by providing the basis for a successful invasion and by promoting movement of the pathogen once inside the host cell cytoplasm. McCoy cells infected with Chlamydia trachomatis serovars E or L2 revealed, by indirect immunofluorescence microscopy, collocation of microtubules and Chlamydia-containing vesicles during the process of migration from the host cell surface to a perinuclear location. The vast majority of microtubule-associated Chlamydia vesicles also collocated with tyrosine-phosphorylated McCoy cell proteins. After migration, the Chlamydia-containing vesicles were positioned exactly at the centre of the microtubule network, indicating a microtubule-dependent mode of chlamydial redistribution. Inhibition of host cell dynein, a microtubule-dependent motor protein known to be involved in directed vesicle transport along microtubules, was observed to have a pronounced effect on C. trachomatis infectivity. Furthermore, dynein was found to collocate with perinuclear aggregates of C. trachomatis E and L2 but not C. pneumoniae VR-1310, indicating a marked difference in the cytoskeletal requirements for C. trachomatis and C. pneumoniae during early infection events. In support of this view, C. pneumoniae VR-1310 was shown to induce much less tyrosine phosphorylation of HeLa cell proteins during uptake than that seen for C. trachomatis.

Animals↗

Chlamydia pneumoniae and multiple infections in the aorta contribute to atherosclerosis.

Our previous study on herpesvirus infection including Herpes simplex virus type 1 (HSV-1), Epstein-Barr virus (EBV), cytomegalovirus (CMV) and atherosclerosis revealed that the prevalence of herpesvirus is higher in atherosclerotic aorta than in non-atherosclerotic aorta. Infections with two or three forms of the virus have been found only in atherosclerotic aorta. In our current study, we examined both Chlamydia pneumoniae and Chlamydia trachomatis in herpesvirus-infected aortic tissues, by means of immunohistochemistry, polymerase chain reaction, Southern hybridization, in situ hybridization, electron microscopy and electron-microscopic immunohistochemistry. In particular, the bacteria were found in atherosclerotic lesions. In atherosclerotic aorta, 40% of tissues examined were positive for C. pneumoniae in contrast to absence of this bacteria in non-atherosclerotic aorta. Elementary bodies of C. pneumoniae were found in macrophage-like cells in the intima of atherosclerotic aorta by electron microscopy. Chlamydia trachomatis was not found in both atherosclerotic and non-atherosclerotic aorta. Our findings suggest that multiple infections in aortic tissue contribute to the development of atherosclerosis. Furthermore, the absence of C. pneumoniae compared to herpesviruses in normal arterial tissue suggests that C. pneumoniae is specific for atherosclerotic lesions. In contrast to 'abortive infection' of viruses in arteries, C. pneumoniae infection was demonstrated in macrophages by electron microscopy and electron-microscopic immunohistochemistry in atherosclerotic lesion. Chlamydia pneumoniae may be the most important pathogen related to the development of atherosclerosis.

Adolescent↗

[Chlamydia infections in gynecology and obstetrics].

Urogenital infections with Chlamydia trachomatis belong to the most prevalent sexual transmitted diseases worldwide. In women, Chlamydia trachomatis can cause an infection of the cervix, the urethra, the endometrium and the Fallopian tubes. As potential sequelae, tubal infertility and ectopic pregnancies are associated with urogenital infections caused by this pathogen. An infection of the cervix in pregnancy can lead to neonatal and maternal complications. Therefore, by April 1st, 1995 a cervical swab for Chlamydia trachomatis has been included in the German antenatal care guide lines. The significance of these infections in health care is in contrast to its often hardly symptomatic or asymptomatic course. New diagnostic methods facilitate the detection of the pathogen and allow a specific antibiotic therapy. The aim of this review is a presentation of the clinical significance, diagnosis and therapy of urogenital infections with Chlamydia trachomatis in obstetrics and gynaecology to contribute to the prevention of potentially severe sequelae.

Anti-Bacterial Agents↗

[Is there a correlation between tubal occlusions in chronic salpingitis and urogenital chlamydia infections?].

A prospective study was performed to analyse the relationship between urogenital infections caused by Chlamydia trachomatis and occlusions of the fallopian tubes with histologically confirmed chronic salpingitis and salpingitis isthmica nodosa. 110 infertile patients were tested for C. trachomatis infection. 23 patients with tubal occlusions and histologically confirmed chronic salpingitis (group 1) and eight patients with salpingitis isthmica nodosa (group 2) were compared to 13 patients with tubal occlusions after tuboligation (group 3), and to 66 patients with patent fallopian tubes as demonstrated by laparoscopy or hysterosalpingography (group 4). The prevalence of infections of the endocervix or urethra and the presence of Chlamydia in urine was low in all four groups. However, in groups 1 and 2, the median Chlamydia IgG and IgA serum antibody titres were significantly higher (p < or = 0.0002) than in groups 3 and 4. This result illustrates the association between urogenital infections with Chlamydia and tubal occlusions with histologically documented chronic salpingitis and salpingitis isthmica nodosa.

Adult↗

[Chlamydia and mycoplasma infections in men of couples with involuntary sterility].

There is conflicting evidence on the influence of infections with Chlamydia trachomatis and Mycoplasma hominis on male and female infertility. We studied the prevalence of Chlamydia and Mycoplasma in male partners of 165 infertile couples. 25% of couples with tubal and/or andrological sources of infertility showed positive cultures for Chlamydia and/or Mycoplasma compared with 10% of couples with other causes of infertility. Our data suggest, that screening for Chlamydia and Mycoplasma in infertility patients may be of assistance.

Adult↗

[Significance of Chlamydia trachomatis for female fertility with special reference to prevalence in the endometrium].

144 patients with longstanding infertility (mean duration of infertility 4.9 years) were screened for endometrial infection with Chlamydia trachomatis by means of endometrial biopsy and McCoy cell-culture. The same day blood was taken for detection of Chlamydia IgG antibodies in serum samples. During a 1 year's period, all new patients of a large infertility clinic without complaints or clinical symptoms of infection of the lower genital tract were consecutively submitted to the study. Elevated titers of IgG antibodies in serum samples (Chlam. AB greater than or equal to 1/256) revealed previous chlamydial infection of 26% of patients, but Chlamydia trachomatis was detected in endometrial biopsies in only 5/144 (3.5%). There was a strong correlation of previous chlamydial infection and reduced tubal patency (p 0.001) and a lower pregnancy rate after 6 months. The results suggested that infections with Chlamydia trachomatis, influenced by demographic and socio-economic aspects, are of minor importance as an actual infection in asymptomatic patients with longstanding infertility. Previous chlamydial infections, however, are of great importance because of their negative influence on tubal function.

Adult↗

[Clinical and serological characterization of Chlamydia-induced arthritis].

Chlamydia-induced arthritis was diagnosed in 32 patients (18 men and 14 women; mean age 33 [17-54] years). 13 were HLA-B27 positive. In 19 patients the arthritis was largely confined to one joint. Enthesopathies occurred in 14 patients, and in 13 sacroiliitis was diagnosed radiologically. The most frequent extra-articular changes affected the urogenital system (26 cases). The full Reiter syndrome was present in seven patients (four men, three women). All patients had serum antibodies against Chlamydia. In 16 patients Chlamydia was diagnosed as the causative organism by urogenital swab. Chlamydia-induced arthritis differs from other arthritides in the high proportion of HLA-B27 negative patients.

Adolescent↗

The tip of the iceberg: opportunistic screening for Chlamydia trachomatis in asymptomatic patients attending a young people's health clinic reveals a high prevalence--a pilot study.

OBJECTIVES: We implemented an opportunistic screening programme for Chlamydia trachomatis amongst patients presenting to a young peoples' health service in the city of Geelong, Australia, to define the prevalence of infection and to identify specific risk factors. METHODS: Over a 7-month period sexually active patients attending the young peoples' clinic were offered C. trachomatis screening by nucleic acid amplification test. There was 100% acceptance rate among those offered the test. Patient demographics, reason for presentation at the clinic and reported symptoms were documented by the clinicians and correlated with laboratory findings. RESULTS: 163 patients between the ages of 12-25 were tested, nine males and 154 females. The prevalence of chlamydia infection was 5.8% and was highest (16.0%) among patients presenting for the morning after pill. Inhibition of the nucleic acid amplification test occurred in 11.0% of urine samples. All patients with inhibited tests were asked to provide a repeat sample for retesting, but only 50% complied with this request. The majority of repeat samples (88.9%) had no inhibitors present and yielded a negative result. There was no correlation between symptoms and a positive chlamydia result. CONCLUSIONS: Chlamydia infection is common in young people engaging in unsafe sexual practice and cannot be predicted by the presence of symptoms. The high prevalence of infection in Geelong would make screening cost effective in this age group. Ongoing population screening of sexually active young people should be encouraged in community health centres. Inhibition of the nucleic acid amplification test was common but repeat testing of urine a few days later usually gave satisfactory results.

Adolescent↗

Genital Chlamydia trachomatis infection in Australia: epidemiology and clinical implications.

Over the last decade, notification rates for genital Chlamydia trachomatis infection in Australia have been rising progressively. While chlamydia is common and possibly increasing in the general population, heterosexual adolescents, indigenous Australians in remote settings, and homosexually active men are at particular risk of infection. Few studies are available on the extent of morbidity from chlamydia-associated diseases. Australia urgently needs a national strategy to control chlamydia, with widespread, selective screening as a key component. As general practitioners have an important role to play, we proffer guidelines for selective testing in primary care.

Australia↗

The quantity of nitric oxide released by macrophages regulates Chlamydia-induced disease.

Intracellular bacteria of the genus Chlamydia cause numerous typically chronic diseases, frequently with debilitating sequelae. Genetic determinants of disease susceptibility after infection with Chlamydia bacteria are unknown. C57BL/6 mice develop severe pneumonia and poor immunity against Chlamydia after moderate respiratory infection whereas BALB/c mice are protected from disease and develop vigorous Th1 immunity. Here we show that infected C57BL/6 macrophages release more NO synthesized by NO synthase 2 (NOS2) than BALB/c macrophages and have lower mRNA concentrations of arginase II, a competitor of NOS2 for the common substrate, l-arginine. Reduction, but not elimination, of NO production by incomplete inhibition of NOS2 abolishes susceptibility of C57BL/6 mice to Chlamydia-induced disease. Thus, the quantity of NO released by infected macrophages is the effector mechanism that regulates between pathogenic and protective responses to chlamydial infection, and genes controlling NO production determine susceptibility to chlamydial disease.

Animals↗