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The prevalence of Chlamydia trachomatis infection in the genitourinary tract is increasing, with an annual rise of 9 million cases. Individuals afflicted with these infections are at a heightened risk of developing adult inclusive conjunctivitis (AIC), which is commonly recognized as the ocular manifestation of this sexually transmitted infection. Despite its significant clinical implications, the lack of distinctive symptoms and the overlap with other ocular conditions often lead to underdiagnosis or misdiagnosis of AIC associated with C. trachomatis infection. Here, we established six distinct C. trachomatis culture cell lines, specifically highlighting the MA104 N*V cell line that exhibited diminished expression of interferon regulatory factor 3 (IRF3) and signal transducer and activator of transcription 1 (STAT1), resulting in reduced interferons. Infected MA104 N*V cells displayed the highest count of intracytoplasmic inclusions detected through immunofluorescence staining, peaking at 48 h postinfection. Subsequently, MA104 N*V cells were employed for clinical screening in adult patients diagnosed with AIC. Among the evaluated cohort of 20 patients, quantitative PCR (qPCR) testing revealed positive results in seven individuals, indicating the presence of C. trachomatis infection. Furthermore, the MA104 N*V cell cultures derived from these infected patients demonstrated successful cultivation and replication of the pathogen, confirming its viability and infectivity. Molecular genotyping identified four distinct urogenital serovars, with serovar D being the most prevalent (4/7), followed by E (1/7), F (1/7), and Ia (1/7). This novel cellular model contributes to studies on C. trachomatis pathogenesis, molecular mechanisms, and host-pathogen interactions both in vitro and in vivo. It also aids in acquiring clinically relevant strains critical for advancing diagnostics, treatments, and vaccines against C. trachomatis., (© The Author(s) 2024. Published by Oxford University Press on behalf of FEMS.)

Aim: To report two atypical inclusion conjunctivitis cases due to Chlamydia trachomatis in young adults., Method: Transcription mediated amplification for C. trachomatis was performed using Aptima Combo 2 Assay (Hologic, Spain)., Results: The first patient was managed as an orbital disorder because he had unilateral location, and ptosis was observed. Orbital nuclear magnetic resonance revealed normal results, and conjunctival biopsy did not indicate significant results. For the second patient, thyroid eye disease was suspected, but the orbital nuclear magnetic resonance revealed normal results. Conjunctival exudate samples were collected and sent to the Microbiology Laboratory where C. trachomatis was confirmed. Both patients demonstrated a great improvement with oral azithromycin 1 g., Conclusion: Inclusion conjunctivitis could present as unspecified unilateral or bilateral chronic conjunctivitis. Thus, suspecting it would be important in order to prevent spread and wasting diagnostic resources., Competing Interests: Declaration of competing interest None declared. Written informed consent was obtained from the patients for publication of this case report and accompanying images. A copy of the written consent is available for review by the Editor-in-Chief of this journal on request., (Copyright © 2022 Institut Pasteur. Published by Elsevier Masson SAS. All rights reserved.)

Trachoma is a blinding disease caused by repeated conjunctival infection with different Chlamydia trachomatis (Ct) genovars. Ct B genovars have been associated with more severe trachoma symptoms. Here, we investigated associations between Ct genovars and bacterial loads in ocular samples from two distinct geographical locations in Africa, which are currently unclear. We tested ocular swabs from 77 Moroccan children (28 with trachomatous inflammation-follicular (TF) and 49 healthy controls), and 96 Sudanese children (54 with TF and 42 healthy controls) with a Ct-specific real-time polymerase chain reaction (PCR) assay. To estimate bacterial loads, Ct-positive samples were further processed by multiplex real-time qPCR to amplify the chromosomal outer membrane complex B and plasmid open reading frame 2 of Ct. Genotyping was performed by PCR-based amplification of the outer membrane protein A gene (~1120 base pairs) of Ct and Sanger sequencing. Ct-positivities among the Moroccan and Sudanese patient groups were 60·7% and 31·5%, respectively. Significantly more Sudanese patients than Moroccan patients were genovar A-positive. In contrast, B genovars were significantly more prevalent in Moroccan patients than in Sudanese patients. Significantly higher Ct loads were found in samples positive for B genovars (598596) than A genovar (51005). Geographical differences contributed to the distributions of different ocular Ct genovars. B genovars may induce a higher bacterial load than A genovars in trachoma patients. Our findings emphasize the importance of conducting broader studies to elucidate if the noted difference in multiplication abilities are genovar and/or endemicity level dependent., Competing Interests: The authors have declared that no competing interests exist.

Trachoma is a devastating neglected tropical disease caused by Chlamydia trachomatis and the leading global cause of infectious blindness. Although antibiotic treatment against trachoma is efficient (SAFE strategy), additional affordable therapeutic strategies are of high interest. Water-filtered infrared A and visible light (wIRA/VIS) irradiation has proven to reduce chlamydial infectivity in vitro and ex vivo. The aim of this study was to evaluate whether wIRA/VIS can reduce chlamydial infection load and/or ocular pathology in vivo, in a guinea pig model of inclusion conjunctivitis. Guinea pigs were infected with 1 × 10 6 inclusion-forming units/eye of Chlamydia caviae via the ocular conjunctiva on day 0. In infected animals, wIRA/VIS irradiation (2100 W/m 2 ) was applied on day 2 (single treatment) and on days 2 and 4 (double treatment) post-infection (pi). wIRA/VIS reduced the clinical pathology score on days 7 and 14 pi and the conjunctival chlamydial load on days 2, 4, 7, and 14 pi in comparison with C. caviae-infected, not irradiated, controls. Furthermore, numbers of chlamydial inclusions were decreased in wIRA/VIS treated C. caviae-infected guinea pigs on day 21 pi compared to C. caviae-infected, non-irradiated, controls. Double treatment with wIRA/VIS (days 2 and 4 pi) was more efficient than a single treatment on day 2 pi. wIRA/VIS treatment did neither induce macroscopic nor histologic changes in ocular tissues. Our results indicate that wIRA/VIS shows promising efficacy to reduce chlamydial infectivity in vivo without causing irradiation related pathologies in the follow-up period. wIRA/VIS irradiation is a promising approach to reduce trachoma transmission and pathology of ocular chlamydial infection., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2020 Elsevier B.V. All rights reserved.)

Nucleic acid amplification tests are increasingly used to detect ocular chlamydia infection in trachoma research and programs. To evaluate the reliability of Chlamydia trachomatis detection by the Abbott RealTime CT/NG assay (Abbott Molecular, Inc., Des Plaines, IL) on the m2000 platform, three conjunctival samples were collected from each of 200 children aged 0-9 years in Ethiopia: two from the right eye and one from the left eye. Four aliquots were processed for each child: two from the first right eye sample, one from the second right eye sample, and one from the left eye sample. Sixty-nine swabs were processed in a U.S. laboratory and 131 in an Ethiopian laboratory. Intra-class correlation coefficients (ICCs) were high when comparing two aliquots from the same swab (ICC ranged from 0.96 to 0.99), two separate swabs from the right eye (0.89-0.91), and one right and one left eye swab (0.87-0.89), indicating reliable chlamydial load assessment across different samples and laboratory settings.

We report an unusual case of an 11-day-old neonate presenting with haemolacria on a background of sticky conjunctival discharge. This was secondary to Chlamydia pseudomembranous conjunctivitis which responded well to systemic erythromycin. Early appropriate treatment is important to prevent progression of the ophthalmic infection, which could lead to blindness, and to prevent other manifestations of neonatal chlamydial infection, particularly pneumonia, which could be fatal. Management also includes treating the mother and educating about sexually transmitted infections., Competing Interests: Competing interests: None declared., (© BMJ Publishing Group Limited 2020. No commercial re-use. See rights and permissions. Published by BMJ.)

Background: Although Chlamydia trachomatis infection is common in pregnant women, such infections are rarely encountered in infants. To clarify the recognition of C. trachomatis infections in infants, we analyzed symptoms and signs of perinatally acquired chlamydial infection, together with its long-term prognosis in a large population-based patient series., Methods: A search through 2 national health registries covering 1996-2011, a cohort of 933,823 births, yielded 124 children with a microbiologically confirmed C. trachomatis infection. We then reviewed copies of the children's medical records up to 16 years of age., Results: One-third of the infants with chlamydial conjunctivitis (33/110) had spontaneous blood-stained discharge from the infected eye. The infants with C. trachomatis lower respiratory tract infection were mostly afebrile (30/32), and 15/32 of them had wheezing. Staccato cough was not recorded in any of the infants. The median diagnostic delay from the onset of the symptoms was 13 (range 4-374) days for conjunctivitis and 25 (range 10-149) days for lower respiratory tract infection. One neglected child developed bilateral corneal scars because of an untreated C. trachomatis infection., Conclusions: Blood-stained discharge was a typical finding in C. trachomatis conjunctivitis. The C. trachomatis-infected infants had severe and prolonged symptoms, but long-term consequences were rare. The diagnostic delay was long, especially among the infants with a C. trachomatis lower respiratory tract infection.

The objective of the present study was to determine the frequency and age distribution of different Chlamydia trachomatis (CT) genotypes causing ophthalmia neonatorum (ON) in Hungary. Using CT specific PCR, we tested 76 conjunctival samples from symptomatic infants up to 3 months old in the National Centre for Epidemiology, Budapest between 2008 and 2016. CT tested positive in 30 of 76 conjunctival samples (39.5 %). The sequencing of the positive samples was successful in every case but one, and resulted in 48 % dominance for genotype E (14/29), followed by 24 % for genotype G (7/29), 10 % for J (3/29), 6.9 % for K and F (2/29), and 3.4 % for H (1/29). CT must still be regarded as a common pathogen causing ON in Hungary. Routine screening and treatment of pregnant women can be recommended to prevent these conditions. Chronic ON cases can be reduced by early diagnosis. Further research is needed to explain the dominance of genotypes E and G.

Perinatal transmission of Chlamydia trachomatis (CT) and Neisseria gonorrhoeae (GC) can result in conjunctivitis in infants. We examined national rates of reported CT/GC conjunctivitis among infants. Surveillance of these infections is heavily affected by the completeness of reported data on specimen source and age. Alternative data sources should be evaluated.

Conjunctivitis is a frequently diagnosed disease, usually caused by a virus. A less well-known cause is a chlamydia infection. This may result in missed diagnoses, delay of treatment and several complications. We present two cases of a persistent, therapy-resistant conjunctivitis in patients who were over 70 years of age. One patient had conjunctival follicles, characteristic of chlamydia conjunctivitis. The polymerase chain reaction tests of conjunctival samples from both patients were positive for chlamydia. Both patients and their sexual partners were treated with oral azithromycin. There was a treatment delay in both cases due to late recognition which was partially due to the older age of the patients. These cases illustrate that when a patient presents with persistent, therapy-resistant conjunctivitis, particularly if conjunctival follicles are present, chlamydial conjunctivitis should be considered and conjunctival swabs should be taken, no matter what the age of the patient.

Background: Recent findings indicated that a correlation between genomic % AT and genome size within strains of microbial species was predominantly associated with the uptake of foreign DNA. One species however, Chlamydia trachomatis, defied any explanation. In the present study 79 fully sequenced C. trachomatis genomes, representing ocular- (nine strains), urogenital- (36 strains) and lymphogranuloma venereum strains (LGV, 22 strains), in three pathogroups, in addition to 12 laboratory isolates, were scrutinized with the intent of elucidating the positive correlation between genomic AT content and genome size., Results: The average size difference between the strains of each pathogroup was largely explained by the incorporation of genetic fragments. These fragments were slightly more AT rich than their corresponding host genomes, but not enough to justify the difference in AT content between the strains of the smaller genomes lacking the fragments. In addition, a genetic region predominantly found in the ocular strains, which had the largest genomes, was on average more GC rich than the host genomes of the urogenital strains (58.64% AT vs. 58.69% AT), which had the second largest genomes, implying that the foreign genetic regions cannot alone explain the association between genome size and AT content in C. trachomatis. 23,492 SNPs were identified for all 79 genomes, and although the SNPs were on average slightly GC rich (~47% AT), a significant association was found between genome-wide SNP AT content, for each pathogroup, and genome size (p < 0.001, R (2) = 0.86) in the C. trachomatis strains., Conclusions: The correlation between genome size and AT content, with respect to the C. trachomatis pathogroups, was explained by the incorporation of genetic fragments unique to the ocular and/or urogenital strains into the LGV- and urogential strains in addition to the genome-wide SNP AT content differences between the three pathogroups.

Objective: Chlamydia trachomatis conjunctivitis may present with extended symptoms, and it can have social ramifications as a sexually transmitted disease. For appropriate therapy, C. trachomatis conjunctivitis should be diagnosed definitively. This study presents the verification of nucleic acid amplification testing (NAAT; Gen-Probe Aptima Combo 2 assay) for detection of C. trachomatis ribosomal RNA (rRNA) from direct ocular samples., Design: Retrospective laboratory verification study., Subjects: Patients with infectious conjunctivitis., Methods: A battery of 25 true-positive specimens (direct ocular specimens from patients with symptoms consistent with C. trachomatis conjunctivitis and with previously demonstrated positive polymerase chain reaction [PCR] results for C. trachomatis DNA by Roche Amplicor) and 25 true-negative specimens (direct ocular specimens with culture-positive results for herpes simplex virus [n = 5], adenovirus [n = 5], Haemophilus influenzae [n = 5], and Streptococcus pneumoniae [n = 5]), and transport medium (n = 5) were tested for C. trachomatis rRNA by NAAT. These true-negative specimens have differential etiologic agents of infectious conjunctivitis. The 25 C. trachomatis specimens with PCR-positive results (obtained May 1994-May 2012) and 20 true-negative infectious ocular specimens (obtained December 2008-August 2013) were collected with soft-tipped applicators and placed in transport medium. All excess specimens were stored at -80°C. All samples were centrifuged at 13,000 rpm for 1 hour at 6°C. For each sample, using the Aptima Unisex collection blue swab, a specimen was collected from the conical apex of the storage tube where a pellet was formed. The Aptima Unisex collection swab was placed in a tube of Aptima swab transport medium for testing. All samples were tested in duplicate., Main Outcome Measures: Detection of C. trachomatis rRNA., Results: Of 25 true-positive samples, 24 (96%) were positive by NAAT, whereas 25 of 25 true-negative samples (100%) showed negative results. The sensitivity, specificity, positive predictive value, negative predictive value, and efficiency were determined to be 96%, 100%, 100%, 96%, and 98%, respectively., Conclusions: The detection of C. trachomatis in ocular specimens by NAAT was verified for laboratory diagnosis. The test will be evaluated prospectively to determine future test performance precisely., (Copyright © 2015 American Academy of Ophthalmology. Published by Elsevier Inc. All rights reserved.)

Gonococcal conjunctivitis is a rare infection induced by Neisseria gonorrhoeae and it usually manifests as a hyperacute purulent conjunctivitis. Ocular access of the infectious secretion during sexual intercourse is the way of transmission among adults. Inclusion conjunctivitis caused by the serovars D-K of Chlamydia trachomatis also affects the sexually active population. Authors present a case of a 33-year-old homosexual man who was treated for late latent syphilis formerly. Clinical symptoms were yellow purulent discharge for 3 weeks without any urological or upper respiratory tract symptoms. Conjunctival Neisseria gonorrhoeae and Chlamydia trachomatis infection was identified using cultures and polymerase chain reaction; pharyngeal swab culture and polymerase chain reaction showed positive results for both pathogens. The patient was probably under influence of party drugs at the time of sexual abuse when he became infected. After parenteral and oral cephalosporin and azithromycin therapy the patient had complete recovery within three weeks.

Introduction: Implementation of an in-house polymerase chain reaction (PCR) multiplex assay by West of Scotland Specialist Virology Centre to improve sample processing means all viral eye swabs are now routinely tested for Adenovirus, Herpes simplex, Varicella and Chlamydia. Concern was raised regarding subsequent management and sexual health attendance for Chlamydia-positive patients identified in eye casualty., Methods: A retrospective review of virology results identified 76 Chlamydia-positive patients from 1914 eye swabs (4%) from May 2007 to April 2008. Of these results, 12 originated from Glasgow eye casualty and available clinical notes were cross-referenced with the sexual health network (Sandyford)., Results: Identified issues included no documentation of implications of testing, poor communication of positive results and poor referral pathways to sexual health for assessment; all leading to inadequate management. A shared care network was created to address these issues. A designated sexual health advisor was identified to improve sexual health referral, specialist assessment, standardised management and contact tracing. Re-audit showed more consistent follow-up., Conclusion: New PCR technology has resulted in a shared care approach to address corresponding implications of testing. Effective communication with a structured protocol and a central point of contact has improved follow-up and ensures appropriate best practice management of chlamydial conjunctivitis.

Introduction: Chlamydia trachomatis causes many infections, including eye infections. They manifest as inclusion conjunctivitis and trachoma. The agent is transmitted by dirty hands, eyeliners, medical instruments and via swimming-pool water or, in neonates, by passage through an infected birth canal. Due to the nonspecific clinical features at the beginning of the infection and delayed application of symptomatic, anti-allergic and non-specific antibiotic therapy, Chlamydia aetiology is usually established only after laboratory diagnosis in the chronic stage of infection., Objective: Determining the frequency of Chlamydia trachomatis antigen in conjunctival and genital samples of adult patients with chronic conjunctivitis in Nisava district., Methods: Our retrospective study was carried out on 116 patients (63 female and 53 male) with clinical signs and symptoms of chronic conjunctivitis. Chlamydia trachomatis antigen was detected by a direct immunofluorescence test with labelled monoclonal antibodies., Results: From a total of 116 examined patients in 37 patients Chlamydia trachomatis antigen was detected; 17 female and 20 male. Thirty-three of the patients had a bilateral infection and four unilateral. Among 24 patients who were also tested for Chlamydia trachomatis antigen collected by ocular and genital swabs, 19 had conjunctivitis associated with urethritis/vaginitis., Conclusion: The studied group of patients showed that the common cause of the chronic conjunctivitis were bacteria, but predominantly Chlamydia trachomatis. In most cases Chlamydia infection occurred bilaterally. The majority of patients had eye Chlamydia infection associated with genital Chlamydia infection. There was no statistically significant difference in the presence of the disease regarding gender.

In order to study the interaction of variants in in vivo infection, we employed an azithromycin-resistant mutant (AZ(2)) and its wild-type parent (SP(6)) in the guinea pig model of Chlamydia caviae conjunctival infection. When each strain was inoculated individually into conjunctiva, both attained the same level of growth, but AZ(2) elicited less pathology. However, when equal numbers of the two strains were inoculated together into the guinea pig conjunctiva, SP(6) produced a significantly greater number of inclusion-forming units than AZ(2), and the pathology reflected that of a SP(6) monoinfection. The goal of this study was to further characterize the dynamics of concomitant infection of these two distinct variants, with particular emphasis on the impact of the host response on the in vivo growth of each organism and the development of pathology. Animals infected with AZ(2) had reduced conjunctival infiltration with CD45(+) cells and neutrophils as well as a reduced interleukin-8 (IL-8) response. Gene expression of gamma interferon (IFN-γ), tumor necrosis factor alpha (TNF-α), CCL2, and CCL5 was also significantly lower in AZ(2)-infected animals. The lower inflammatory response induced by AZ(2) was associated with its decreased ability to activate NF-κB via Toll-like receptor 2 (TLR2). In general, the inflammatory response in animals infected with both variants was greater than in infection with AZ(2) alone, resulting in lower numbers of AZ(2) than those of SP(6) in the mixed infection. Our results suggest that the ability to elicit an inflammatory response is an important factor in the dynamics of mixed infection with strains that display different pathological phenotypes.

Until the present time, ophthalmic chlamydiasis has been generally associated with diseases of auxiliary organs of the eye and its anterior segment: conjunctivitis and iridocyclitis. The morphological substrate of eye posterior segment lesion caused by C. trachomatis and C. pneumonia was studied in this investigation. The pathomorphological pattern characteristic of chlamydia-induced rabbit retinal and vitreous body lesions is composed of vitreoretinal lymphocyte-macrophageal infiltrations of varying intensity, posterior hyaloid membrane detachment, peripheral foci or folding of the retina, impaired nuclei of photoreceptors and bipolar neurons, pigment epithelial damage occurring in different concurrences in relation to the species of a causative agent and the clinical picture.

Azithromycin (AZM) is a major drug used in the treatment and prophylaxis of infections caused by Chlamydia, yet no significant clinical resistance has been reported for these obligate intracellular bacteria. Nevertheless, spontaneous AZM resistance (Azm(r)) arose in vitro at frequencies ranging from 3 x 10(-8) to 8 x 10(-10) for clonal isolates of Chlamydia caviae, which is a natural pathogen of guinea pigs. Sequencing of the unique 23S rRNA gene copy in 44 independent Azm(r) isolates identified single mutations at position A(2058) or A(2059) (Escherichia coli numbering system). While SP(6)AZ(1) (A(2058)C) and SP(6)AZ(2) (A(2059)C) Azm(r) mutants showed growth defects in cell culture and were less pathogenic in the guinea pig ocular infection model than in the parent SP(6), the three isogenic C. caviae isolates grew equally well in the animal. On the other hand, coinoculation of the C. caviae parent strain with one of the Azm(r) strains was detrimental for the mutant strain. This apparent lack of association between pathology and bacterial load in vivo showed that virulence of the two Azm(r) mutants of C. caviae was attenuated. While chlamydial growth in vitro reflects the ability of the bacteria to multiply in permissive cells, survival in the host is a balance between cellular multiplication and clearance by the host immune system. The obligate intracellular nature of Chlamydia may therefore limit emergence of resistance in vivo due to the strength of the immune response induced by the wild-type antibiotic-sensitive bacteria at the time of antibiotic treatment.

While much is known about the attachment of the chlamydiae to the host cell and intracellular events during the developmental cycle, little is known about the mechanism(s) by which elementary bodies exit the cell. In this report, we use the guinea-pig conjunctival model of Chlamydia caviae infection to present in vivo ultrastructural evidence supporting two mechanisms for release of chlamydiae from the mucosal epithelia. Four days after infection, histopathologic observation shows an intense infiltration of polymorphonuclear leukocytes (PMN) in the conjunctival epithelium. Using transmission electron microscopy, a gradient-directed PMN response to chlamydiae-infected epithelial cells was observed. As PMN infiltration intensifies, epithelial hemidesmosome/integrin/focal adhesion adherence with the basal lamina is disconnected and PMNs literally lift off and release infected superficial epithelia from the mucosa. Many of these infected cells appear to be healthy with intact microvilli, nuclei, and mitochondria. While lysis of some infected cells occurs with release of chlamydiae into the extracellular surface milieu, the majority of infected cells are pushed off the epithelium. We propose that PMNs play an active role in detaching infected cells from the epithelium and that these infected cells eventually die releasing organisms but, in the process, move to new tissue sites via fluid dynamics.

Background: Trachoma, caused by Chlamydia trachomatis (Ct), is the leading infectious cause of blindness. Sequence-based analysis of the multiple strains typically present in endemic communities may be informative for epidemiology, transmission, response to treatment, and understanding the host response., Methods: Conjunctival and nasal samples from a Gambian community were evaluated before and 2 months after mass azithromycin treatment. Samples were tested for Ct by Amplicor, with infection load determined by quantitative PCR (qPCR). ompA sequences were determined and their diversity analysed using frequency-based tests of neutrality., Results: Ninety-five of 1,319 (7.2%) individuals from 14 villages were infected with Ct at baseline. Two genovars (A and B) and 10 distinct ompA genotypes were detected. Two genovar A variants (A1 and A2) accounted for most infections. There was an excess of rare ompA mutations, not sustained in the population. Post-treatment, 76 (5.7%) individuals had Ct infection with only three ompA genotypes present. In 12 of 14 villages, infection had cleared, while in two it increased, probably due to mass migration. Infection qPCR loads associated with infection were significantly greater for A1 than for A2. Seven individuals had concurrent ocular and nasal infection, with divergent genotypes in five., Conclusions: The number of strains was substantially reduced after mass treatment. One common strain was associated with higher infection loads. Discordant genotypes in concurrent infection may indicate distinct infections at ocular and nasal sites. Population genetic analysis suggests the fleeting appearance of rare multiple ompA variants represents purifying selection rather than escape variants from immune pressure. Genotyping systems accessing extra-ompA variation may be more informative.

Purpose: To compare the performance of polymerase chain reaction versus conventional methods (cell culture and direct immunofluorescent assay) in diagnosing neonatal chlamydial conjunctivitis and their correlations to the severity of conjunctivitis., Methods: Consecutive cases of neonatal conjunctivitis were recruited over a year. Both eyes were clinically graded according to the severity of conjunctivitis and investigated using the three aforementioned chlamydial tests. Neonatal chlamydial conjunctivitis was assumed if one of these three tests was positive and there was clinical improvement after treatment. Sensitivity and specificity of each of the tests were analyzed., Results: Three hundred sixty-eight sets of chlamydial tests were done for 184 neonates. The percentage of positive results was 93.8% and 71.9% for polymerase chain reaction and conventional methods, respectively. Using positive results in either cell culture or direct immunofluorescent assay as a standard to diagnose neonatal chlamydial conjunctivitis, the sensitivity and specificity of polymerase chain reaction were 92.0% and 97.7%, respectively. If we used polymerase chain reaction as a standard, the sensitivity and specificity of cell culture were 73.3% and 99.7%, respectively. A discrepancy was noted in the number of positive results between polymerase chain reaction and conventional methods in milder disease., Conclusions: Polymerase chain reaction might have a higher sensitivity and similar specificity in diagnosing neonatal chlamydial conjunctivitis compared to conventional methods, and it has an additional advantage as a diagnostic tool in mild disease.

Objectives: Chlamydia trachomatis infections, in the context of extreme poverty, may trigger trachoma. Because the levels of C. trachomatis eye infections in Mexico are unknown, this study sought to determine if C. trachomatis was present in the conjunctiva of children living in three poor, rural areas of the country., Methods: Clinical diagnosis of conjunctival follicles in children was conducted during the 2004 visual acuity assessment campaigns in rural areas of the states of Chiapas, Oaxaca, and Zacatecas. C. trachomatis detection was carried out by sampling the children with follicles and examining the specimens after Giemsa or microimmunofluorescence (MIF) staining., Results: A total of 941 children from 6 to 12 years of age were examined in 2004. Of the 484 in Chiapas, 30% were found to have follicles; of the 181 in Zacatecas, 22%; and of the 276 in Oaxaca, 42%. C. trachomatis was detected at levels ranging between 2% and 5%; positive by Giemsa in 4.5% of the children with follicles, and by MIF in 15.5%., Conclusions: Considering that the chlamydiae sampling procedures and detection methods used in this study were not the most sensitive, the results underestimate the chlamydial eye infections and represent a conservative assessment of a potential risk for preventable visual impairment. Because C. trachomatis was detected here at levels similar to those reported for low-endemic trachoma areas, health authorities should be prepared to implement appropriate measures should it be confirmed that the visual health of Mexico's children is at risk.

Purpose: The World Health Organization recommends mass treatment of trachoma-hyperendemic communities, but there are scant empiric data on the number of rounds of treatment that are necessary for sustainable reductions. The rates of active trachoma and infection with C. trachomatis were determined in a community 3.5 years after two rounds of mass treatment with azithromycin., Methods: Maindi village in Tanzania received a first round of mass treatment with azithromycin after a baseline survey for trachoma and infection. All residents aged 6 months and older were offered single-dose treatment with azithromycin (excluding pregnant women with no clinical trachoma, who were offered topical tetracycline). The residents were followed over an 18-month period, and, according to similar treatment criteria, were offered retreatment at 18 months. Five years after baseline (3.5 years after the second round of mass treatment), a new census and survey of current residents for trachoma and infection was conducted. Children are the sentinel markers of infection and trachoma in communities, so data are presented specifically for ages 0 to 7 years (preschool age) and 8 to 16 years., Results: Treatment coverage was above 80% for all ages in the first round, and highest (90%) in preschool-aged children. Second-round coverage was lower, <70%, and 70% in preschool-aged children. At 5 years, trachoma rates were still lower than baseline, ranging from 45% in those aged 0 to 3 years to 8% in those aged 11 to 15 years (compared with 81% and 39% at baseline, respectively). Infection rates at baseline ranged from 71% to 57%, but were 27% to 17% at 5 years after two rounds of mass treatment. At 5 years, there were no differences in trachoma or infection rates, when comparing new residents who came after the second mass treatment with those who had been resident in the village during both rounds (P > 0.05). Infection rates were lower in those who had been treated twice or at 18 months than in those treated only at baseline or never treated., Conclusions: Although mass treatment appears to be associated with lower disease and infection rates in the long term, trachoma and C. trachomatis infection were not eliminated in this trachoma hyperendemic village 3.5 years after two rounds of mass treatment. Continued implementation of the SAFE strategy in this environment is needed.

The majority of cases of chlamydial conjunctivitis are thought to result from autoinoculation by the patient of infected genital secretions from themselves or their sexual partners. We noted that some patients had developed symptoms following direct ejaculation into the affected eye. We describe four cases of chlamydial conjunctivitis following ejaculation of semen directly into the eye, which have not been previously described. In only one case was chlamydia detected in the genital tract. In three cases, there was no evidence of genital chlamydial infection; the sources of the eye infection being either from infected genital material of their sexual partners transferred by hands to the eyes, or more likely from direct ejaculate inoculation. It is likely that this mode of transmission is underestimated as a history of ejaculation into the conjunctiva is not normally asked for.

Chlamydia trachomatis ocular and urogenital infections represent major public health problems, whereas Chlamydophila pneumoniae is a common aetiological agent of community-acquired pneumonia. The obligate intracellular lifestyle of these established pathogens poses challenges to both their diagnosis and treatment. Tetracyclines, macrolides and quinolones remain the antimicrobials of choice for the treatment of infections due to Chlamydiaceae.

In 1911, Lindner and colleagues identified intracytoplasmic inclusions in infants with a nongonococcal form of ophthalmia neonatorum called inclusion conjunctivitis of the newborn (ICN). Mothers of affected infants were found to have inclusions in their cervical epithelial cells, fathers of such infants had inclusions in their urethral cells, and the epidemiology of sexually transmitted chlamydial infections was revealed. Fifty years later, chlamydial isolation procedures were developed, and studies again demonstrated Chlamydia trachomatis as an etiology of ICN and the female birth canal as the reservoir. In the late 1970s, a report by Beem and Saxon described respiratory tract colonization and a distinct pneumonia syndrome in infected infants. Genital chlamydial infection is recognized as the world's most common sexually transmitted disease, with estimates of greater than 4 million new infections occurring annually in the United States. Although most C. trachomatis infections in men and women are asymptomatic, infection can lead to severe reproductive complications in women. The high prevalence in women of child-bearing age results in exposure of an estimated 100,000 neonates in the United States annually. Many of these infants develop conjunctivitis, pneumonia, or both in the first few months of life. Clinical features, diagnosis, treatment, and approaches to prevention of conjunctivitis and pneumonia in the newborn and young infant are reviewed here. Appropriate testing for chlamydial infection in a pediatric victim of sexual assault and the implications of identifying C. trachomatis in suspected cases of childhood sexual abuse also are reviewed.

Antibiotics are one of four arms of the SAFE strategy for the control of trachoma, an eye infection that is responsible for more cases of blindness than any condition other than cataract. The evidence for the use of topical tetracycline and oral tetracycline, doxycycline, erythromycin, cotrimoxazole and azithromycin in trachoma are reviewed here and a number of issues are nominated as research and policy priorities.

Purpose: The aim of the study was to compare the efficacy and safety of azithromycin and doxycycline in the treatment of chlamydial conjunctivitis in adults., Design: An open, randomized clinical trial., Methods: Seventy-eight adult patients with incluson conjunctivitis were enrolled in this multicenter clinical study. Patients with chlamydial conjunctivitis as indicated by a positive direct fluorescent antibody (DFA) test or cell culture were randomized to receive a single 1-g dose of azithromycin or doxycycline, 100 mg twice daily for 10 days. A conjuctival swab for cell culture was obtained from all patients immediately before the treatment for subsequent confirmation of the presence of chlamydial infection in the central laboratory. Control examinations were performed 10 to 12 days and 4 to 6 weeks after the treatment initiation. Clinical and bacteriological responses to the treatment were evaluated at the last visit. The occurrence and frequency of adverse events were analyzed as well., Results: Of 78 patients enrolled, 51 completed the study and were evaluated for efficacy. The main reasons for withdrawal were lack of confirmation of the presence of chlamydial infection by the central laboratory and failure to attend the follow-up visit. Eradication of C. trachomatis was achieved in 23 of 25 (92%) patients treated with azithromycin and in 25 of 26 (96%) patients treated with doxycycline. Clinical cure was observed in 15 (60%) and 18 (69%) patients treated with azithromycin and doxycycline, respectively. Both drugs were equally well tolerated., Conclusions: A single 1-g azithromycin therapy was as effective as standard 10-day treatment with doxycycline (100 mg twice daily) in the treatment of adult inclusion conjunctivitis.

Chlamydiae cause a spectrum of diseases in multiple organ systems, and chlamydial infections of the eye lead to sequelae varying from mild conjunctivitis to blindness. This paper reviews current concepts in the pathogenesis and management of ocular chlamydial infections. Trachoma, the leading cause of preventable blindness in the world, is compared with other ocular chlamydial diseases to underscore key concepts in chlamydial pathogenesis. Emerging treatment strategies are discussed in the context of chlamydial pathogenesis and the World Health Organization initiative to eradicate trachoma by 2020.

Infection with an obligate intracellular bacterium, the Chlamydia trachomatis lymphogranuloma venereum (LGV/L2) strain or the guinea pig inclusion conjunctivitis serovar of Chlamydia psittaci, leads to apoptosis of host cells. The apoptosis is not affected by a broad-spectrum caspase inhibitor, and caspase-3 is not activated in infected cells, suggesting that apoptosis mediated by these two strains of Chlamydia is independent of known caspases. Overexpression of the proapoptotic Bcl-2 family member, Bax, was previously shown to induce caspase-independent apoptosis, and we find that Bax is activated and translocates from the cytosol to the mitochondria in C. psittaci-infected cells. C. psittaci-induced apoptosis is inhibited in host cells overexpressing Bax inhibitor-1 and is inhibited through overexpression of Bcl-2, which blocks both caspase-dependent and -independent apoptosis. As Bax and mitochondria are ideally located to sense stress-related metabolic changes emanating from the interior of an infected cell, it is likely that Bax-dependent apoptosis may also be observed in cells infected with other intracellular pathogens.

During two years (1997-1999) an investigation of possible infections of chlamydial etiology in outpatients with follicular conjunctivitis was carried out, through the use of specific assays. Fifty-seven selected patients with presumptive inclusion conjunctivitis were diagnosed by means of ophthalmoscopic examination and bilateral tarsal-conjunctiva swabbing for microorganisms. The possible presence of Chlamydia trachomatis was tested by immunofluorescence microscopy and isolation in cell culture of McCoy line. Of the 57 conjunctivitis patients screened, 37 (65%) proved to be positive by cell culture (CC) and 27 (47%) by direct immunofluorescence (IFD). A good agreement between the two assays was observed, where the CC was more sensitive than IFD. Of these 37 patients with chlamydial conjunctivitis, 23 (62%) were women, with over one-third of them ranging in age from 45 to 65 years. Their clinical records revealed an evolution period of 1 to 12 months. Eighteen (78%) of these women reported previous genital pathology, while 4 (29%) of the 14 men had a history of urethritis by Chlamydia trachomatis. A high frequency of follicular conjunctivitis by Chlamydia (65%) in the screened patients was observed, without any evidence of urogenital signs and symptoms at the moment of the study.

Purpose: To evaluate type-specific antichlamydial antibody IgG in children with chronic follicular conjunctivitis., Methods: A total of 90 serum samples from juvenile patients with chronic follicular conjunctivitis were collected in the Southeastern Anatolian region of Turkey where trachoma is still endemic. These samples were investigated regarding Chlamydia trachomatis pooled serotype-specific IgG by using the micro-immunofluorescence test. A titer of 1/32 or higher was considered positive., Results: Specific IgG seropositivity to Chlamydia trachomatis titer was found in 33 (36.1%) of the 90 subjects. A higher titer was observed frequently in the serotypes pooled in BDE (21 subjects), CHIJ (10 subjects), and FGK (2 subjects), respectively., Conclusions: The presence of antichlamydial antibodies in blood should always be interpreted in accordance with the history of the patient, the clinical picture, and the course of the disease. In the diagnosis of Chlamydia trachomatis infections in a patient with chronic follicular conjunctivitis, not only genus-specific antibodies but also the presence of the subspeciespecific antibodies should be investigated.

Background: Chlamydia trachomatis is one of the most common identifiable infectious agents in neonatal conjunctivitis. It also causes pneumonitis, that is preceded by conjunctivitis in one third of cases., Aim: To asses the prevalence of Chlamydia trachomatis in newborns with conjunctivitis., Patients and Methods: In 162 newborns, coming from 14 Primary Health Centers from Santiago de Chile, C. trachomatis was detected by indirect fluorescence and two polymerase chain reaction (PCR 1 and 2), which amplified different sequences from the common endogenous plasmid. Those patients with positive indirect fluorescence and PCR 2 were defined as infected:, Results: The prevalence of C. trachomatis was 8%, and the distribution of the positive cases was similar in the different Health Centers. Other isolates were: S. aureus (9.8%), S. pneumoniae (8%), S. viridans (6.2%) y H. influenzae (5.5%)., Conclusions: The prevalence of C. trachomatis in neonatal conjunctivitis in Chile is similar to that of developed countries. Therefore, C. trachomatis should be considered in the election of antimicrobials for the treatment of neonatal conjunctivitis, to avoid ocular and respiratory complications.

The authors report the isolation and molecular characterization of a bacteriophage, &phi;CPG1, which infects CHLAMYDIA: psittaci strain Guinea pig Inclusion Conjunctivitis. Purified virion preparations contained isometric particles of 25 nm diameter, superficially similar to spike-less members of the &phi;X174 family of bacteriophages. The single-stranded circular DNA genome of &phi;CPG1 included five large ORFs, which were similar to ORFs in the genome of a previously described CHLAMYDIA: bacteriophage (Chp1) that infects avian C. psittaci. Three of the ORFs encoded polypeptides that were similar to those in a phage infecting the mollicute Spiroplasma melliferum, a pathogen of honeybees. Lesser sequence similarities were seen between two ORF products and the major capsid protein of the &phi;X174 coliphage family and proteins mediating rolling circle replication initiation in phages, phagemids and plasmids. Phage &phi;CPG1 is the second member of the genus CHLAMYDIAMICROVIRUS:, the first to infect a member of a CHLAMYDIA: species infecting mammals. Similarity searches of the nucleotide sequence further revealed a highly conserved (75% identity) 375 base sequence integrated into the genome of the human pathogen Chlamydia pneumoniae. This genomic segment encodes a truncated 113 residue polypeptide, the sequence of which is 72% identical to the amino-terminal end of the putative replication initiation protein of &phi;CPG1. This finding suggests that C. pneumoniae has been infected by a phage related to &phi;CPG1 and that infection resulted in integration of some of the phage genome into the C. pneumoniae genome.

The infectious cycle of phiCPG1, a bacteriophage that infects the obligate intracellular pathogen, Chlamydia psittaci strain Guinea Pig Inclusion Conjunctivitis, was observed using transmission electron microscopy of phage-hyperinfected, Chlamydia-infected HeLa cells. Phage attachment to extracellular, metabolically dormant, infectious elementary bodies and cointernalisation are demonstrated. Following entry, phage infection takes place as soon as elementary bodies differentiate into metabolically active reticulate bodies. Phage-infected bacteria follow an altered developmental path whereby cell division is inhibited, producing abnormally large reticulate bodies, termed maxi-reticulate bodies, which do not mature to elementary bodies. These forms eventually lyse late in the chlamydial developmental cycle, releasing abundant phage progeny in the inclusion and, upon lysis of the inclusion membrane, into the cytosol of the host cell. Structural integrity of the hyperinfected HeLa cell is markedly compromised at late stages. Released phage particles attach avidly to the outer leaflet of the outer membranes of lysed and unlysed Chlamydiae at different stages of development, suggesting the presence of specific phage receptors in the outer membrane uniformly during the chlamydial developmental cycle. A mechanism for phage infection is proposed, whereby phage gains access to replicating chlamydiae by attaching to the infectious elementary body, subsequently subverting the chlamydial developmental cycle to its own replicative needs. The implications of phage infection in the context of chlamydial infection and disease are discussed.

Background: Acute follicular conjunctivitis is a clinical diagnosis common to multiple etiologies, of which chlamydial infection requires specific antibiotic treatment., Purpose: This prospective study was designed to evaluate Chlamydia trachomatis as the cause of acute follicular conjunctivitis by two sensitive tests: direct enzyme-linked immunosorbent assay (ELISA) and polymerase chain reaction (PCR)., Methods: Conjunctival scrapings from patients presented with untreated acute follicular conjunctivitis were examined by ELISA and PCR, and patients were followed up for prolongation of the disease course., Results: All 36 consecutive patients presented with acute follicular conjunctivitis were negative for Chlamydia trachomatis by ELISA and PCR. None of the patients had a prolonged course of more than 4 weeks or required treatment with systemic antibiotics as would be expected from chlamydial infection., Conclusions: Chlamydia trachomatis was probably not responsible for the acute follicular conjunctivitis in this series, and ELISA and PCR may not be cost effective for evaluation of acute follicular conjunctivitis due to chlamydial infection. Further evaluation of the cost effectiveness of these tests is required in chronic follicular conjunctivitis.

Analysis of scrapings off the conjunctiva of patients with suspected adenoviral infection (n = 60) and ophthalmochlamydiasis (n = 132) by different diagnostic tests showed good prospects of using DNA hybridization: the efficacy of detecting adenoviral diseases of the eyes was 35% improved in comparison with the fluorescent antibody method with polyclonal serum and the diagnosis of ophthalmochlamydiasis was 22.8% improved in comparison with enzyme immunoassay.

The BioStar OIA Chlamydia test (BioStar, Inc., Boulder, Colo.) is a novel immunoassay system that uses changes in reflection of light to directly detect chlamydial antigen in clinical specimens. We compared the optical immunoassay (OIA) with culture for detecting Chlamydia trachomatis in ocular specimens from infants with suspected chlamydial conjunctivitis. We initially performed a retrospective evaluation, testing 152 ocular specimens previously collected for culture with the OIA. The sensitivity and specificity were 94.2 and 97%, respectively. A subsequent prospective study evaluating 37 ocular specimens from infants with suspected C. trachomatis conjunctivitis revealed a sensitivity and specificity of 100 and 92.6%, respectively.

A molecular biological method of detecting and serotyping Chlamydia trachomatis (C. trachomatis) directly from conjunctival specimens by polymerase chain reaction (PCR) was developed and applied to the diagnosis of inclusion conjunctivitis. We amplified 1.2 kbp DNA fragments of ompA gene from 15 reference strains of C. trachomatis by two-step PCR using two pairs of primers. Restriction fragment length polymorphism analysis using a combination of three endonucleases (HinfI, HindIII and HhaI) completely differentiated 13 of the 15 serovars; the exceptions were B and Ba. We then used this method for 18 strains of C. trachomatis isolated from Japanese patients with inclusion conjunctivitis, serotyping them into six groups: D (5/18), G (5/18), E (3/18), H (2/18), F (1/18), and K (1/18). In our comparison of cell culture isolation with PCR analysis of 38 conjunctival swabs from 35 patients in Sapporo with follicular conjunctivitis, 8 were positive in culture isolation and were also positive in two-step PCR. Twenty-five of 26 strains (the 18 isolated strains and 8 strains amplified by two-step PCR) were genotyped to D, G, H, E, F, and K. One isolated strain could not be identified. The C. trachomatis which causes inclusion conjunctivitis in Japan appears to have a distribution of serovars similar to that of the sexually transmitted diseases.

Purpose: A human biovar of Chlamydia trachomatis was used to develop a murine model of ocular chlamydial infection. The inbred mouse model will allow detailed immunologic studies during ocular infection, and use of a human biovar for infection may aid in identification of appropriate vaccine strategies against chlamydial infections., Methods: BALB/c, C3H/HeN, and C57B1/6J mice (n = 5 to 10 mice/group) were topically infected in the conjunctiva with C serovar of C. trachomatis. The effects were tested of single and repeated infection with 5000 inclusion-forming units (IFU) in 5 microliters and different inoculum doses. Conjunctival surfaces of both eyes were swabbed for microbiologic signs (isolation culture or direct fluorescent antibody staining) of infection over 4 to 6 weeks. Conjunctivae were removed for histopathologic study, and lymphocytes from draining cervical lymph nodes and spleens were tested for chlamydia-specific proliferative responses. Serum was obtained from all mice and tested for anti-chlamydial antibodies., Results: BALB/c and C3H/HeN mice developed dose-dependent microbiologic, histopathologic, and immunologic evidence of ocular infection. Eyes of mice were culture-positive from day 7 through at least day 21, with the peak of infection at days 10 to 14 after infection. Histopathologically, the development of conjunctival subepithelial mononuclear infiltration, exudate, and loss of goblet cells occurred within 1 week. Dose-dependent lymphoproliferative responses to whole chlamydial elementary bodies were observed; anti-chlamydial antibody was detected by immunoblotting only in infected mice., Conclusions: Several strains of inbred mice are susceptible to human chlamydial biovars and may provide a useful alternative disease model in which to study the immunopathogenesis of ocular chlamydial infection and test of vaccine candidates derived from clinically relevant human biovars.

Purpose: The Kodak Surecell Chlamydia test, a rapid enzyme immunoassay, has been reported to be highly sensitive (93%) and specific (96%) for detecting chlamydial lipopolysaccharide antigen in conjunctival specimens from infants, but has not been evaluated previously in adult conjunctival specimens. This study was designed to determine the efficacy of the Kodak Surecell Chlamydia test for the laboratory diagnosis of adult inclusion conjunctivitis., Methods: Twenty Chlamydia culture-positive conjunctival specimens from adults (true-positives) and 20 true-negative specimens were tested with the Kodak Surecell Chlamydia test., Results: The Kodak Surecell Chlamydia test was 40% (8/20) sensitive, 100% (20/20) specific, and 70% (28/40) efficient., Conclusions: This study indicates that the Kodak Surecell Chlamydia test, though highly specific, is less sensitive in its ability to diagnose chlamydial conjunctivitis in adults than has been reported previously in infants.

Purpose/methods: After we studied a case of chlamydial and adenoviral coinfection in a 20-year-old woman, we determined the incidence of chlamydial infection in patients with acute adenoviral conjunctivitis. In a randomized retrospective study, we evaluated 100 specimens of patients with culture-proven adenoviral conjunctivitis., Results/conclusions: Three of 100 (3%) specimens tested positive for chlamydial DNA using polymerase chain reaction. Adenoviral and chlamydial coinfection is rare, yet should be considered in patients with prolonged follicular keratoconjunctivitis.

Objectives: To determine the prevalence of Chlamydia trachomatis in acute conjunctivitis (non-trachoma) in Australia and to examine the source of transmission., Design: A prospective survey of 400 consecutive patients presenting with acute conjunctivitis to the Royal Victorian Eye and Ear Hospital Emergency Department, Melbourne, from May to November 1991. Patients identified with chlamydial conjunctivitis during the survey period and in the following two months were assessed for concomitant genital infection., Results: Chlamydia was the causative organism in 2% of patients with acute conjunctivitis. Of 15 patients with chlamydial conjunctivitis, 11 presented with disease in one eye only, and the same number had had symptoms for longer than two weeks. Many had been seen previously by experienced ophthalmologists, yet there were long delays in making a definitive diagnosis. Ten of the 12 adult patients who were assessed had signs of concomitant genital tract infection, although none had past or current genital tract symptoms. Serotyping of chlamydial isolates from the genital tract and eye showed concordance in individual patients., Conclusion: Most cases of ocular chlamydia infection have a genital source. Therefore, it is essential that all patients with chlamydial conjunctivitis and their sexual partners are examined and treated for concomitant genital infection.

Purpose: To turn attention to the etiological importance of chlamydia in chronic conjunctivitis., Methods: From December 1993 to November 1994 the authors examined 95 patients with chronic conjunctivitis for the presence of Chlamydia trachomatis antigens in conjunctival epithelial cells using direct immunofluorescent antibody tests., Results: The test was positive in 23 patients and most of them responded positively to general tetracycline treatment combined with locally used Sulfacetamide or Floxal., Conclusion: Chlamydia trachomatis are common causes of conjunctivitis in adults.