Urogenital Swabs Research Articles

Detection of Chlamydia trachomatis and Neisseria gonorrhoeae with the cobas CT/NG v2.0 test: performance compared with the BD ProbeTec CT Qx and GC Qx amplified DNA and Aptima AC2 assays

ObjectivesInfections due to Chlamydia trachomatis (CT) and Neisseria gonorrhoeae (NG) are among the most common bacterial sexually transmitted infections worldwide, most of which are asymptomatic. Detection of infection using a...

Comparison of rRNA-based and DNA-based nucleic acid amplifications for detection of Chlamydia trachomatis, Neisseria gonorrhoeae, and Ureaplasma urealyticum in urogenital swabs

BackgroundNucleic acid amplification tests (NAAT) are well-accepted in diagnosis and surveillance of sexually infectious pathogens worldwide. However, performance differences between a RNA-based NAAT and DNA-based NAAT are rarely reported. This study compares the performances of the RNA-based SAT (simultaneous amplification and testing) assay and the DNA-based quantitative real-time polymerase chain reaction (qPCR) assay.MethodsA total of 123 urogenital swabs were collected from outpatients with suspected genital infections in our hospital. Chlamydia trachomatis (CT), Neisseria gonorrhoeae (NG), and Ureaplasma urealyticum (UU) in these swabs were simultaneously tested by SAT and qPCR. Any swabs were positive in the qPCR assay were further verified by following cloning and sequencing. All statistical analysis was performed using the SPSS software.ResultsWhen the concentrations of CT, NG, or UU were more than 1 × 103 copies/ml, 100% agreements between SAT and qPCR were observed regardless of the pathogen. No discrepancy was found. However, the sensitivity of SAT is significantly higher than qPCR in samples with concentration less than 1 × 103 copies/ml. When tested by SAT and qPCR, 57.14 and 28.57% were positive for CT, 46.15% and 0 were positive for NG, 80% and 0 were positive for UU, respectively.ConclusionsThe SAT assay has better agreements and higher sensitivities when compared with the qPCR assay, and thus could be a better choice for screening, diagnosis, and surveillance of sexually transmitted diseases, especially for CT and NG.

Multiplex TaqMan real-time PCR platform for detection of Neisseria gonorrhoeae with decreased susceptibility to ceftriaxone

A multiplex TaqMan real-time PCR platform was developed in this study for combined detection of opa and/or porA genes (identification of N. gonorrhoeae) and the key mutations (Ala501Val/Thr/Pro, and/or Gly545Ser) in penicillin-binding protein 2 (PBP2) associated with decreased susceptibility to extended-spectrum cephalosporins (ESCs). Firstly, the specificities of the TaqMan probes/primers for the multiplex TaqMan real time PCR platform were confirmed by Basic Local Alignment Search Tool (BLAST) analysis. Then the multiplex PCR platform was performed on 77 isolates with decreased susceptibility to ceftriaxone (CRO) and 100 isolates with full susceptibility to CRO under universal optimized reaction conditions. As a result, based on cultivation-based matrix-assisted laser desorption ionization–time of flight mass spectrometry (MALDI-TOF MS) and antimicrobial susceptibility testing in vitro, the multiplex platform had a sensitivity of 100% and a specificity of 95.0% for identifying cultured isolates of Neisseria gonorrhoeae (N. gonorrhoeae, NG, GC) with decreased susceptibility to CRO. When directly screening N. gonorrhoeae with decreased susceptibility to CRO from clinical urogenital swabs, the multiplex platform offered a sensitivity of 96.1% and a specificity of 95.0%. Therefore, on the basis of sample culture and antimicrobial susceptibility testing in vitro, the multiplex TaqMan real time PCR platform has been proven to be a sensitivity of 100% and a specificity of 95.0% useful tool for screening cultured isolates of N. gonorrhoeae with decreased susceptibility to CRO, which can be finished within 2 days.

RESULTS OF DETERMINING THE SPECIES COMPOSITION OF VAGINAL LACTOBACILLI USING REAL-TIME PCR TESTING AND BACTERIOLOGICAL METHOD

Aim. To compare the species composition of lactobacilli in women of reproductive age using real time PCR on urogenital samples before and after culturing microorganisms in Blood-Serum-Y^ast extract-Agar (BSYA). Materials and methods. Using real time PCR, we have examined the species composition of vaginal lactobacilli in 25 healthy women of reproductive age. Samples of urogenital swabs (endocervical and vaginal swabs) were studied. Species identification has been carried out twice: in the native clinical material and in the samples received after using microorganism culture technique in BSYA. Results. After culturing vaginal microorganisms in BSYA, L. iners predominance was found only in 2 cases (8%). At the same time, when using native clinical material this species predominance was discovered in 10 samples (40%). Conclusion. When culturing lactobacilli in BSYA, the growth of L. iners is stunted compared to other species of vaginal lactobacilli. The use of cultural testing turned out to be ineffective in case of predominance of L. iners, the species associated with an increased risk of vaginal dysbiosis.

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.

Detection of Common Bacterial Causes of STIs among STD Clinic Attendees by Gram Stain and Culture versus the Microarray Nucleic Acid Hybridization Method

Objectives: The purpose of the study was to determine the prevalence and etiology of predominant sexually transmitted infections (STIs) occurring among the STD and Gynecology Clinic attendees and to compare sensitivity and specificity of conventional versus molecular diagnostic methods. Methods: A total of 1,441 individuals were recruited from the STD clinic at the National Center for Communicable Diseases and Gynecology Clinic at the National Center for Oncology between 2014 and 2015. Urogenital tract swab samples were collected and tested for N.gonorrhoeae, C.trachomatis, and T.vaginalis using conventional (Gram stain and culture) and molecular (Microarray Nucleic Acid Hybridization - MNAHM) diagnostic methods. The final infection status of participants was determined using the “either test positive” strategy. Results: Of the total 1,441 participants, 19.7% were N.gonorrhoeae infected, 12.5% were C.trachomatis infected, and 7.8% were T.vaginalis infected. For T. vaginalis, conventional methods and MNAHM had sensitivities of 17.9% and 98.2%, with specificities of 96.6% and 99.6%, respectively. For N.gonorrhoeae, conventional methods had sensitivity and specificity values of 33.3% and 99.1%, and MNAHM had values of 95.1% and 98.8%, respectively. Conclusion: The study found a high prevalence of N.gonorrhoeae and C.trachomatis among STD and Gynecology Clinic attendees. Conventional methods for detection of N.gonorrhoeae and T.vaginalis were significantly less sensitive than molecular diagnostic methods and didn’t detect other STI pathogens.

Reproductive Microbiomes: Using the Microbiome as a Novel Diagnostic Tool for Endometriosis

Background: Endometriosis is a chronic inflammatory disease which results in significant pain and long term reproductive consequences for up to 50% of infertile women. This study was focused to understand how endometriosis altered the uterine and cervical bacterial community. Methods and findings: Urogenital swabs and uterine washes were collected from 19 pre-menopausal women undergoing laparoscopic surgery for pelvic pain, suspected endometriosis (experimental n=10), and women undergoing laparoscopic surgery for benign ovarian/uterine conditions (control n=9). Patients were followed for the next year and repeat cervical swabs were obtained. Bacterial community composition was assessed from these samples using Illumina next generation 16S rRNA amplicon sequencing. Bacterial communities were significantly different between sample sites, the uterus and cervix, and stage III endometriosis resulted in significant alterations in the cervical bacterial community. Both bacterial richness and phylogenetic diversity increased in association with stage III endometriosis. Surgical intervention resulted in a stabilized cervical bacterial community for a short period of time. Conclusion: Bacterial community profiling may provide a useful diagnostic tool for identifying endometriosis in asymptomatic, infertile women in a clinical setting.

Prevalence and genotyping of Chlamydia trachomatis in symptomatic male patients from Istanbul, Turkey.

This study was conducted to determine the prevalence and distribution of urogenital Chlamydia trachomatis genotypes in symptomatic male patients who were referred to the clinics of the Istanbul Faculty of Medicine. Of 419 urogenital swabs, 57 samples (13.6 %) were positive for C. trachomatis. Genotype distribution of C. trachomatis-positive samples identified five genetic variants namely genotype E as the most prevalent (36.4 %), followed by genotype G (23.6 %), H (21.8 %), D (16.4 %) and F (1.8 %). We believe that this is the first study on distribution of genotypes of C. trachomatis genital infections in symptomatic men in Istanbul, Turkey.

A non-invasive tool for assessing pathogen prevalence in koala (Phascolarctos cinereus) populations: detection of Chlamydia pecorum and koala retrovirus (KoRV) DNA in genetic material sourced from scats

Pathogenic diseases may threaten the viability of wild animal populations, especially when already vulnerable. The mitigation of risks associated with pathogenic infections in populations is an important factor in conservation strategies. Koalas are of conservation concern across the north of their range and are affected by two main pathogens; Chlamydia pecorum and the koala retrovirus (KoRV). This study tested whether DNA from C. pecorum and KoRV could be detected in genetic material isolated from koala scats. Detection of C. pecorum in scat isolated DNA samples was compared with results obtained from urogenital swabs collected from the same individuals as part of an independent study. The ability to detect KoRV in scats from both northern and southern regions of the koala’s range was also assessed. There was a high level of concordance (5/6) between the detection of C. pecorum in DNA isolated from scats and urogenital swabs from the same individual. In positive samples, C. pecorum ompA genotypes were identical between DNA from scats and urogenital swabs in two out of three cases. In samples from the south of the koala’s range, KoRV copy number was higher in DNA isolated from scats compared to DNA isolated from ear tissue, potentially indicating the detection of horizontally acquired infections. Our results demonstrate the ability to detect C. pecorum and KoRV in DNA isolated from koala scats. This method will be useful for studying the prevalence, transmission and impact of these pathogens in wild populations which may subsequently inform conservation management strategies.

PREVALENCE AND PATHOLOGIC FEATURES OF CHLAMYDIA PECORUM INFECTIONS IN SOUTH AUSTRALIAN KOALAS (PHASCOLARCTOS CINEREUS).

Chlamydia pecorum infection is highly prevalent in many koala ( Phascolarctos cinereus ) populations in the eastern states of Australia, causing ocular and urogenital tract disease. In contrast, the current prevalence of chlamydiosis in South Australian (SA) koalas is largely unknown, with few reports of clinical cases. We examined 65 SA rescued wild koalas at necropsy and collected ocular and urogenital swabs for the detection of C. pecorum by PCR. We detected C. pecorum in ocular or urogenital swabs from 57 koalas (88%), and 34 koalas were positive at both ocular and urogenital sites. Clinically overt chlamydial disease was present in only 12 (21%) positive koalas. Gross lesions were often externally inapparent as they affected the urogenital tract (n=5), and 24 infected koalas had microscopically evident lesions only. Lesions were predominantly mild and included conjunctivitis, cystitis, and urethritis. Reproductive tract disease was infrequently observed. We detected C. pecorum in 16 (28%) koalas with no evidence of chlamydial disease, suggesting the presence of subclinical carriers in this population. Based on these findings, chlamydiosis has a higher occurrence in SA koala populations than previously thought, but is most often mild and does not always result in overt clinical disease; inapparent and subclinical infections appear common. Further studies of the prevalence in wild-caught SA koalas are needed along with research into the host and bacterial factors that may influence disease outcome in these animals.

Identification of unusual Chlamydia pecorum genotypes in Victorian koalas (Phascolarctos cinereus) and clinical variables associated with infection.

Chlamydia pecorum infection is a threat to the health of free-ranging koalas (Phascolarctos cinereus) in Australia. Utilizing an extensive sample archive we determined the prevalence of C. pecorum in koalas within six regions of Victoria, Australia. The ompA genotypes of the detected C. pecorum were characterized to better understand the epidemiology of this pathogen in Victorian koalas. Despite many studies in northern Australia (i.e. Queensland and New South Wales), prior Chlamydia studies in Victorian koalas are limited. We detected C. pecorum in 125/820 (15 %) urogenital swabs, but in only one ocular swab. Nucleotide sequencing of the molecular marker C. pecorum ompA revealed that the majority (90/114) of C. pecorum samples typed were genotype B. This genotype has not been reported in northern koalas. In general, Chlamydia infection in Victorian koalas is associated with milder clinical signs compared with infection in koalas in northern populations. Although disease pathogenesis is likely to be multifactorial, the high prevalence of genotype B in Victoria may suggest it is less pathogenic. All but three koalas had C. pecorum genotypes unique to southern koala populations (i.e. Victoria and South Australia). These included a novel C. pecorum ompA genotype and two genotypes associated with livestock. Regression analysis determined that significant factors for the presence of C. pecorum infection were sex and geographical location. The presence of 'wet bottom' in males and the presence of reproductive tract pathology in females were significantly associated with C. pecorum infection, suggesting variation in clinical disease manifestations between sexes.

Routes of Hendra Virus Excretion in Naturally-Infected Flying-Foxes: Implications for Viral Transmission and Spillover Risk.

Pteropid bats or flying-foxes (Chiroptera: Pteropodidae) are the natural host of Hendra virus (HeV) which sporadically causes fatal disease in horses and humans in eastern Australia. While there is strong evidence that urine is an important infectious medium that likely drives bat to bat transmission and bat to horse transmission, there is uncertainty about the relative importance of alternative routes of excretion such as nasal and oral secretions, and faeces. Identifying the potential routes of HeV excretion in flying-foxes is important to effectively mitigate equine exposure risk at the bat-horse interface, and in determining transmission rates in host-pathogen models. The aim of this study was to identify the major routes of HeV excretion in naturally infected flying-foxes, and secondarily, to identify between-species variation in excretion prevalence. A total of 2840 flying-foxes from three of the four Australian mainland species (Pteropus alecto, P. poliocephalus and P. scapulatus) were captured and sampled at multiple roost locations in the eastern states of Queensland and New South Wales between 2012 and 2014. A range of biological samples (urine and serum, and urogenital, nasal, oral and rectal swabs) were collected from anaesthetized bats, and tested for HeV RNA using a qRT-PCR assay targeting the M gene. Forty-two P. alecto (n = 1410) had HeV RNA detected in at least one sample, and yielded a total of 78 positive samples, at an overall detection rate of 1.76% across all samples tested in this species (78/4436). The rate of detection, and the amount of viral RNA, was highest in urine samples (>serum, packed haemocytes >faecal >nasal >oral), identifying urine as the most plausible source of infection for flying-foxes and for horses. Detection in a urine sample was more efficient than detection in urogenital swabs, identifying the former as the preferred diagnostic sample. The detection of HeV RNA in serum is consistent with haematogenous spread, and with hypothesised latency and recrudesence in flying-foxes. There were no detections in P. poliocephalus (n = 1168 animals; n = 2958 samples) or P. scapulatus (n = 262 animals; n = 985 samples), suggesting (consistent with other recent studies) that these species are epidemiologically less important than P. alecto in HeV infection dynamics. The study is unprecedented in terms of the individual animal approach, the large sample size, and the use of a molecular assay to directly determine infection status. These features provide a high level of confidence in the veracity of our findings, and a sound basis from which to more precisely target equine risk mitigation strategies.

P05.05 Neisseria gonorrhoeae in indonesia: prevalenceand antimicrobial susceptibility among sti clinics patients in jakarta, yogyakarta and denpasar

Introduction In Indonesia, syndromic management is used for diagnosis and treatment of sexually transmitted infections (STIs), but this approach does not provide accurate data to assess the epidemiology of gonorrhoea and antimicrobial susceptibility of circulating Neisseria gonorrhoeae strains. We studied gonorrhoea prevalence and antimicrobial resistance of N. gonorrhoeae among patients of STI clinics in 3 large cities in Indonesia. Methods In 2014, urogenital swabs were obtained from sexually active adults, mainly female sex workers, men who have sex with men, and male-to-female transgenders attending STI clinics in Jakarta, Yogyakarta, and Denpasar. In some patients additional rectal and pharyngeal swabs were taken. Diagnosis of N. gonorrhoeae infection was established by real time PCR targeting Opa genes. Culture isolates of N. gonorrhoeae were tested for antimicrobial susceptibility against doxycyclin, ciprofloxacin, azithromycin, ceftriaxone and cefixime by E-test™ using EUCAST breakpoints. Results We included 764 participants: 346 (45.3%) males, 321 (42.0%) females and 97 (12.7%) transgenders (median ages 27, 30, and 38 years). In total, 800 samples were collected: 443 urethral, 321 cervical, 23 rectal, and 13 pharyngeal swabs. In 215 (28.1%) participants N. gonorrhoeae infection was detected by PCR. Prevalence varied significantly by gender (26.0% in males, 32.1% in females, and 22.7% in transgenders, p = 0.04); and varied significantly by city: 14.5% in Denpasar, 31.5% in Jakarta and 27.5% in Yogyakarta (p = 0.015). Of 77 isolates tested for antimicrobial susceptibility, all were sensitive to azithromycin, ceftriaxone, and cefixime. In contrast, resistance to doxycyclin (98.7%) and ciprofloxacin (100%) was very common. Conclusion This is the first study since 2004 describing the epidemiology of gonorrhoea and antimicrobial resistant strains in Indonesia. Prevalence of gonorrhoea is very high among STI clinic visitors. In view of the extensive resistance against doxycyclin and ciprofloxacin, these antibiotics are not appropriate treatment options for gonorrhoea; instead, extended spectrum cephalosporins are advised. Disclosure of interest statement The study is funded by Indonesian government through Beasiswa Unggulan (The Excellence Scholarship Program), Ministry of Education and Culture Republic of Indonesia and Public Health Service (GGD) of Amsterdam, The Netherlands. The authors declare that there is no conflict of interest.

Macacine Herpesvirus 1 in Long-Tailed Macaques, Malaysia, 2009–2011

Macacine herpesvirus 1 (MaHV1; B virus) naturally infects macaques (Macaca spp.) and can cause fatal encephalitis in humans. In Peninsular Malaysia, wild macaques are abundant, and translocation is used to mitigate human-macaque conflict. Most adult macaques are infected with MaHV1, although the risk for transmission to persons who handle them during capture and translocation is unknown. We investigated MaHV1 shedding among 392 long-tailed macaques (M. fascicularis) after capture and translocation by the Department of Wildlife and National Parks in Peninsular Malaysia, during 2009-2011. For detection of MaHV1 DNA, PCR was performed on urogenital and oropharyngeal swab samples. Overall, 39% of macaques were shedding MaHV1 DNA; rates of DNA detection did not differ between sample types. This study demonstrates that MaHV1 was shed by a substantial proportion of macaques after capture and transport and suggests that persons handling macaques under these circumstances might be at risk for exposure to MaHV1.

The prevalence and clinical significance of Chlamydia infection in island and mainland populations of Victorian koalas (Phascolarctos cinereus).

Chlamydia infection is known to impact the health of koalas (Phascolarctos cinereus) in New South Wales (NSW) and Queensland, but the clinical significance of Chlamydia infections in Victorian koalas is not well described. We examined the prevalence of Chlamydia infection and assessed associated health parameters in two Victorian koala populations known to be Chlamydia positive. The same testing regimen was applied to a third Victorian population in which Chlamydia had not been detected. We examined 288 koalas and collected samples from the urogenital sinus and conjunctival sacs. Detection and differentiation of Chlamydia species utilized real-time PCR and high-resolution melting curve analysis. Chlamydia pecorum was detected in two populations (prevalences: 25% and 41%, respectively) but only from urogenital sinus swabs. Chlamydia was not detected in the third population. Chlamydia pneumoniae was not detected. Chlamydia pecorum infection was positively associated with wet bottom (indicating chronic urinary tract disease) in one Chlamydia-positive population and with abnormal urogenital ultrasound findings in the other Chlamydia-positive population. The prevalence of wet bottom was similar in all populations (including the Chlamydia-free population), suggesting there is another significant cause (or causes) of wet bottom in Victorian koalas. Ocular disease was not observed. This is the largest study of Chlamydia infection in Victorian koalas, and the results suggest the potential for epidemiologic differences related to Chlamydia infections between Victorian koalas and koalas in Queensland and NSW and also between geographically distinct Victorian populations. Further studies to investigate the genotypes of C. pecorum present in Victorian koalas and to identify additional causes of wet bottom in koalas are indicated.

English

Awareness of the threat of methicillin resistant Staphylococcus aureus (MRSA) is growing. Oxacillin and methicillin are penicillinase-stable penicillins, and strains that are oxacillin and methicillinn resistant are historically termed MRSA. To determine the existence of MRSA strains patients attending treatments in hospitals in Imo state, 200 clinical specimens were examined using conventional method such as culture and sensitivity. The specimens include urogenital swabs, nasal swabs, wound swabs, pus and blood. Out of the 200 samples collected, 23.68% showed resistance to oxacillin, 25.00% of the isolates from Owerri Zone were MRSA, while 23.81% of isolates from Okigwe Zone yielded MRSA, and 20.00% from Orlu Zone yielded MRSA. The mean for zones is 25.33 ± 13.05. Children under the age of 10 have the highest incidence of (57.14%) of S. aureus isolates resistant to oxacillin, followed by the elderly people of age group 51 to 60 years (27.27%). The young adults between the ages of 21 to 30 have the least incidence (10.00%) and the mean for age groups is 43.66 ± 6.26. The mean for clinical specimens is 15.20 ± 22.16. Wound specimens produced the highest incidence (40.00%) among clinical specimens followed by pus (33.33%) and blood has the least occurrence (00.00%). Males produced the highest incidence between the sexes (26.67%) and women (21.74%), and the mean for sexes is 38.00 ± 11.31. This shows that MRSA exists in hospitals in Imo state and considering the danger, it portends to healthcare setting, and efforts are needed to contain its spread. Key words: Methicillin, antibiotics, zones, sex, children, specimen, infections.

Isolation of Mycoplasma genitalium from patients with urogenital infections: first report from the Latin-American region.

Isolation of Mycoplasma genitalium from clinical specimens remains difficult. We describe a modified culture system based on Vero cells grown in medium 199 with 2% foetal bovine serum (FBS). The culture system was evaluated using early passage M. genitalium strains M6271 and M6311 with growth monitoring by quantitative TaqMan PCR. Eleven endocervical swabs and one male urethral swab positive by 16S rRNA and MgPa1–3 PCRs were quantified and inoculated into Vero cell suspensions in medium 199 supplemented with 2% FBS and antibiotics. Cultures were incubated for 14 days. Cell passages and growth monitoring by TaqMan PCR were performed until the growth of M. genitalium reached ≥106 geq/mL. Confirmation of the new M. genitalium strains was performed by sequencing a 281 bp fragment of mgpB. The growth of Mycoplasma genitalium strains was recorded for all urogenital swab specimens in the modified cell-culture system. Growth of M. genitalium was obtained within 2 months and yielded 12 M. genitalium strains with all 11 isolates from females of an identical, but unique genotype. To our knowledge, this is the first successful isolation of M. genitalium in the Latin-American region. The use of Vero cell culture in 199 medium with 2% FBS is a method comparable to the Ultroser G culture system for isolation of M. genitalium. Genotyping of clinical samples and isolates should be performed to document the absence of cross-contamination.

P2.047 Evaluation of a Multiplex Real-Time PCR Assay For Rapid Detection ofC.trachomatisandN. Gonorrhoeaefrom Genital Clinical Specimens

Background Sexually transmitted diseases (STDs) are among the most common causes of illness in the world, being associated to acute disease, infertility, long term disability and death. In particular the prevalence of C.trachomatis and N. gonorrhoeae infections appear to remain steady or increasing, although both diagnostic and therapeuthical tools are available, at least in developed countries. Standard diagnostic protocols can be complex, time-consuming, and sensitivity of culture methods affected by specimen characteristics. In addition, certified methods are today required, especially in the management of cases of sexual abuse. Methods Multiplex real-time PCR method (Xpert CT/NG, Cepheid) has been tested versus N. gonorrhoeae standard culture and C. trachomatis molecular assay (Artus C. trachomatis TM PCR Kit, Qiagen). 45 clinical samples (female and male urogenital swabs, ocular infections, first void of urine, seminal fluid, and external quality control samples - UK NEQAS) were selected and stored at - 20°C. Xpert CT/NG allowed to perform in one-step extraction, amplification and detection of C.trachomatis and N. gonorrhoeae DNA directly from all the variety of selected clinical specimens, within 60 min. Results 100% results agreement was found between Xpert CT/NG and standard protocols, including expected results by UK-NEQAS quality assessment specimens. However, batch PCR is a multi-step time-consuming process, the turn-around-time (TAT) required from 8 to 48 hrs versus the single technical and computer-assisted interpretation step by Xpert CT/NG, resulting in a Conclusions Xpert CT/NG test is a easy, rapid, accurate and certified method in routine diagnostics of N. gonorrhoeae and C. trachomatis infections.

Development of a high-throughput fluorescence assay for detecting SNPs in hemostasis and folate metabolism genes for clinical use

Genetic predisposition of an individual patient should be taken in account to choose the proper treatment. Implementation to clinical practice requires the development of rapid, high-throughput, and easy assays intended to detect single nucleotide polymorphisms. A detection kit intended to identify the hemostasis and folate cycle gene mutations G20210A FII, G1691A FV, G10976A FVII, G103T FXIII, C807T ITGA2, T1565C ITGB3, 5G(-675)4G PAI, G(-455)A FGB, C677T and A1298C MTHFR, A2756G MTR, A66G MTRR was suggested in this work. The method is based on the polymerase chain reaction and subsequent melt curve analysis of the complexes of amplicons with specific probe. Three single nucleotide polymorphisms can be identified in one tube using our detection kit that increases the productivity of the analysis in the clinical use. Different types of biological samples (buccal epithelium, saliva, plasma, serum, and urogenital swabs) can be used as the initial material for DNA isolation and further analysis by the method developed in this work.

The First Report of Otarine Herpesvirus-1-Associated Urogenital Carcinoma in a South American Fur Seal (Arctocephalus australis)

Otarine herpesvirus (OtHV)-1-associated urogenital carcinoma has been well documented in the California sea lion (Zalophus californianus, CSL), but this is the first report of this tumour in a captive South American fur seal (Arctocephalus australis, SAFS). The gross and microscopical morphology of the tumour in the SAFS was identical to that described previously in CSLs and the tumour in the present case had metastasized within the urogenital tract and draining lymph nodes and to the lungs and one kidney. Immunohistochemistry revealed intra- and extracytoplasmic labelling of herpesvirus antigen in the cells of the tumour tissue and transitional epithelium of the urethra. OtHV-1 nucleic acids were detected within tumour tissue and from a urogenital swab by polymerase chain reaction. The ranges of these two species of pinniped do not overlap normally in the wild, suggesting that transmission of OtHV-1 probably occurred in captivity. This confirmed susceptibility of the SAFS to the development of OtHV-1-associated urogenital carcinoma suggests that all species of Otariidae should be screened for OtHV-1 infection prior to movement within and between zoological collections.