Diagnosis of a neonatal ophthalmic discharge, Ophthalmia neonatorum, in the molecular age: investigation for a correct therapy.

An early double case of acute Ophthalmia neonatorum in 3-day-old twins is reported. Culture of eye swabs showed a wide bacterial polymorphism, in which common bacteria, such as Klebsiella pneumoniae, Streptococcus pneumoniae, Corynebacterium ulcerans and other Enterobacteriaceae, coexisted with atypical Mycoplasmataceae and Chlamydiaceae from resident cervical-vaginal maternal microbiota. The neonates were in an apparently healthy state, but showed red eyes with abundant greenish-yellow secretion, mild chemosis and lid edema. The maternal cervical-vaginal ecosystem resulted differently positive to the same common cultivable, atypical bacteria culturally and molecularly determined. This suggested a direct maternal-foetal transmission or a further foetal contamination before birth. An extended culture analysis for common bacteria to atypical ones was decisive to describe the involvement of Mycoplasmas (M. hominis and U. urealyticum) within the scenario of the Ophthalmia neonatorum in a Caucasian couple. The introduction of a routine PCR molecular analysis for Chlamydiaceae and N. gonorrhoeae allowed to establish which of these were present at birth, and contributed to determine the correct laboratory diagnosis and to define an adequate therapeutic protocol obtaining a complete resolution after one year for culture and atypical bacteria controls. This study suggests to improve the quality of laboratory diagnosis as unavoidable support to a correct clinical diagnosis and therapy, in a standardized modality both for swabbing and scraping, to check the new-born microbial programming starting in uterus, overtaking the cultural age to the molecular age, and to revise the WHO guidelines of SAFE Strategy for trachoma eye disease, transforming it into SAFES Strategy where the S letter is the acronym of Sexual ecosystem and behavioural valuation/education.

[Infertility in women: latest developments]. / Infertilität bei Frauen : Neueste Entwicklungen.

BACKGROUND: The incidence of female infertility has not changed since the early 1990s. Based on new data from basic research on infertility, novel options in the diagnostics and treatment of infertility have emerged, besides in vitro fertilization (IVF) and intracytoplasmic sperm injection (ICSI). AIM: This review summarizes the current knowledge on female infertility and on modern concepts in diagnostics and treatment. METHODS: A literature research on the causes of infertility and on treatment options was performed, including demographic factors, infectiology, anatomy, endocrinology and metabolism, endometriosis, lifestyle and environmental factors, and psychological factors. RESULTS: Chlamydial infection is still the major cause of tubal infertility. Improvement of the patient's fertility by correction of endocrine and metabolic disorders, in particular thyroid dysfunction and glucose metabolism, as well as fertility surgery are of main interest. CONCLUSIONS: Besides assisted reproductive techniques, concepts to optimize individual fertility have gained increasing importance.

Clinic-based testing for rectal and pharyngeal Neisseria gonorrhoeae and Chlamydia trachomatis infections by community-based organizations--five cities, United States, 2007.

CDC recommends screening of at-risk men who have sex with men (MSM) at least annually for urethral and rectal gonorrhea and chlamydia, and for pharyngeal gonorrhea. Although the standard method for diagnosis is culture, nucleic acid amplification (NAA) testing is generally more sensitive and favored by most experts. NAA tests have not been cleared by the Food and Drug Administration (FDA) for the diagnosis of extragenital chlamydia or gonorrhea and may not be marketed for that purpose. However, under U.S. law, laboratories may offer NAA testing for diagnosis of extragenital chlamydia or gonorrhea after internal validation of the method by a verification study. To determine sexually transmitted disease (STD) testing practices among community-based organizations serving MSM, CDC and the San Francisco Department of Public Health gathered data on rectal and pharyngeal gonorrhea and chlamydia testing at screening sites managed by six gay-focused community-based organizations in five U.S. cities during 2007. This report summarizes the results of the study, which found that three organizations collected samples for NAA testing and three for culture. In total, approximately 30,000 tests were performed; 5.4% of rectal gonorrhea, 8.9% of rectal chlamydia, 5.3% of pharyngeal gonorrhea, and 1.6% of pharyngeal chlamydia tests were positive. These results demonstrate that gay-focused community-based organizations can detect large numbers of gonorrhea and chlamydia cases and might reach MSM not being tested elsewhere. Public health officials could consider providing support to certain community-based organizations to facilitate testing and treatment of gonorrhea and chlamydia.

Association between pubic hair grooming and prevalent sexually transmitted infection among female university students.

Recent findings have suggested an association between pubic hair grooming and self-reported history of sexually transmitted infection (STI), specifically gonococcal infection (GC), chlamydial infection (CT), or human immunodeficiency virus (HIV). We evaluated the association between self-reported extreme grooming and laboratory-confirmed prevalence of GC/CT. Between April 2017 and April 2018, we enrolled English-speaking, adult, female students at a large, Midwestern university who presented on-campus for STI testing. Participants completed a questionnaire on demographics and sexual and grooming behaviors, which was linked to their GC/CT test results based on nucleic acid amplification testing. We defined extreme grooming as removal of all pubic hair either at least weekly in the past 12 months or ≥6 times in the past 30 days. We used two separate logistic regression models to determine whether odds of GC/CT varied by extreme groomer status for either time interval. In the study sample of 214 women, prevalence of GC/CT was 9.8%. Nearly all participants (98.1%) reported ever grooming; 53.6% were extreme groomers in the past year and 18% in the past month. Extreme grooming was not associated with prevalent GC/CT in the past year (odds ratio [OR] = 0.8; 95% confidence interval [CI]: 0.3-1.9; adjusted OR = 0.7; 95% CI: 0.3-2.0) or in the past month (OR = 0.5; 95% CI: 0.1-2.0; aOR = 0.4; 95% CI: 0.1-1.9). Pubic hair grooming was common among female university students attending for STI testing. Findings do not support pubic hair grooming as an STI risk factor in this population.

Survey on Chlamydiaceae in cloacal swabs from Swiss turkeys demonstrates absence of Chlamydia psittaci and low occurrence of Chlamydia gallinacean.

In Switzerland, domestic turkey meat is a niche product. Turkeys are fattened on mixed family-based farms scattered across the country, with most providing access to an uncovered outdoor pasture for the birds. Swiss fattening turkeys may therefore get infected with Chlamydiaceae via wild birds or their faeces, potentially shedding these bacteria at a later stage. The aim of the present study was to acquire baseline data about the shedding of Chlamydiaceae in clinically unremarkable Swiss fattening turkeys at slaughter, potentially exposing slaughterhouse workers to infection. In this large-scale study, 1008 cloacal swabs of Swiss turkeys out of 53 flocks from 28 different grow-out farms with uncovered outdoor pasture were collected over the course of 14 months and examined for the occurrence of Chlamydiaceae by a family-specific 23S-rRNA real-time PCR. Positive samples were further analyzed by Chlamydia psittaci (C. psittaci)-specific real-time PCR and the Arraymate DNA Microarray for species identification. All samples were negative for C. psittaci, but seven swabs out of one flock were tested positive for Chlamydia gallinacea (0.7%). Although turkeys with access to pasture may have contact with Chlamydiaceae-harbouring wild birds or their faeces, the infection rate in Swiss turkeys was shown to be low.

Prevalence of Chlamydiaceae and Mollicutes on the genital mucosa and serological findings in dairy cattle.

It was the aim of this project to obtain information on the prevalence of Chlamydiaceae and Mollicutes and their potential importance for reproductive problems in cattle. Cervical or vaginal swabs were taken from 644 animals in 196 farms and blood samples were collected from 375 cattle. Out of the animals, 6.8% had aborted within the last 12 months, 2.6% showed clinical vaginitis and 11.6% clinical endometritis. Chlamydiaceae were detected and identified by PCR followed by restriction fragment length polymorphism (RFLP) analysis. For the detection and identification of Mollicutes cultivation procedures, biochemical differentiation and serological identification were used. Sera were tested for antibodies against Chlamydiaceae and Mycoplasma (M.) bovis by ELISA and against M. bovigenitalium by Western blot analysis. Chlamydophila (Cp.) abortus was found in three cervical swabs. Cp. pecorum was detected in 9% of cervical or vaginal swabs. The majority of Cp. species found was Cp. pecorum and thus fertility problems caused by Cp. abortus are limited. M. bovis was found in only one genital swab. M. bovigenitalium was rarely diagnosed (3% of cervical and 2% of vaginal swabs). M. bovigenitalium was found more often in cattle having aborted (4/32 animals) than in cattle without history of abortion (5/220, p<0.05). Ureaplasma (U.) diversum existed in 12% of cervical and 36% of vaginal swabs and was found in 8 out of 17 animals with vaginitis. Out of the animals tested, 44.9% were seropositive for Chlamydiaceae, 14.8% for M. bovis and 27.3% for M. bovigenitalium.

Introduction of Chlamydia trachomatis screening for young women in Germany.

Screening for genital Chlamydia trachomatis infections for young sexually active women was incorporated into routine medical care of German statutory health insured patients starting in January 2008. The primary goal of this new preventive measure is the reduction of severe sequelae for women such as tubal infertility and ectopic pregnancies. The course of the deliberations leading to the Federal Joint Committee's decision is summarized in this review.

Experiences with the new genetic variant of Chlamydia trachomatis in Orebro county, Sweden - proportion, characteristics and effective diagnostic solution in an emergent situation.

A Chlamydia trachomatis variant that contains a 377 bp deletion in the cryptic plasmid was recently reported in Sweden. This deletion includes the targets for Cobas Amplicor, Cobas TaqMan48, and Abbott m2000. We examined the proportion and characteristics of this variant in Orebro county, Sweden and developed an effective diagnostic solution. In total, 2,401 consecutive C. trachomatis culture samples and 536 PCR samples from symptomatic and asymptomatic patients and screened females were included. Culture, Cobas Amplicor, and LightMix 480HT were used for diagnosis. A mutant-specific PCR, plasmid sequencing, omp1 sequencing and multilocus sequence typing (MLST) were used to identify and characterise mutants. In total, 162 (6.7%) of the cultured samples were positive for C. trachomatis. However, 61 (38%) of those were negative when using Cobas Amplicor, and 60 of these were subsequently confirmed as the new variant. 13 of these mutant isolates were further characterised genetically, and all were of identical genotype E and the unique MLST sequence type: 21, 19, 1, 2, 1. Of all culture-positive samples, 161 of 162 were positive in the LightMix 480HT assay. The single negative sample was only weakly positive in culture, and negative in all PCRs. Of the 536 PCR samples, 37 were positive in both Cobas Amplicor and LightMix 480HT, 13 were only positive in LightMix 480HT (mutants), and two were only positive in Cobas Amplicor. Mutated C. trachomatis were prevalent in Orebro county in the period from October 2006 to February 2007, and it appeared to be a single clone. LightMix 480HT seemed sensitive, specific, and enabled high throughput diagnostics. However, rare low positive samples may be false-negative. Frequent surveillance and evaluations of diagnostic methods worldwide are crucial.

[Value of Western blotting in serodiagnosis of chlamydial infections]. / Pouzití metody Western blot v sérologické diagnostice chlamydiových infekcí.

UNLABELLED: The Western blot (WB) method was verified for serological diagnosis of chlamydial infections. MATERIAL AND METHODS: For testing, sera previously examined by the microimmunofluorescence (MIF) test with either ambiguous results or those suggesting persistent infection were used. RESULTS: Whereas the investigation confirmed adequate sensitivity and specificity of the MIF test for diagnosing Chlamydophila pneumoniae infection, it was less sensitive in case of Chlamydia trachomatis. Long-term persistence of IgA antibodies, detected by the MIF test, was often not confirmed by WB. CONCLUSIONS: The results support the view that detecting antibodies alone, without appropriate clinical symptoms, is not sufficient for antibiotic treatment of any infection.

Survey of Sexually Transmitted Disease Laboratory Methods in U.S. Army Laboratories, 2012.

INTRODUCTION: Sexually transmitted diseases (STD) affect primarily young people (17-24 years). The U.S. Military, with many young people, strives to maintain effective STD treatment and prevention programs using current methods. Laboratory testing technology and capacity are important for appropriate clinical management and to provide data to direct prevention programs. STD laboratory capabilities are assessed in civilian and military laboratories using surveys. An Army laboratory survey was conducted in 2007. The Army laboratory survey reported here was conducted on 2012 to describe STD tests done, laboratory testing practices, and testing volume to include the use of human immunodeficiency virus point-of-care tests and a novel reverse syphilis testing algorithm. MATERIALS AND METHODS: A web-based survey was offered to all 32 Army laboratories in 2013 to assess testing in 2012. Twenty-two laboratories (69%), including all medical center laboratories, completed the survey. The survey was approved by the U.S. Army Human Protection Review Board. RESULTS: The Army laboratories reported testing more than 230,000 specimens for Chlamydia trachomatis (CT) and Neisseria gonorrhoeae (NG), with 82% and 86% using nucleic acid amplification test (NAAT) methods for CT and NG, respectively. Eleven laboratories (50%) performed combined NAAT methods for CT and NG. Four (18%) performed NG antimicrobial susceptibility testing. Two (10%) screened for syphilis using the reverse algorithm. All offered in-house wet-mount microscopy for Trichomonas vaginalis. Thirteen (62%) used rapid human immunodeficiency virus testing. CONCLUSION: Comparing the 2012 results to the 2007 Army survey results, use of NAAT methods remained relatively stable while antimicrobial NG susceptibility testing decreased. Efforts to promote NAAT methods, to include testing vaginal and nongenital specimens for CT and NG, must continue. NG antibiotic resistance testing should be increased. Monitoring the use of the reverse syphilis screening algorithm is recommended to assess the impact of false-positive results.

Microscopic diagnosis of dysbacteriosis in stained vaginal smears in clinical practice.

Dysbacteriosis is a microscopical diagnosis. In women with dysbacteriosis, an overgrowth of coccoid bacteria and almost a complete absence of lactobacilli are observed in the (stained) vaginal smear. The aim of this study was to determine the accuracy of this microscopic diagnosis in clinical practice. The analysis concerned 342 consecutive cases in which the microscopy of the stained smears was performed by general practitioners trained in diagnosing dysbacteriosis. These smears were sent to the pathologist for confirmation of the microscopical diagnosis of the clinician. The cytological diagnoses of the pathologist, sometimes performed on restained slides when the quality of the staining was substandard, were considered as the "gold standard." In 92 of the 342 cases, dysbacteriosis was unequivocally established by the pathologist. Sensitivity and specificity of the microscopical diagnoses of the clinicians were 40% and 85%, respectively. There were 37 false-positive and 54 false-negative diagnoses of dysbacteriosis rendered by the clinicians. The most frequent reason for a false-negative diagnosis was an excess of lactobacilli in the smear. This study shows that even in stained smears it is difficult for clinicians to render a correct evaluation of the status of the vaginal flora.

Clinical characteristics of the afebrile pneumonia associated with Chlamydia trachomatis infection in infants less than 6 months of age.

Respiratory tract colonization with Chlamydia trachomatis commonly occurs in natally acquired chlamydial infection and is sometimes associated with a chronic, afebrile pneumonia that has relatively distinctive clinical characteristics. To further define the frequency and clinical characteristics of lower respiratory tract disease associated with C trachomatis, we grouped 56 infants aged less than 6 months with afebrile pneumonia according to nasopharyngeal shedding of Chlamydia and viruses and compared their illnesses. Forty-one (73%) were positive for C trachomatis (23 had C trachomatis only, while 18 had C trachomatis plus a virus [cytomegalovirus, respiratory synctial virus, adenovirus, rhinovirus, or enterovirus]), and 15 were C trachomatis negative (nine had a virus only, and six had neither C trachomatis nor virus). The 41 infants with C trachomatis alone or C trachomatis plus a virus were similar clinically and differed significantly from other infants in several ways: (1) onset of symptoms before 8 weeks of age; (2) gradually worsening symptoms; (3) presentation for care at 4 to 11 weeks of age; (4) presence of conjunctivitis and ear abnormalities; (5) chest roentgenograms showing bilateral, symmetrical, interstitial infiltrates and hyperexpansion; (6) peripheral blood eosinophils greater than or equal to 300/cu mm; and (7) elevated values for serum immunoglobulins M, G, and A. Pediatrics 63:192--197, 1979, Chlamydia trachomatis, pneumonia, afebrile pneumonia, interstitial pneumonia.

Moving to nucleic acid-based detection of genital Chlamydia trachomatis.

Laboratory diagnosis of the bacterium Chlamydia trachomatis has gone through a complete phase of evolution since it was first identified as a significant cause of sexually transmitted infection. As a fragile, obligatory intracellular organism, it was initially only grown in eggs. Subsequently, diagnosis relied on culture in continuous cell lines. To address the limitations of culture, immunological methods were developed and direct antigen detection using enzyme immunoassay and immunofluorescence flourished. With the advent of molecular technologies, nucleic acid-based amplification techniques became the methods of choice, offering improved standard of care for diagnosis and opening up the possibility of screening using noninvasive, patient-acceptable specimens. In this article, the various currently available molecular methods are examined, some of the existing problems discussed and a view on what we think might happen in the next 5 years to the technology and requirement in diagnosis and screening is given.

Detection of Chlamydia trachomatis using consecutive endocervical swabs. Prevalence in asymptomatic female adolescents and women attending a sexually transmitted disease clinic.

Chlamydia trachomatis is an organism commonly transmitted through sexual intercourse. In women it is associated with cervicitis, salpingitis, perihepatitis and infertility. Neonates born to infected women may have inclusion conjunctivitis and pneumonia due to this organism. Screening in obstetrics and in gynecology clinics is not usually performed routinely because of the cost and time involved in culturing the organism. A rapid enzyme immunoassay (EIA) kit (Chlamydiazyme, Abbott Laboratories) that can detect C. trachomatis was developed recently. Women belonging to two different cohorts were studied to determine whether multiple endocervical samples increased the likelihood of a positive result from this EIA kit. One cohort consisted of 70 asymptomatic, sexually active female adolescents from a local family planning clinic. The second cohort included 80 women who were seen at a sexually transmitted disease (STD) clinic. Both groups were assayed for Chlamydia infections using the rapid EIA kit. Positive test results were found in 7 of the 70 asymptomatic teenagers (10%) and 12 of the 80 women from the STD clinic (15%). No significant differences were noted in the order of the positive swabs in either group, although more of the earlier swabs tended to be positive.

[Chlamydial diseases of domestic animals--zoonotic potential of the agents and diagnostic issues]. / Chlamydienerkrankungen der Nutz- und Haustiere--Zoonotisches Potential der Erreger und diagnostische Fragen.

The role of chlamydiae as agents of a number of important animal and human diseases is still the subject of intensive research. Recently, a proposal for taxonomic reclassification of this group of obligate intracellular bacteria was published, which was based on a large amount of new data on genetic relatedness. According to this proposal, the family Chlamydiaceae now comprises two genera (Chlamydia and Chlamydophila) with 9 largely host-related species. The previously accepted classification scheme had distinguished 4 species within the genus Chlamydia. The most important animal chlamydiosis with zoonotic character is psittacosis, a systemic disease in psittacine birds of acute, protracted, chronic or subclinical manifestation. The analogous infection in domestic and wild fowl is known as ornithosis. Avian strains of C. psittaci (new classification: Chlamydophila psittaci) can also infect humans, the symptoms being mainly unspecific and influenza-like, but severe pneumonia, endocarditis and encephalitis are also known. The main group of persons facing an elevated risk of infection includes those having frequent contact with domestic and companion birds at work or in their spare time. In Germany, the annual average of notified cases is approximately 100. Cases of transmission to humans were repeatedly reported in connection with enzootic abortion in sheep (causative agent: C. psittaci or Chlamydophila abortus, respectively). Various chlamydial species occur as pathogens and commensals as well in cattle, pigs, horses, and cats. The assessment of the actual epidemiological importance is, however, often difficult because of their almost ubiquitous spread. Likewise, those strains of C. pneumoniae (new classification: Chlamydophila pneumoniae) found in several animal species can not yet be assessed for pathogenic properties. The possibilities for diagnostic detection of chlamydiae have considerably improved following the introduction of molecular methods, particularly the polymerase chain reaction (PCR), which permits direct identification from clinical specimens and differentiation of species.

Identification of Chlamydial species in crocodiles and chickens by PCR-HRM curve analysis.

Recently, a PCR protocol (16SG), targeting 16S rRNA gene coupled with high resolution melt (HRM) curve analysis was developed in our laboratory and shown to reliably detect and identify the seven different Chlamydiaceae spp. In this study, the potential of this method was assessed for detection and differentiation of Chlamydiosis in clinical specimens. Of the total number of 733 specimens from a range of animal species, 219 (30%) were found positive by 16SG PCR. When a sufficient amount of DNA was available (64 submissions), amplicons generated by the 16SG PCR were subjected to HRM curve analysis and results were compared to that of nucleotide sequencing. In all instances, the infecting Chlamydiaceae spp. was genotyped according to the identity of its nucleotide sequence to a reference species. Analysis of the HRM curves and nucleotide sequences from 16SG PCR amplicons also revealed the occurrence of a Chlamydophila-like, a Parachlamydia-like and a variant of Chlamydophila psittaci in chickens. These results reveal the potential of 16SG PCR-HRM curve analysis for rapid and simultaneous detection and identification of Chlamydiaceae spp. in animals and demonstrate the capacity of this system for rapid identification of new Chlamydiaceae spp. in animals during routine diagnostic testings.