Comparison of cervicovaginal fluid extracellular vesicles isolated from paired cervical brushes and vaginal swabs.

Endometriosis is a common gynaecological condition, with a long diagnostic delay. Surgery is required to confirm a diagnosis, highlighting the need for a non-invasive biomarker. Extracellular vesicles (EVs) may have a role in endometriosis pathogenesis, yet there is limited EV biomarker literature available. This study aimed to investigate the feasibility of isolating cervico-vaginal fluid EVs sampled using cervical brushes and vaginal swabs and to compare these methods. After providing informed consent, patients undergoing surgery for suspected endometriosis had cervical brush and vaginal swab samples collected under general anaesthetic. Isolated EVs were characterised through negative stain transmission electron microscopy (TEM), Western blotting (TSG101, CD63, Calnexin, ApoB, Albumin), tunable resistive pulse sensing (TRPS), microBCA assays and RT-qPCR of miRNAs. PCR was performed on samples prior to EV isolation to assess bacteria present in samples. Cervical brush and vaginal swab EVs were intact vesicles with limited co-isolated contaminants. Cervical brushes had higher concentrations of particles compared to match vaginal swabs, although both samples had low concentrations. Protein and miRNA yield were similar between matched samples. PCR demonstrated only a small amount DNA within samples was bacterial (>0.5%). Cervico-vaginal fluids EVs were successfully isolated from cervical brushes and vaginal swabs, demonstrating a new method of sampling reproductive EVs. EV yield from both sample types was low. Similar protein and miRNA levels suggest either sampling method may be suitable for biomarker studies.

Association of Mycoplasma canis with Fertility Disorders in Dogs: A Case Study Supported by Clinical Examination, PCR, 16S Microbiota Profiling, and Serology.

The role of Mycoplasma canis in canine fertility disorders is still poorly understood. As infection is often asymptomatic, there is an increasing need for appropriate diagnostic methods and treatment plans that would allow the reliable detection of M. canis infection and rapid alleviation of infection symptoms in affected dogs. In this study, we included 14 dogs with fertility problems and 16 dogs without fertility disorder signs. We compared clinical examination data and selected laboratory parameters (hematology and biochemistry) between the groups. We performed PCR-based detection of M. canis and 16S rRNA gene-based microbiota profiling of DNA isolated from vaginal and preputial swabs. Dog sera were tested for the presence of M. canis-specific antibodies. Hematological and selected biochemical parameters showed no differences between groups. PCR-based detection of M. canis in the samples was consistent with the results of 16S microbiota profiling. Several other bacterial taxa were also identified that could potentially be involved in different fertility disorders. Serological methods were not accurate enough since high cross-reactivity rates were observed. In the future, more accurate and efficient methods will be needed to determine the role of M. canis and its true role in the pathogenesis of specific fertility disorders in dogs.

Molecular epidemiology of Candida africana isolates collected from vagina swabs in French Guiana.

Previous molecular studies have shown that Candida africana corresponds to the clade 13 of Candia albicans. It has been mostly involved in vulvovaginal candidiasis worldwide but few data exist in South America. The aim of our study was to investigate the prevalence of C. africana in women living in French Guiana. For this, we first set up a fluorescent-intercalating-dye-real time Polymerase Chain Reaction (PCR) targeting the hyphal wall protein 1 gene. The test was applied to 212 C. albicans isolates collected from May to August 2019 from vaginal swabs, allowing the identification of six women harboring C. africana (eight isolates). The in vitro susceptibility of these eight isolates to six antifungal drugs was also evaluated. No demographics or clinical-specific features could be demonstrated. Genetic diversity of those isolates was analyzed through multilocus sequence typing and showed that diploid sequence type 182 was predominant (n = 6) and allowed the report of a new diploid sequence type.

Candida africana, the clade 13 of C. albicans, is characterized by specific genetic and phenotypic traits. Using a new molecular technique, we report a high prevalence of C. africana in vaginal swabs from patients living in French Guiana. The worldwide predominant genotype was detected in all but one patient.

Menstrual Tampons Are Reliable and Acceptable Tools to Self-Collect Vaginal Microbiome Samples.

Many women report embarrassment as the cause for their avoidance of routine gynaecological screening appointments. Methods of self-collection of bio samples would perhaps encourage women to participate in routine screening programs. The vaginal microbiome plays a key role in women's health and reproductive function. Microbial disturbances can result in the loss of lactobacillus dominance, also known as dysbiosis, associated with an increased risk of contracting sexually transmitted infections (STIs), pregnancy complications and infertility. Our primary aim was to determine if vaginal microbiome screening results are comparable between two methods for self-collected sample acquisition: tampons and lower vaginal swabs (LVSs). Secondary aims included the assessment of the effect of pre-analytic storage on the data (to streamline processing), the prevalence of dysbiosis and the acceptability of the tampons to the participants. Statistical analysis revealed no significant difference in the microbiome data, from tampons versus LVSs or fresh versus frozen samples. The prevalence of dysbiosis in this population of healthy volunteers was 42.9%. The questionnaire data revealed that 52.4% of volunteers use tampons every period, and the majority of volunteers rated the tampons as 5 on a 1-5 Likert scale regarding their perceived comfort using tampons. All (100%) of volunteers were happy to provide a tampon as a sample for testing. The findings from this study show that tampons and LVSs were comparable when analysing the vaginal microbiome, with potential superiority of the tampon with regard to patient acceptability. Self-collection of vaginal secretions for gynaecological screening using tampons warrants further research as this could change the screening landscape, ensuring wider participation and increasing efficacy.

Feasibility of using volatile urine fingerprints for the differentiation of sexually transmitted infections.

Sexually transmitted infections (STIs) are a public health problem worldwide, and current diagnostic methods have certain limitations. In recent years, volatile organic compounds (VOCs) have been studied as an alternative diagnostic method. Due to this, this study aimed to detect, in vaginal swabs and urine samples, VOCs emitted by highly prevalent STIs-causing bacteria (Chlamydia trachomatis, Mycoplasma genitalium, and Neisseria gonorrhoeae) to identify potential biomarkers that allow the detection of these STIs. VOCs detected in urine samples showed a better differentiation of patients with STIs due to C. trachomatis from those not infected, with 2,6-dimethyl-4-heptanone as the volatile compound most related to the presence of this bacterium. Among the VOCs most related to M. genitalium in urine, 4-methyltetradecane and 2-methylpentadecane stood out, while 3,4,4-trimethyl-2-cyclohexen-1-one was the VOC most closely related to N. gonorrhoeae infection. Moreover, C12 alcohols were the main VOC family associated with positive samples in all three bacteria, which could indicate the presence of aldehyde reductases in their metabolism. In contrast, alcohols such as 3-methyl-1-heptanol and 1-octanol, as well as dimethyl esters, were more associated with negative samples and may be useful in ruling out an STI caused by one of these three bacteria. In short, the VOCs identified as potential biomarkers in patients with infection by C. trachomatis, M. genitalium, or N. gonorrhoeae could be used in the early diagnosis of these STIs, quickly interrupting the chain of transmission, especially interesting in asymptomatic patients. KEY POINTS: • Sexually transmitted infections are a serious public health problem worldwide. • The study of VOCs in multiple infections is increasing in recent years. • The identification of volatile biomarkers could allow new diagnostic methods.

The Impact of the Microbiological Vaginal Swab on the Reproductive Outcome in Infertile Women.

BACKGROUND: The thesis on which this paper is based intended to investigate whether the result of the microbiological vaginal swab has an influence on the outcome of the fertility treatment. METHODS: The microbiological vaginal swabs of patients who received fertility treatment at Saarland University Hospital were evaluated. Depending on the microorganisms detected, the swab result was classified as inconspicuous, intermediate, or conspicuous. The SPSS software was used to determine the correlation between the swab result and the outcome of the fertility treatment. RESULTS: Dysbiosis was associated with a worse outcome of fertility treatment. The pregnancy rate with a conspicuous swab was 8.6%, whereas it was 13.4% with an inconspicuous swab. However, this association was not statistically significant. Furthermore, an association of endometriosis with dysbiosis was found. Endometriosis was more frequent with a conspicuous swab result than with an inconspicuous result (21.1% vs. 17.7%), yet the correlation was not statistically significant. However, the absence of lactobacilli was significantly associated with endometriosis (p = 0.021). The association between endometriosis and a lower pregnancy rate was also statistically significant (p = 0.006). CONCLUSION: The microbiological vaginal and cervical swabs can be used as predictors for the success of fertility treatments. Further studies are needed to assess the impact of transforming a dysbiotic flora into a eubiotic environment on the success of fertility treatments.

A pilot clinical validation study of a self-collected vaginal swab device for the detection of chlamydia trachomatis in women.

Chlamydia trachomatis (C. trachomatis) is one of the most prevalent preventable sexually transmitted diseases (STDs) in the world. In women, C. trachomatis infection can lead to long-term complications such as pelvic inflammatory disease (PID), and other related conditions such as ectopic pregnancies and even tubal factor infertility. These complications are preventable given early detection and clinical intervention, but these efforts are often hampered by asymptomatic silent infections, and non-compliance to screenings for STDs. Some women do not get tested out of concerns for violation of privacy, and fear of discomfort. Clinicians often use a multitude of tests to determine if a patient is infected by C. trachomatis, including a Polymerase Chain Reaction (PCR) test of First catch urine (FCU) samples. However, these tend to be inconvenient to store and transport, as they carry risk of spillage and have stringent refrigeration requirements. Moreover, given the gold-standard recommendations set forth by the Centres for Disease Control (CDC), the current technique can be inconvenient in remote areas where refrigeration and transport may not always be reliable. The current study therefore looks at the potential of a self-collected vaginal swab device that relies on Nucleic Acid Amplification Tests (NAATs), is dry-stored, and does not require refrigeration, to detect the presence of C. trachomatis in women. The study found evidence to suggest that the self-collection device has the potential to aid clinicians in the diagnosis of C. trachomatis in women when compared to doctor-collected vaginal discharge samples as the designated standard, FCU, and blood serology. Moreover, as a self-collection device it has the potential to break down some of the barriers to STD screening especially in young women, such as violation of privacy. The device therefore has a potential to encourage screening and therefore a potentially effective tool in the fight against the spread of preventable sexually transmitted diseases.

Prevalence and species distribution of microorganisms isolated among non-pregnant women affected by vulvovaginal candidiasis: A retrospective study over a 20 year-period.

BACKGROUND: Vulvovaginal candidiasis (VVC) is the second most common infection of the genital tract affecting millions of women worldwide. Data concerning the distribution and antifungal resistance of Candida species responsible of VVC vary among countries and population studied. OBJECTIVES: The aim of this work was to determine the prevalence, species distribution and antifungal susceptibility patterns of Candida species among symptomatic women over a 20-year period. METHODS: A total of 5,820 unique samples were retrospectively identified. Out of them, 1,046 (18%) were diagnosed with VVC. RESULTS: Women between 18 and 30 years had the highest prevalence rate of VVC (21%). Women aged less than 18 years and greater than 51 years had the highest prevalence rates of vaginal bacterial infections. Thirty-five (3.3%) women presented recurrent VVC. The most common yeast isolated was C. albicans, followed by C. glabrata, C. krusei, and C. parapsilosis. Non-Candida albicans species (NAC) were more significantly isolated among women aged 51 or above, than in women included in other groups (p < 0.01). Resistance to fluconazole and amphotericin B was infrequent in C. albicans strains. Resistance to fluconazole and amphotericin B was infrequent in C. albicans strains. NAC species presented higher resistance rates against fluconazole (30%) and voriconazole (25%). C. krusei and C. glabrata isolates showed lower MICs than most of the strains against amphotericin B (1 mg/L) and flucytosine (1 mg/L). CONCLUSIONS: Our findings indicated that continued surveillance on Candida species distribution and non-susceptibility rates to antifungals should be routinely reported to help the selection of the most appropriate drug, to avoid the emergence of resistant strains, and to improve the patient's outcomes.

Diagnostic accuracy of procalcitonin in maternal plasma to detect early intra-amniotic infection in preterm premature rupture of the membranes with respect of highvaginal swab as gold standard.

OBJECTIVE: To determine the diagnostic accuracy of procalcitonin in maternal plasma to detect early intra-amniotic infection in Preterm premature rupture of the membranes (PPROM) with respect of high vaginal swab as gold standard. METHODS: A cross-sectional study was conducted at Liaquat National Hospital, Karachi, from February to August 2017. The blood sample of women with PPROM were collected to measure procalcitonin level. PCT1 and PCT2 were run along with the sample for the accuracy of the results. Sensitivity, specificity, PPV, NPV, and diagnostic accuracy of procalcitonin were calculated taking HVS C/S as gold standard. RESULTS: Out of total 150 women, mean age was 28.78±4.79 years. Mean gestational age was 30.79±3.07 weeks. Mean procalcitonin level was 0.13±0.24 ng/ml. Intra-amniotic infection was diagnosed in 48.7% cases through procalcitonin levels and 51.3% through HVS culture and sensitivity. Sensitivity, Specificity, PPV (Positive predictive value), NPV (Negative predictive value) and accuracy were 87%, 91.8%, 91.78%, 87%, and 89.3% respectively. For females with gestational age ≤32 weeks, sensitivity, specificity, and diagnostic accuracy were 83.9%, 90.4%, and 87.03% respectively. For females with gestational age >32 weeks, sensitivity, specificity, and diagnostic accuracy were 95.2%, 92.5%, and 95.23% respectively. CONCLUSION: Diagnostic accuracy of maternal blood procalcitonin levels were found satisfactory in detection of early intra-amniotic infection in PPROM.

Detection of severe acute respiratory syndrome corona virus 2 in cervico-vaginal secretion of COVID-19-affected female: A prospective observational study from India.

BACKGROUND: The novel severe acute respiratory syndrome corona virus 2 (also known as 2019-nCoV) is a highly infectious agent and is declared as a global public health emergency by the World Health Organisation. The main known transmission route of severe acute respiratory syndrome corona virus 2 is through respiratory air droplets. Although recent studies have revealed that the virus is detectable in the throat, blood, urine, anal swabs, tears and even faeces; however, modes of transmission other than respiratory droplets has not been studied much. Knowledge on the presence of the virus in the female genital tract may help determine the risk of sexual transmission as well as the risk of mother-to-child transmission. However, not much data are available yet regarding the presence of the virus in the female genital system. Hence, to explore the presence of the virus in the female genital system and possibility of sexual transmission, a study was conducted where in we tried to detect severe acute respiratory syndrome corona virus 2 in cervico-vaginal secretions. METHODS: From July 2020 to September 2020, 35 COVID-19-positive female patients admitted to tertiary care teaching institute of Eastern India, which is now declared dedicated Corona Hospital and Centre of Excellence for COVID-19 care, who consented for the research were enrolled in this prospective observational study. Proper gynaecological history, clinical records along with laboratory findings of the patient was recorded. The possibility of the sexual transmission of the virus from female to her male partner was to be ascertained by testing the presence of severe acute respiratory syndrome corona virus 2 in the vaginal, cervical secretions by reverse transcriptase polymerase chain reaction. RESULTS: All 35 COVID-19-positive female patients were tested for severe acute respiratory syndrome corona virus 2 in their vaginal and cervical secretions by reverse transcriptase polymerase chain reaction. All the samples were tested negative for the virus. CONCLUSION: Findings from this study reveals that severe acute respiratory syndrome corona virus 2 is not present in the cervical and vaginal secretions, and the possibility of transmission from female to her male partner by vaginal sexual intercourse is unlikely.

Contribution of vaginal culture to predict early onset neonatal infection in preterm prelabor rupture of membranes.

BACKGROUND: Preterm prelabor rupture of membranes (PPROM) is a major cause of morbidity and mortality for both the mother and the newborn. The vaginal germ profile in PPROM is poorly known, particularly regarding the risk of early-onset neonatal infection (EONI). OBJECTIVE: To determine microbiological risk factors for EONI in case of PPROM before 34 weeks of gestation (WG). STUDY DESIGN: A retrospective single-center cohort of patients with PPROM before 34 W G from 2008 to 2016. Vaginal swabs were obtained at admission and at delivery as per usual care and were analyzed by Gram stain and culture for vaginal dysbiosisi.e lactobacilli depletion and/or presence of potential pathogens. RESULTS: Among 268 cases of PPROM, 39 neonates had EONI 14.55 %; (95 %CI 0.11 - 0.19) Overall, vaginal samples culture was positive in 16.67 % (95 %CI 11.95 %-22.32 %) at the time of rupture and 24.76 % (95 %CI 19.02 %-31.23 %) at delivery, with no significant differences between EONI and no-EONI groups (p = 0.797 and 0.486, respectively), including for Group B Streptococci (GBS) and Escherichia coli. EONI was significantly associated with dysbiosis at the time of rupture (23.94 % versus 10.35 % in the absence of dysbiosis, p = 0.009) and at delivery (19.70 % versus 3.90 % if no dysbiosis, p < 0.001). Clinical intra-uterine infection was present in 78.5 % (n = 31) of the EONI group versus 37.2 % (n = 85) in the non-EONI group (p < 0.001) and chorioamnionitis and/or funisitis were found in 97.3 % and 91.9 %, respectively in the EONI group, versus 56.11 % and 53.96 %, respectively, in the non-EONI group (p < 0.001). CONCLUSION: Dysbiosis following rupture and at delivery, but not the presence of pathogens in the VS culture, was associated with the risk of EONI in case of PPROM.

Self-obtained vaginal swabs are not inferior to provider-performed endocervical sampling for emergency department diagnosis of Neisseria gonorrhoeae and Chlamydia trachomatis.

OBJECTIVE: Provider-performed endocervical sampling (PPES) in the diagnosis of Neisseria gonorrhoeae (NG) and Chlamydia trachomatis (CT) may be difficult to perform in a busy emergency department (ED) due to patient preference, availability of the pelvic examination room, or provider availability. Our objective was to assess if self-obtained vaginal swabs (SOVS) were noninferior to PPES in the ED diagnosis of NG/CT using a rapid nucleic acid amplification test (NAAT). METHODS: We conducted a prospective observational cohort study in a single ED. Participants were adult female English- and Spanish-speaking patients in whom the ED provider felt that NG/CT testing was warranted. Each patient had SOVS and PPES performed. For SOVS, a research associate reviewed a one-page handout describing the procedure but gave no other assistance. Patients answered survey questions regarding acceptability of SOVS and symptomatology. We established a minimum sensitivity of 90% for SOVS to be considered clinically noninferior to standard PPES. RESULTS: A total of 533 patients completed enrollment and answered survey questions, 515 of whom had laboratory results for both SOVS and PPES. There were 86 patients with a positive result: 29 with NG, 47 with CT, and 10 with coinfection. SOVS had a sensitivity of 95% (95% confidence interval = 88% to 99%) for the detection of NG/CT when compared to PPES. SOVS were felt to be an acceptable collection method in 93% of patients and 75% preferred SOVS to PPES. CONCLUSION: SOVS are noninferior to PPES in NG/CT diagnosis using a rapid NAAT in ED patients and surveys indicate high patient acceptability.

Positive Reinforcement Conditioning as a Tool for Frequent Minimally Invasive Blood and Vaginal Swab Sampling in African Lions (Panthera Leo).

Information regarding the physiology of African lions is scarce, mainly due to challenges associated with essential routine research procedures. The aim of this experiment was to test the possibility of training six captive lionesses by positive reinforcement conditioning (PRC) to voluntarily allow the collection of vaginal swabs and blood samples. This was done with the final goal of avoiding frequent anesthesia, and potentially stressful management during reproduction research. All lionesses mastered basic clicker and targeting principles within 2 weeks. Routine sampling was possible after 20 weeks of training, enabling the collection of about 750 vaginal swabs and 650 blood samples over 18 months. The animals remained calm and cooperative during all sessions, and demonstrated curiosity in the training. PRC training of captive lionesses proved to be a suitable, minimally invasive method for repeated collection of vaginal swabs and blood. Additionally, PRC may serve as behavioral enrichment for African lions in captive settings. Compared to chemical or physical restraining methods, this noninvasive management approach may reduce distress and physiological negative side effects, thus opening up new avenues for feline research.

Loop-Mediated Isothermal Amplification (LAMP) for Detection of Ureaplasma diversum from Cervico-vaginal Swab of Buffaloes.

The main plan of the current study was to develop a rapid, robust, and field-applicable loop-mediated isothermal amplification (LAMP) assay for the detection of Ureaplasma diversum. A strain-specific 16S rRNA gene of Ureaplasma diversum was used for detection which was cloned, sequenced, and characterized earlier. LAMP results were visualized within 90 min with the naked eye. Cervico-vaginal swabs of 50 buffaloes were randomly collected from Livestock Research Center of NDRI as per the Institute Animal ethics guidelines. Out of 50 cervico-vaginal swab samples collected randomly, 34 were found positive with LAMP while 16 samples were negative. Conventional PCR results showed the same result. Therefore, the accuracy of the developed LAMP was about 100%. The developed LAMP assay can also be used to screen the animals for Ureaplasma diversum infection in cervico-vaginal swab. However, further study is needed to assess sensitivity and accuracy towards their detection and their relationship in disease diagnosis.

Diagnostic performance of direct latex agglutination, post-enrichment latex agglutination and culture methods in screening of group B streptococci in late pregnancy: a comparative study.

BACKGROUND: Group B streptococcus (GBS) is one of the main causes of neonatal sepsis. PURPOSE: Evaluation of the diagnostic performance of direct latex agglutination test (DLA), post-enrichment latex agglutination (LA) test, and direct culture on chromogenic media in rapid identification of GBS carrier in pregnant women in comparison with the conventional post-enrichment CDC-recommended culture method and further to estimate GBS carriage prevalence and its antimicrobial susceptibility. METHODS: Two hundred pregnant women at gestational age (35-37 weeks) were enrolled. Three low vaginal swabs were obtained from each participant. One swab was directly inoculated into Strep B Select (SBS) agar. The second swab was inoculated in enrichment Lim broth for immunological antigen detection by post-enrichment latex agglutination (5 h and 24 h) and subculture for bacteriological detection. The third swab was used for immunological detection of GBS antigen by direct latex agglutination. The isolated GBS was subjected to antimicrobial susceptibility testing. RESULTS: Among 200 pregnant women, 47 (23.5%) were GBS carriers. Considering post-enrichment subculture on SBS medium as a gold standard, the sensitivities for post-enrichment 5 h and 24 h LA were 66% and 95.7%, respectively. However, direct cultivation of the vaginal swabs on SBS medium and DLA recorded 83% and 4.3%, respectively, for sensitivity. All GBS isolates (100%) were sensitive to penicillin G, ampicillin, ceftriaxone, and vancomycin. In contrast, 21.3% and 12.8% of isolated GBS were resistant to erythromycin and clindamycin, respectively. CONCLUSION: Group B streptococcal antigen detection by latex agglutination after 5 h enrichment is a reliable, easy, and relatively rapid method for screening of GBS carriage in pregnant woman not in labor. Latex agglutination after 18-24 h enrichment can be used alternative to standard subculture method for screening GBS carriage.

Pregnancy-related listeriosis: frequency and genotypic characteristics of L. monocytogenes from human specimens in Kerman, Iran.

Listeria monocytogenes is a foodborne pathogen that can pose serious complications during pregnancy and neonatal infection. This study aimed to determine the frequency of L. monocytogenes infection, prevalent serotypes, and virulence genes among pregnant women and those experiencing miscarriages in Kerman, Iran. Out of 200 vaginal swabs, 4.5 and 29.5% of specimens were positive for L. monocytogenes infection as identified by culture and molecular methods, respectively. The majority of isolates from positive cultures (89%) of pregnant women resulted in stillbirth, death, and blindness. The most prevalent virulence determinants were inl B, prf A, and act A. The majority of isolates were non-typable. A history of miscarriage and gestational age are known to be significantly associated with the presence of infection. This study emphasizes the importance of initial screening for L. monocytogenes in pregnant women in Iran. Molecular methods may be useful in this process. Increasing the awareness of pregnant women could be effective in reducing pregnancy-related listeriosis.

A paperfluidic platform to detect Neisseria gonorrhoeae in clinical samples.

Globally, the microbe Neisseria gonorrhoeae (NG) causes 106 million newly documented sexually transmitted infections each year. Once appropriately diagnosed, NG infections can be readily treated with antibiotics, but high-risk patients often do not return to the clinic for treatment if results are not provided at the point of care. A rapid, sensitive molecular diagnostic would help increase NG treatment and reduce the prevalence of this sexually transmitted disease. Here, we report on the design and development of a rapid, highly sensitive, paperfluidic device for point-of-care diagnosis of NG. The device integrates patient swab sample lysis, nucleic acid extraction, thermophilic helicase-dependent amplification (tHDA), an internal amplification control (NGIC), and visual lateral flow detection within an 80 min run time. Limits of NG detection for the NG/NGIC multiplex tHDA assay were determined within the device, and clinical performance was validated retroactively against qPCR-quantified patient samples in a proof-of-concept study. This paperfluidic diagnostic has a clinically relevant limit of detection of 500 NG cells per device with analytical sensitivity down to 10 NG cells per device. In triplicate testing of 40 total urethral and vaginal swab samples, the device had 95% overall sensitivity and 100% specificity, approaching current laboratory-based molecular NG diagnostics. This diagnostic platform could increase access to accurate NG diagnoses to those most in need.

A nested-PCR strategy for molecular diagnosis of mollicutes in uncultured biological samples from cows with vulvovaginitis.

Bacteria classified in Mycoplasma (M. bovis and M. bovigenitalium) and Ureaplasma (U. diversum) genera are associated with granular vulvovaginitis that affect heifers and cows at reproductive age. The traditional means for detection and speciation of mollicutes from clinical samples have been culture and serology. However, challenges experienced with these laboratory methods have hampered assessment of their impact in pathogenesis and epidemiology in cattle worldwide. The aim of this study was to develop a PCR strategy to detect and primarily discriminate between the main species of mollicutes associated with reproductive disorders of cattle in uncultured clinical samples. In order to amplify the 16S-23S rRNA internal transcribed spacer region of the genome, a consensual and species-specific nested-PCR assay was developed to identify and discriminate between main species of mollicutes. In addition, 31 vaginal swab samples from dairy and beef affected cows were investigated. This nested-PCR strategy was successfully employed in the diagnosis of single and mixed mollicute infections of diseased cows from cattle herds from Brazil. The developed system enabled the rapid and unambiguous identification of the main mollicute species known to be associated with this cattle reproductive disorder through differential amplification of partial fragments of the ITS region of mollicute genomes. The development of rapid and sensitive tools for mollicute detection and discrimination without the need for previous cultures or sequencing of PCR products is a high priority for accurate diagnosis in animal health. Therefore, the PCR strategy described herein may be helpful for diagnosis of this class of bacteria in genital swabs submitted to veterinary diagnostic laboratories, not demanding expertise in mycoplasma culture and identification.

Comparative Study on the Vaginal Flora and Incidence of Asymptomatic Vaginosis among Healthy Women and in Women with Infertility Problems of Reproductive Age.

INTRODUCTION: The normal vaginal flora is highly complex, dominated by lactobacilli of doderlein that plays a vital role in maintaining the women's health and inhibits other pathogenic microorganisms. Fluctuation in local environment or exposure to any exogenous and endogenous sources changes the vaginal flora over a period of time. Disruption of the vaginal ecosystem changes the microflora of the healthy vagina, altering the pH and predisposing to lower reproductive tract infections. The change in the microflora of the female genital tract by pathogenic organisms may ascend from vagina to upper genital tract and may cause infertility. Although several studies demonstrate a higher prevalence of bacterial vaginosis in infertile population. The role of vaginal microbiome in infertility is not clear and need to be explored further. AIM: To compare the vaginal flora and analyse the incidence of asymptomatic vaginosis among healthy women and in women with infertility problems. MATERIALS AND METHODS: A cross-sectional study was conducted over a period of six months at Sri Lakshmi Narayana Medical College and Hospital Puducherry, India. A total of 200 high vaginal swabs were collected from Group 1 which included 84 healthy women with regular menstrual cycles without any gynaecological disorder and from Group 2, 116 women with infertility problems attending fertility clinic within the age group of 18 to 45 years. All swabs were subjected to routine aerobic, anaerobic and fungal culture. Saline wet mount was performed for the detection of clue cells and Trichomonas vaginalis, 10% KOH was performed for demonstration of budding yeast cells and pseudo hyphae, Gram's staining to determine the presence of yeast cells, leucocytes and bacterial morphotypes. The smear was also graded using Nugent scoring system. RESULTS: The vaginal flora of Group 1 was dominated by Lactobacillus (40, 27.8 %) followed by Micrococcus (22, 15.3 %), Enterococcus (16, 11.1%), Coagulase negative Staphylococcus spp. (12, 8.3%). Whereas in Group 2, the most dominant flora was Candida spp. (30, 26.5 %), Enterococcus (26, 23%) followed by Gram negative bacilli such as E. coli (16, 14.1 %). The percentage of Lactobacillus in Group 2 women with infertility problems was relatively low (4, 3.5%). Asymptomatic vaginosis was present in 32 (27.6 %) of Group 2 women compared to Group 1 women were only 6 (7.1%) had asymptomatic vaginosis. CONCLUSION: Women with infertility problems showed higher prevalence of asymptomatic vaginosis and abundance of Bacterial Vaginosis (BV) associated bacteria compared to healthy women. Hence, this study recommends the screening of vaginal flora as a routine for all women, especially in women undergoing infertility treatment and also suggests the importance of vaginal culture and sensitivity in routine practice.

High prevalence of human papillomaviruses in Ghanaian pregnant women.

Data about the prevalence of human papillomaviruses (HPV) in African women with normal and abnormal cervical cytology are still scarce. Current HPV vaccines contain HPV types, which mainly represent the HPV epidemiology of industrial countries. As further developments of HPV vaccines are going on, it is necessary to regard regional differences in HPV type prevalence to ensure optimal protection by the vaccine. Vaginal swabs of Ghanaian pregnant women, routinely collected before delivery to rule out bacterial infections causing early onset sepsis, were screened for 12 high-risk (HR), 13 probably/possibly (pHR), and 18 low-risk (LR) HPV types. Most pregnant women come for delivery to the hospital. This was considered as appropriate possibility to have an unselected group of women. HPV DNA were detected in 55/165 women (33.3, 95 % CI 26.3-41.1 %). Thirty-four out of fifty-five (61.8, 95 % CI 47.7-74.3 %) of HPV-positive women were infected with HR and/or pHR HPV types. The five most prevalent HR or pHR HPV types were HPV-52 and HPV-67 (7 women each, 4.2, 95 % CI 1.9-8.9 %), HPV-53 (six women, 3.6, 95 % CI 1.5-8.1 %), HPV-45 (five women, 3.0, 95 % CI 1.1-7.3 %), and HPV-18 (four women, 2.4, 95 % CI 0.8-6.5 %), respectively. HPV-16 was found in two women only (1.2, 95 % CI 0.2-4.8 %). Future HPV vaccine research may devote special interest to HPV-67 and HPV-53 provided further studies confirm their high prevalence in the general population of Sub-Saharan African countries. The true carcinogenic potential of HPV-67, which is a member of species alpha9 including HPV-16, and so far categorized as pHR, should be clarified.