Detection Of Chlamydia Trachomatis Research Articles

Detection of Neisseria gonorrhoeae and Chlamydia trachomatis from pooled rectal, pharyngeal and urine specimens in men who have sex with men

ObjectivesScreening of men who have sex with men (MSM) for Chlamydia trachomatis (CT) and Neisseria gonorrhoeae (NG) requires sampling from anorectal and pharyngeal sites in addition to urogenital sampling. Due...

Lymphogranuloma Venereum: a Croatian Case Report of a Patient with Proctitis and Concurrent Cervical Lymphadenitis

Background Lymphogranuloma venereum (LGV) can present with genital ulcers, proctitis, or femoral/inguinal lymphadenopathy with or without constitutional symptoms. There have been reports on the infection characterized by supraclavicular or cervical lymphadenopathy, although there is no published data on the detection of Chlamydia trachomatis (CT) in the affected lymph node.Methods We report a patient with pharyngitis, proctitis and cervical lymphadenitis, in whom LGV specific DNA was detected by polimerase chain reaction (PCR) in both the rectal swab and cervical lymph node fine needle aspirate.ResultsA 48-year-old Croatian HIV-positive male patient attended our outpatient HIV clinic complaining of a 10-day perianal pain, bloody rectal discharge with normal stool consistency and painful and enlarged cervical lymph node. On the first day he had fever, which subsided on the following day. Clinical examination demonstrated exudate on the right tonsil, painful, and enlarged right cervical lymph node (5 × 2 cm, Figure 1), perianal pain on palpation and rectal purulent discharge. During the preceding 10-year period, the patient was taking his antiretroviral therapy regularly and had an undetectable HIV RNA by PCR. We started treatment with ceftriaxone and doxycycline. The fine needle aspirate of the affected lymph node showed a pyogranulomatous inflammation. Indirect immunofluorescence assay on CT antibodies detected positive titers in IgG (>512) and IgA (256) classes. LGV-specific DNA was detected in both the rectal swab and the cervical lymph node fine needle aspirate. Ceftriaxone was discontinued after 5 days and doxycycline therapy was continued for a total of 6 weeks because of the persistence of the cervical lymphadenopathy. The patient came to a full recovery.Conclusion We found six published case reports on LGV with associated cervical lymphadenopathy in which the infection with CT was established by serologic testing and/or by nucleic acid amplification tests of the pharyngeal and/or rectal swabs. Some of the mentioned reports demonstrated granulomatous inflammation present in the affected cervical lymph node, as was evidenced in our patient. To our knowledge, this is the first case report of a patient with CT infection in whom LGV-specific DNA was detected in the cervical lymph node.Disclosures All authors: No reported disclosures.

The Detection Rates of Urogenital Chlamydia trachomatis and Neisseria gonorrhoeae in Chinese Population Applying for U.S. Immigration

Background Medical screening for gonorrhea is under the mandatory requirements of the Center for Disease Control and Prevention for aliens applying for U.S. immigration. Gonorrhea is frequently associated with Chlamydia trachomatis (CT) infection. There is limited data on the detection rates of urogenital CT and Neisseria gonorrhoeae (NG) in Chinese population.Methods Data on physical examinations of applicants in Guangdong International Travel Health Care Center, China were collected and retrospectively analyzed. The nucleic acids of urogenital CT and NG from urine specimens were detected by fluorescent probe PCR using cobas 4800 CT/NG Amplication Detection Kit (Roche Molecuar Systems, Inc.). The detection rates of CT and NG among the overall population were assessed. In addition, the detection rates of CT by age and gender were also evaluated.ResultsIn total, 10,549 applicants underwent physical examinations from September 2016 to March 2017. Mean (SD) age was 41.4 (15.6) years, ranging from 15 to 90 years. The proportion of females (56.17%) was higher than that of males (43.83%). Of the 10,512 people who completed the detection of CT, 369 (3.51%) were CT-positive. The detection rate of CT was significantly higher in females (4.15%) than males (2.69%) (Table 1). The highest CT-positive rate was observed in the age group of 25–29 years (24/344, 6.98%) among males, while in a slightly younger age group of 20–24 years (34/524, 6.49%) among females (Figure 1). Of the 10,510 applicants who completed the detection of NG, 10 (0.10%) were NG positive. The detection rate of NG in males and females was 0.11 and 0.08%, respectively (Table 1).Conclusion Our study first reported the detection rates of urogenital CT (3.51%) and NG (0.10%) among Chinese applicants for U.S. immigration. The highest positive rate of CT was observed among young and middle-aged people, which should gain more attention.Table 1:The positive rate of the detection of CT and NGItemGenderPositive numberPositive rate (95% CI) (%)CTMale (n = 4606)1242.69 (2.22–3.16)Female (n = 5906)2454.15 (3.64–4.66)Total (n = 10,512)3693.51 (3.16–3.86)NGMale (n = 4606)50.11 (0.01–0.20)Female (n = 5904)50.08 (0.01–0.16)Total (n = 10,510)100.10 (0.04–0.15)CT: chlamydia trachomatis; NG: neisseria gonorrhoeae; CI: confidence interval.Disclosures All authors: No reported disclosures.

Corrigendum: Rapid and visual Chlamydia trachomatis detection using loop-mediated isothermal amplification and hydroxynaphthol blue.

Corrigendum: Rapid and visual Chlamydia trachomatis detection using loop-mediated isothermal amplification and hydroxynaphthol blue.

A simple and rapid DNA extraction method for Chlamydia trachomatis detection from urogenital swabs

A highly sensitive and specific Chlamydia trachomatis (CT) diagnostic test was developed by combining filtration isolation of nucleic acid (FINA) extraction with quantitative polymerase chain reaction including an internal control to identify test inhibition. A pilot study of 40 clinical specimens yielded 100% sensitivity and specificity.

Prevalence of genital Chlamydia trachomatis infections in Russia: systematic literature review and multicenter study.

A reliable overview of data on the prevalence of Chlamydia trachomatis (CT) in Russia is lacking and needed. All the available data on CT prevalence were analyzed in a systematic literature review on CT prevalence in Russia, strengthened with data from the multicenter study among 1263 people in the second-largest Russian megalopolis, St. Petersburg, testing for CT DNA in urethral, anal, cervical and prostate samples. A total of 10 articles met the inclusion criteria. The overall average prevalence of genital CT infections in Russian populations ranged from 2.9% to 33%. Risk factors included being symptomatic (P = 0.004; in men P < 0.001), being younger than 30 years (P = 0.001) and being a man who has sex with men (MSM) (P = 0.0084). Main limitations included the lack of studies in MSM. CT prevalence was higher in the groups where urethral and prostate secretion samples were pooled (5.2%-7.3% vs 3.2% in the urethra only). The data on CT prevalence in a range of Russian populations are analyzed and reported. Prostate secretions represent an additional sampling material for the study of CT infection in men. CT detection in some settings in St. Petersburg yielded levels of reliability comparable with internationally available tests. The initiation of screening programs for Chlamydia infections in Russia should be considered.

Sensitivity and Specificity of Empiric Treatment for Sexually Transmitted Infections in a Pediatric Emergency Department

To determine test characteristics of provider judgment for empiric antibiotic provision to patients undergoing testing for a sexually transmitted infection. We conducted a retrospective cross-sectional electronic health record review of all patients aged 13-19 years who had Neisseria gonorrhoeae (GC) and Chlamydia trachomatis (CT) testing sent from an urban, academic pediatric emergency department in 2012. We abstracted data, including patient demographics, chief complaint, sexually transmitted infection test results, and treatment. We calculated test characteristics comparing clinician judgment for presumptive treatment for a sexually transmitted infection with the reference standard of the actual results of testing for a sexually transmitted infection. Of 1223 patient visits meeting inclusion criteria, 284 (23.2%) had a positive GC and/or CT test result. Empiric treatment was provided in 615 encounters (50.3%). Provider judgment for presumptive treatment had an overall sensitivity of 67.6% (95% CI, 61.8-73.0) and a specificity of 55% (95% CI, 51.7-58.2) for accurate GC and/or CT detection. Many adolescents tested for GC and CT receive empiric treatment at the initial emergency department visit. Provider judgment may lack sufficient sensitivity and specificity for identifying infected patients, resulting in the potential for undertreatment of true disease, overtreatment of uninfected patients, or both.

Evaluation of self-collected rectal swabs for the detection of bacteria responsible for sexually transmitted infections in a cohort of HIV-1-infected patients.

Purpose. The standard approach to screening sexually transmitted infections (STIs) has often been restricted to urogenital specimens. Most current guidelines, however, also recommend testing extra-genital sites, including rectal locations, because asymptomatic rectal carriage of pathogens has often been reported. The aim of our study was to evaluate self-collected rectal swabs to screen bacterial STIs in HIV-infected patients in Marseille, France.Methodology. Between January 2014 and December 2015, 118 HIV-infected patients (93 males and 25 females) agreed to self-sample anal swabs for detection of bacterial STI. Detection of Neisseria gonorrhoeae, Chlamydia trachomatis, Treponema pallidum, Mycoplasma genitalium and Haemophilus ducreyi was performed using in-house qPCR assay.Results/Key findings. Bacterial STIs were found in 8 % (9/118) of the patients. C. trachomatis was the most commonly detected bacterium (4.2 %) followed by N. gonorrhoeae (2.5 %), M. genitalium (1.7 %) and T. pallidum (0.8 %). All the positive patients were males. The rectal carriage of pathogenic bacteria was fortuitously discovered for seven men (78 %) who did not present rectal signs of STIs and was suspected for two men who presented proctitis (22 %).Conclusion. In conclusion, testing extra-genital sites is crucial for the diagnosis of STIs in men and women presenting or not concomitant urogenital infections in order to detect asymptomatic carriage with the aim of controlling and preventing transmission to their sexual partners.

Rapid detection and strain typing of Chlamydia trachomatis using a highly multiplexed microfluidic PCR assay.

Nucleic acid amplification tests (NAATs) are recommended by the CDC for detection of Chlamydia trachomatis (Ct) urogenital infections. Current commercial NAATs require technical expertise and sophisticated laboratory infrastructure, are time-consuming and expensive, and do not differentiate the lymphogranuloma venereum (LGV) strains that require a longer duration of treatment than non-LGV strains. The multiplexed microfluidic PCR-based assay presented in this work simultaneously interrogates 13 loci to detect Ct and identify LGV and non-LGV strain-types. Based on amplified fragment length polymorphisms, the assay differentiates LGV, ocular, urogenital, and proctocolitis clades, and also serovars L1, L2, and L3 within the LGV group. The assay was evaluated in a blinded fashion using 95 clinical swabs, with 76 previously reported as urogenital Ct-positive samples and typed by ompA genotyping and/or Multi-Locus Sequence Typing. Results of the 13-plex assay showed that 51 samples fell within urogenital clade 2 or 4, 24 samples showed both clade 2 and 4 signatures, indicating possible mixed infection, gene rearrangement, or inter-clade recombination, and one sample was a noninvasive trachoma biovar (either a clade 3 or 4). The remaining 19 blinded samples were correctly identified as LGV clade 1 (3), ocular clade 3 (4), or as negatives (12). To date, no NAAT assay can provide a point-of-care applicable turnaround time for Ct detection while identifying clinically significant Ct strain types to inform appropriate treatment. Coupled with rapid DNA processing of clinical swabs (approximately 60 minutes from swab-in to result-out), the assay has significant potential as a rapid POC diagnostic for Ct infections.

Evaluation of a dilution method for non-evaluable results in the detection of Chlamydia trachomatis and Neisseria gonorrhoeae with the Cobas 4800 platform

IntroductionA variable percentage of samples analysed using the Cobas 4800 assay can give an invalid result by PCR inhibition or erroneous due to incorrect DNA extraction with the Cobas 4800 CT/NG test. MethodAn analysis was performed using the vortex agitation and dilution protocol on the original sample (swab or urine) for a total of 116 samples. In order to analyse the sensitivity of this method, 100 samples (swabs and urine) with known results were retested. ResultsA total of 98.3% (114/116) of the samples analysed were resolved with this protocol with 100% agreement after reviewing clinical data, Gram stain, and other samples analysed in parallel from the same patient. DiscussionThe data indicate no loss of sensitivity with this protocol; thus Cobas 4800 users could use this method without the need for alternative methods.

Molecular detection of Chlamydia trachomatis and semen quality of sexual partners of infertile women.

Chlamydia trachomatis is considered as the bacterium that is more sexually transmitted as cause of male urethritis, epididymitis, orchitis and infertility. A total of 116 semen samples of men whose couples are infertile women were analysed. The quality of the semen was measured by standard procedures recommended by WHO while C. trachomatis was detected by the PCR assay. Thirty-seven semen samples were positive for C. trachomatis (31.9%). Regarding semen analysis, no different values were observed between positive and negative samples to C. trachomatis. However, the presence of leucocytes and erythrocytes suggests an inflammatory process; however, these were high in negative samples to C. trachomatis. Furthermore, an association between low seminal volume at 1, 5ml and the positivity to C. trachomatis was observed (OR=2, 1; CI95 % 1,16-3,07). The total semen volume is a contribution by the various accessory glands (this reflects the secretory activity of the glands); a low semen volume could be due to an obstruction of the ejaculatory duct or infection of accessory glands by C. trachomatis. More studies are necessary to identify the causes of a reduced semen volume.

Prevalencia de infecciones de transmisión sexual en pacientes sintomáticos y asintomáticos de Yucatán

IntroductionEpidemiological data about non-viral sexually transmitted infections (STI) in Yucatan are limited and come from serological methods. Material and methodsRetrospective study to estimate the prevalence of STI in symptomatic and asymptomatic patients from Yucatan based on a molecular method. The urogenital samples were taken from 147 patients (53 men and 94 women) to extracted nucleic acids. Real-time polymerase chain reaction was used to simultaneous detection of Chlamydia trachomatis, Neisseria gonorrhoeae, Mycoplasma genitalium, Mycoplasma hominis, Ureaplasma urealyticum, Ureaplasma parvum y Trichomonas vaginalis. Statistical analysis was performed with the Fisher's exact test. ResultsThe prevalence of STIs was 45.6%. Ureaplasma spp. was the pathogen most frequent and the females the most affected (p=.022). It was detected 12.2% of coinfections being more frequent in women (16.0 vs. 3.7%, p=.035). No significant differences were found between symptomatic patients (n=138) with positive polymerase chain reaction (n=64) and negative (n=74). Women 21-40 years were the most exposed to STIs (p<.05). ConclusionsThese data confirm the high prevalence of non-viral STIs in Yucatán, being the first epidemiological report based on a molecular method.

Development, validation and testing costs of an in-house real-time PCR assay for the detection of Chlamydia trachomatis.

To improve the screening of Chlamydia trachomatis(C. trachomatis) in Brazil, an accurate and affordable method is needed. The objective of this study was to develop and assess the performance and costs of a new in-house real-time PCR (qPCR) assay for the diagnosis of C. trachomatis infection. Asymptomatic women aged 14-25 years who attended primary health services in Manaus, Brazil, were screened for C. trachomatis using the Digene Hybrid Capture II CT-ID (HCII CT-ID) DNA test. A subset of cervical specimens were tested using an in-house qPCR and a commercial qPCR, ArtusC. trachomatis Plus RG PCR 96 CE (Artus qPCR) kit, as a reference test. A primer/probe based on the sequence of cryptic plasmid (CP) was designed. An economic evaluation was conducted from the provider's perspective. The primers were considered specific for C. trachomatis because they did not amplify any product from non-sexually transmitted bacterial species tested. Overall, 292 specimens were tested by both the commercial kit (Artus qPCR) and the in-house qPCR. Of those, one resulted in no amplification and was excluded from the analysis. The sensitivity, specificity, and positive and negative predictive values of the in-house qPCR were 99.5 % [95 % confidence interval (CI): 97.1-100], 95.1 % (95 % CI: 89-98.4), 97.4 % (95 % CI: 94-99.1) and 99.0 % (95 % CI: 94.5-100), respectively. The cost per case of C. trachomatis was £0.44 ($0.55) for HCII CT-ID, £1.16 ($1.45) for Artus qPCR and £1.06 ($1.33) for in-house qPCR. We have standardized an in-house qPCR to detect cervical C. trachomatis targeting CP. The in-house qPCR showed excellent accuracy and was more affordable than the commercial qPCR kit.

Development a rapid and accurate multiplex real time PCR method for the detection Chlamydia trachomatis and Mycoplasma hominis.

Sexually transmitted diseases easily spread among sexually active people and often have no symptoms. Rapid and accurate method for detecting these infections are necessary in early stages. The traditional detection methods of them are difficult and time-consuming. In this study, multiplex real time PCR was optimized for rapid identification of Chlamydia trachomatis and Mycoplasma hominis in a single tube and was performed with our designed primers. The sensitivity test was carried out to designed primers with diluted genomic DNA. To defined the specificity, non STD bacteria were used as DNA template. This study indicated that the developed multiplex real time PCR can be an effective alternative procedure to the conventional methods for rapid and accurate identification of C Chlamydia trachomatis and Mycoplasma hominis. Multiplex real-time PCR Results of them were checked with melting curves. The sensitivity of our designed primer by multiplex real time PCR for Chlamydia trachomatis and Mycoplasma hominis were 4.78×1010 and 8.35×1010 , respectively, Which the primers did not amplify any product from a non-STD species. Multiplex real time PCR by our new primers and analysis of melting curves were successfully usable for rapid and accurate detection of Chlamydia trachomatis and Mycoplasma hominis. This assay instead of traditional culture method, has considerable potential to be rapid, accurate and highly sensitive molecular diagnostic tool for simultaneous and direct detection.

The new frontier of diagnostics: Molecular assays and their role in infection prevention and control.

Recent advances in technology over the last decade have propelled the microbiology laboratory into a pivotal role in infection prevention and control. The rapid adaptation of molecular technologies to the field of clinical microbiology now greatly influences infectious disease management and significantly impacts infection control practices. This review discusses recent developments in molecular techniques in the diagnosis of infectious diseases. It describes the basic concepts of molecular assays, discusses their advantages and limitations, and characterizes currently available commercial assays with respect to cost, interpretive requirements, and clinical utility.

Evaluation of Cortez OneStep Chlamydia Rapicard™ Insta Test for the Detection of Chlamydia trachomatis in Pregnant Women at Mbare Polyclinic in Harare, Zimbabwe.

Background:Cervical chlamydia infection poses high risk of pregnancy complications and neonatal infection. Reference methods for the detection of chlamydia infection are not available for routine use in developing countries. Point-of-care (POC) tests can bridge this gap. This study evaluated Cortez Onestep Chlamydia Rapicard™ insta test for the detection of Chlamydia trachomatis in pregnant women at Mbare Polyclinic and determined the prevalence of C. trachomatis.Methods:This was a cross sectional study in 242 pregnant women aged ≥18 years attending their first ANC visit at Mbare polyclinic in Harare, Zimbabwe. Data collection form was used to obtain demographic and predisposing factors to Chlamydia infection and two endocervical swabs were collected from each patient. One specimen was examined by the POC test at the clinic and the other by SDA method in the laboratory.Results:The sensitivity, specificity, positive and negative predictive values of the rapid kit were 71.4%, 99.6%, 90.9% and 98.3% respectively. Prevalence of C. trachomitis was 5.8% by SDA method.Conclusion and Global Health Implications:The kit’s sensitivity (71.4%) and specificity (99.6%) implies that the rapid test is an important test which needs further evaluations. The prevalence of C. trichomitis of 5.8% is comparable to studies done elsewhere in Africa.

Non-viral sexually transmitted infections - Epidemiology, clinical manifestations, diagnostics and therapy : Part1: Gonococci

Approximately 1 million people are infected per day worldwide by one or more sexually transmitted infections (STI) as estimated by the World Health Organization (WHO). Gonorrhoea represents an almost exclusively sexually transmitted infection, which predominantly affects mucous membranes of the genitourinary tract. Extragenital localization of infections is also possible, e. g. in the anorectal region. Currently, only syphilis and human immunodeficiency virus (HIV) are notifiable diseases according to the Infection Protection Act in Germany. In Saxony, an extended registration ordinance according to the German Infection Protection Act is in force, which means that besides syphilis the laboratory detection of Neisseria gonorrhoeae, Chlamydia trachomatis and genital mycoplasms are also notifiable infections. In particular, beginning in 2009 in Saxony aspectacular increase of registered infections due to N. gonorrhoeae was observed and in 2015 altogether 824 infections due to N. gonorrhoeae were reported. Alarming is the increase in resistance of N. gonorrhoeae against penicillin, doxycycline, ciprofloxacin and recently also against azithromycin and third generation cephalosporins. The so-called superbug of N. gonorrhoeae, which originated in Japan with multidrug resistance against most of the currently available oral antibiotics, has now arrived in Europe. Intramuscular or intravenous injection of ceftriaxone plus oral azithromycin, each given as single dose is the standard therapy for gonorrhoea.

Rapid and visual Chlamydia trachomatis detection using loop-mediated isothermal amplification and hydroxynaphthol blue.

Chlamydia trachomatis is the major bacterial sexually transmitted disease worldwide. The clinical symptoms are broad, and chronic C.trachomatis infections could lead to blindness, ectopic pregnancy, sterility in males and females, and a higher risk of the development of cervical cancer. The result indicates the potential usefulness of our crude DNA lysis coupled loop-mediated isothermal amplification with hydroxynaphthol blue for a simple, rapid, specific, sensitive and cost-effective assay for C.trachomatis detection from suspected specimens. This assay offers an alternative in the clinical diagnosis of C.trachomatis in resource-limited health-care facilities and clinical laboratories in developing countries, and in field tests.

Validación técnica de una PCR: Reacción en cadena de la polimerasa para la detección de Chlamydia trachomatis

Validación técnica de una PCR: Reacción en cadena de la polimerasa para la detección de Chlamydia trachomatis

Mycoplasma genitalium and Trichomonas vaginalis in France: a point prevalence study in people screened for sexually transmitted diseases

ObjectiveMycoplasma genitalium and Trichomonas vaginalis are common causes of sexually transmitted infections, but limited prevalence data are available in France. We aimed to evaluate the prevalence of M. genitalium and T. vaginalis infections and to assess prevalence by gender, age, sample collection sites and clinical symptoms. A multicentre collection of specimens was intended to obtain a nationwide overview of the epidemiology. MethodsBetween September 2014 and January 2015, a total of 2652 consecutive urogenital specimens submitted to the microbiology diagnostic departments of 16 French university hospitals for Chlamydia trachomatis and Neisseria gonorrhoeae detection were collected. M. genitalium and T. vaginalis prevalence were evaluated using a commercial real-time PCR kit. Clinical data from patients were anonymously collected. ResultsT. vaginalis and M. genitalium prevalence were 1.7% (95% confidence interval 1.3–2.4) and 3.4% (95% confidence interval 2.8–4.2), respectively, and did not differ between gender or age groups, except M. genitalium prevalence between men and women in the 35- to 44-year age group (5.9 vs. 1.5%; p 0.03). M. genitalium prevalence was significantly higher in patients receiving care in sexually transmitted infection clinics, abortion centres, family planning clinics and prisons than in gynaecologic, obstetric and reproduction centres (4.0 vs. 1.7%, p 0.009). Among M. genitalium– and T. vaginalis–positive patients, 70.9 and 61.5% were asymptomatic, respectively. ConclusionsThe low T. vaginalis prevalence does not justify systematic screening for this organism in France. Conversely, selective screening for M. genitalium may be warranted in care settings that receive presumably high-risk sexual behaviour patients, regardless of symptoms.