Vaginal Secretion Samples Research Articles

Evaluations and comparisons of microbial diversities in four types of body fluids based on two 16S rRNA gene sequencing methods

BackgroundBody fluids are one of the common biological traces at crime scenes. Understanding the types of these biological traces could provide key clues for the investigations of the forensic cases. In recent years, partial hypervariable regions of 16S rRNA gene sequencing and full-length 16S rRNA gene sequencing have attracted the interests of researchers and we intend to explore which method can be better applied to forensic researches. MethodsIn this study, the 16S rRNA gene V3-V4 (short-read) sequencing based on next-generation sequencing and the full-length 16S rRNA gene sequencing based on single molecule real-time sequencing were used to classify microbes in saliva, peripheral blood, vaginal secretion and menstrual blood samples. ResultsAlpha diversity metrics in short-read sequencing were larger than those of full-length sequencing. Phylum-level bacteria in four kinds of body fluids obtained from the two platforms were similar, while their abundances were different. The results of principal coordinates analysis and analysis of molecular variance indicated the microbial compositions of vaginal secretion and menstrual blood samples were similar, and the microbial compositions among saliva, peripheral blood, vaginal secretion or menstrual blood samples were significantly different. The linear discriminant analysis effect size showed the differential bacteria screened among the four kinds of body fluids were variant in two sequencing results. ConclusionBoth sequencing methods could be used to detect bacterial diversities in four different types of body fluids and provide potential tools for microbes to identify the four kinds of body fluids in forensic investigation, in which full-length sequencing could provide more accurate taxonomy.

P–795 Assessment of the risk of contamination of semen, follicular and vaginal fluids with SARS-CoV–2 virus in patients undergoing ART

Abstract Study question Is SARS-CoV–2 detected by RT-PCR in the reproductive materials and follicular fluid of asymptomatic patients undergoing fertility treatments? Summary answer No SARS-CoV–2 mRNA was detected in sperm, vaginal and follicular fluids samples of asymptomatic patients, irrespective of the nasopharyngeal swab or COVID–19 questionnaire results. What is known already: The COVID–19 pandemic had a huge impact on health care including on fertility clinics. While activities were interrupted during the first wave, ART cycles are currently performed but uncertainties remain regarding the presence of the virus in reproductive materials. The SARS-CoV–2 receptors were detected in reproductive organs but only few studies with limited number of cases reported the presence of SARS-CoV–2mRNA in semen of symptomatic patients. In women, the risk of SARS-CoV–2 contamination in follicular and vaginal fluids remains uncertain. Thus the risk of sexual transmission and the safety of the IVF laboratory procedures are unclear. Study design, size, duration This COVART study is an observational cohort prospective trial conducted at a Belgian academic hospital. Between September 2020 and January 2021, 208 asymptomatic adults patients (men and women) undergoing ART treatments (sperm analysis, IUI, ICSI/ICF cycles, oocyte cryopreservation) were included in the trial after informed consent. All patients followed standard procedures to evaluate COVID–19 risk (nasopharyngeal swab during ovarian stimulation and COVID–19 risk questionnaire). Participants were divided into two groups: COVID–19 positive and negative/unknown groups. Participants/materials, setting, methods: Swabs on the residual reproductive materials were done and stored in viral transport medium at 4 °C until processing. After addition of an internal control in each sample and virus inactivation with Trizol, mRNA was extracted using phenol-chloroform method. Quantitative RT-PCR was performed in duplicate following a previously validated protocol (45 cycles, Roche Light Cycler 480). Negative/positive controls were used to validate each run. The test was considered as positive when CT < 40. Main results and the role of chance A total of 399 samples (126 semen, 162 vaginal fluid, 111 follicular fluid samples) of reproductive residual materials from 208 participants were collected during the peak of the second wave of COVID–19 pandemic, when Belgium was considered as a red zone with a viral Rt of 1.516 and a 14-day COVID–19 cases notification rate above 630 per 100000. Although the policy of the fertility clinic was to cancel all cycles of patients with a positive nasopharyngeal swab test except if specific medical raisons to continue the cycle, 14 samples from 9 non-cancelled patients diagnosed with COVID–19 before or just after the samples collection were analyzed (4 sperm, 5 follicular fluid and 5 vaginal secretion samples). For the 199 remaining patients, the COVID–19 status was negative or unknown. None of the samples were considered as positive after quantitative RT-PCR analysis. Limitations, reasons for caution All the patients were asymptomatic at the time of the samples collection and the large majority was negatively diagnosed for COVID–19 during the ART cycle. The results should be confirmed by including a larger cohort of positive patients. Data on the impact on ART outcomes will be evaluated. Wider implications of the findings: We showed that contaminations of sperm, follicular and vaginal fluids with SARS-CoV–2 are unlikely in asymptomatic patients, even when diagnosed positive, confirming the poor risk of sexual transmission. Moreover, no additional safety measures seems to be implemented in the IVF laboratory to ensure the safety of the staff. Trial registration number P2020/414

Two-Target Quantitative PCR To Predict Library Composition for Shallow Shotgun Sequencing.

ABSTRACTWhen determining human microbiota composition, shotgun sequencing is a powerful tool that can generate high-resolution taxonomic and functional information at once. However, the technique is limited by missing information about host-to-microbe ratios observed in different body compartments. This limitation makes it difficult to plan shotgun sequencing assays, especially in the context of high sample multiplexing and limited sequencing output and is of particular importance for studies employing the recently described shallow shotgun sequencing technique. In this study, we evaluated the use of a quantitative PCR (qPCR)-based assay to predict host-to-microbe ratio prior to sequencing. Combining a two-target assay involving the bacterial 16S rRNA gene and the human beta-actin gene, we derived a model to predict human-to-microbe ratios from two sample types, including stool samples and oropharyngeal swabs. We then validated it on two independently collected sample types, including rectal swabs and vaginal secretion samples. This assay enabled accurate prediction in the validation set in a range of sample compositions between 4% and 98% nonhuman reads and observed proportions varied between −18.8% and +19.2% from the expected values. We hope that this easy-to-use assay will help researchers to plan their shotgun sequencing experiments in a more efficient way.IMPORTANCE When determining human microbiota composition, shotgun sequencing is a powerful tool that can generate large amounts of data. However, in sample compositions with low or variable microbial density, shallowing sequencing can negatively affect microbial community metrics. Here, we show that variable sequencing depth decreases measured alpha diversity at differing rates based on community composition. We then derived a model that can determine sample composition prior to sequencing using quantitative PCR (qPCR) data and validated the model using a separate sample set. We have included a tool that uses this model to be available for researchers to use when gauging shallow sequencing viability of samples.

Vertical transmission of SARS-CoV-2: A prospective cross-sectional study from a tertiary center.

The aim was to investigate the association of the delivery mode and vertical transmission of severe acute respiratory syndrome coronavirus‐2 (SARS‐CoV‐2) through the samples of vaginal secretions, placenta, cord blood, or amniotic fluid as well as the neonatal outcomes. This cross‐sectional study presents an analysis of prospectively gathered data collected at a single tertiary hospital. Sixty‐three pregnant women with confirmed coronavirus disease 2019 (COVID‐19) participated in the study. Vertical transmission of SARS‐CoV‐2 was analyzed with reverse transcriptase‐polymerase chain reaction (RT‐PCR) tests and blood tests for immunoglobulin G (IgG)–immunoglobulin M (IgM) antibodies. All patients were in the mild or moderate category for COVID‐19. Only one placental sample and two of the vaginal secretion samples were positive for SARS‐CoV‐2. Except for one, all positive samples were obtained from patients who gave birth by cesarean. All cord blood and amniotic fluid samples were negative for SARS‐CoV‐2. Two newborns were screened positive for COVID‐19 IgG–IgM within 24 h after delivery, but the RT‐PCR tests were negative. A positive RT‐PCR result was detected in a neof a mother whose placenta, cord blood, amniotic fluid, and vaginal secretions samples were negative. He died due to pulmonary hemorrhage on the 11th day of life. In conclusion, we demonstrated that SARS‐CoV‐2 can be detectable in the placenta or vaginal secretions of pregnant women. Detection of the virus in the placenta or vaginal secretions may not be associated with neonatal infection. Vaginal delivery may not increase the incidence of neonatal infection, and cesarean may not prevent vertical transmission. The decision regarding the mode of delivery should be based on obstetric indications and COVID‐19 severity.

Vaginal Bacterial Profile in Pregnant Women

Summary This study aimed to determine the bacterial profile of vaginal samples from pregnant women and the prevalence of identified microorganisms. Materials and methods: A prospective epidemiological study was conducted on 150 pregnant women admitted to the Clinic of Obstetrics and Gynecology at Dr. G. Stranski University Hospital in Pleven. Vaginal secretion samples were collected from all the 150 women. Direct microscopy of native smears prepared by the wet mount method to detect Trichomonas vaginalis and direct microscopy of Gram-stained smears was performed. Aerobic culture studies were conducted to identify pathogenic bacteria and Sabouraud glucose agar (SGA) to detect the presence of Candida spp. Samples were also collected from 107 of the women for PCR tests for mycoplasma detection. Results: The highest relative share in the study was women aged 21 to 35 years 104 (69.33%). Of all the hospitalized and examined pregnant women, 87 (57.99%) had medical problems during the current pregnancy and postpartum complications, and 63 women (42%) gave birth without accompanying complications. In 86 (57.32%) of the women, bacteria were detected by direct microscopy of Gram-stained smears. Of these, 24 (16%) had gram-negative rods, 28 (18.66%) had gram-positive cocci. In 34 (22.66%), we found gram-negative and gram-variable pleomorphic bacteria. The aerobic culture study revealed microbiological growth in 44 (29.33%) of the women investigated. Conclusions: Studies on the profile of the vaginal microbiota in pregnant women and the spread of pathogenic microorganisms are essential for make prognoses for pregnancy outcome, evaluating the risk of developing a maternal-fetal infection, and discussing options for timely treatment.

Degradation of human mRNA transcripts over time as an indicator of the time since deposition (TsD) in biological crime scene traces

Knowledge about the age of a stain, also termed as time since deposition (TsD), would provide law-enforcing authorities with valuable information for the prosecution of criminal offenses. Yet, there is no reliable method for the inference / assessment of TsD available. The aim of this study was to gain further insight into the RNA degradation pattern of forensically relevant body fluids and to find candidate markers for TsD estimation. Blood, menstrual blood, saliva, semen and vaginal secretion samples were exposed to indoor (dark, room temperature) and outdoor (exposed to sun, wind, etc. but protected from rain) conditions for up to 1.5 years. Based on expression and degradation analyses, we were able to identify body fluid specific signatures and RNA degradation patterns. The indoor samples showed a marked drop in RNA integrity after 6 months, while the outdoor samples were difficult to interpret and therefore excluded for some of the analyses. Up to 4 weeks, indoor samples showed more stable and less degrading transcripts than outdoor samples. Stable transcripts tended to be significantly shorter than degrading ones or transcripts, which are neither degrading nor stable. We reinforced the body fluid specific and the housekeeping gene nature of previously reported markers. With an unbiased approach, we selected stable and degrading genes for each body fluid in the short term and assessed their integrity during extended storage. We identified several stable and degrading gene transcripts, which could be tested in a targeted assay to assess the TsD interval e.g. by analyzing the ratio of degrading vs stable transcripts. In conclusion, we were able to detect RNA degradation patterns in samples being aged up to 1.5 years and identified several candidate markers, which could be evaluated for TsD estimation.

DISTRIBUTION OF MYCOPLASMA SPP. AND UREAPLASMA SPP. AMONG PREGNANT WOMEN

The purpose of the present study is to determine the prevalence of genital mycoplasmas (M. genitalium, M. hominis, U. parvum, U. urealyticum) in pregnant women by molecular biological methods. Material/Methods: A prospective epidemiological study of 107 pregnant women hospitalized in the Clinic of Obstetrics and Gynecology, University Hospital-Pleven, Bulgaria, was conducted. Vaginal secretion samples were taken from all 107 pregnant women. A Polymerase chain reaction (PCR) assay was used to detect the genomic DNA of the bacteria in pregnant women. Results: The highest is the relative share of women in the age group from 20 to 35 years - 66 (64.68%), followed by women under 20 years - 27 (25.23%) and women over 35 years - 14 (13.08%). Detection of bacterial DNA was found in 85 (79.44%) of the cases, with present Ureaplasma spp. Colonization in 42 women (39.25%). Although no statistical dependence was found on open bacteria and age groups (p-value = 0.4688), it is noteworthy that the prevalence of Mycoplasma spp. and Ureaplasma spp. as a whole in the age group from 20 to 35 years, which has the highest birth rate, is more than twice higher than the group of up to 20 years and more than five times higher compared to the group over 35 years. Conclusions: Studies on the incidence of Mycoplasma spp. and Ureaplasma spp. in pregnant women is important for controlling the pregnancy, predicting the risk of developing maternal-fetal infection and discussing the options for timely treatment.

Comparison of viromes in vaginal secretion from pregnant women with and without vaginitis

BackgroundAlthough some studies have investigated the bacterial community in vaginal tract of pregnant women, there are few reports about the viral community (virome) in this type of microenvironment.MethodsTo investigate the composition of virome in vaginal secretion samples, 40 vaginal secretion samples from pregnant women with vaginitis and 20 vaginal secretion samples from pregnant women without vaginitis, pooled into 4 and 2 sample pools, respectively, were subjected to viral metagenomic analysis.ResultsResults indicated virus sequences showing similarity to human papillomavirus (HPV), anellovirus, and norovirus were recovered from this cohort of pregnant women. Further analysis indicated that 15 different defined types and one unclassified type of HPV were detected from pregnant women with vaginitis while only 3 defined types of HPV were detected in pregnant women without vaginitis. Five different groups of viruses from the family Anelloviridae were present in pregnant women with but none of them were detected in pregnant women without vaginitis. Norovirus was detected in 3 out of the 4 sample pools from pregnant women with vaginitis but none in the pregnant women without vaginitis. Twelve complete genomes belonging to 10 different types of HPV, and 5 novel anllovirus genomes belonging 2 different genera in Anelloviridae were acquired from these libraries, based on which phylogenetical analysis and pairwise sequence comparison were performed. Phageome in these samples was also briefly characterized and compared between two groups.ConclusionOur data suggested that virome might play an important role in the progression of vaginitis in pregnant women.

Effect of oral Lactobacillus rhamnosus GR-1 and Lactobacillus reuteri RC-14 on vaginal Group B Streptococcus colonization and vaginal microbiome in late pregnancy

To explore the effects of intervention with oral probiotic Lactobacillus rhamnosus GR-1 and Lactobacillus reuteri RC-14 on vaginal Group B Streptococcus (GBS) colonization, pregnancy outcome and vaginal microbiome in GBS-positive women in the third trimester of pregnancy. This study were conducted among 155 women in the third trimester of pregnancy with positive results of GBS culture in the Outpatient Department of Zhujiang Hospital from March to November, 2019. After excluding 32 patients who received lactobacillus intervention for less than 2 weeks or underwent postpartum GBS retesting, the women were divided into oral probiotics intervention group (60 cases) and non-intervention group (63 cases). According to the results of GBS retesting, the 60 women in the intervention group were divided into GBS-negative group (18 cases) and persistent GBS-positive group (42 cases). At the end of the intervention, the rates of negative GBS culture result were calculated and the pregnancy outcomes were compared. From 5 women randomly selected from the intervention group, samples of vaginal secretions were collected before and after the intervention for amplicon sequencing and bioinformatics analysis. At the end of the intervention, the GBS-negative rate in the intervention group was 30% (18/60), as compared with 23% (3/13) in the non-intervention group. Probiotic intervention significantly reduced the incidence of premature rupture of membranes (P < 0.05) and reduced the use of antibiotics during pregnancy (P < 0.05). OTU analysis of the vaginal secretions suggested probiotic intervention decreased the total sequence number and GBS sequence number, increased the species composition, and significantly decreased GBS abundance (P < 0.05). Probiotics intervention also significantly decreased the species abundance of Enterococcus, Staphylococcus and Streptococcus in the vaginal flora (P < 0.05). Intervention with oral probiotics can reduce vaginal GBS colonization in late pregnancy and improve the pregnancy outcome. Lactobacillus is capable of reducing the abundance of GBS and other pathogenic bacteria to improve the microbiome of vaginal flora.

Prebiotic Maltose Gel Can Promote the Vaginal Microbiota From BV-Related Bacteria Dominant to Lactobacillus in Rhesus Macaque.

The high incidence of bacterial vaginosis recurrence is common after treatment with an antibiotic agent and suggests the need for new treatments to prevent this. We conducted a randomized trial to evaluate the ability of maltose gel to treat bacterial vaginosis. Eighteen female rhesus macaques were randomly assigned, in a 2:1 ratio, to receive maltose gel or placebo gel by syringe to the fornix of the vagina for five consecutive days. We used 16S rRNA sequencing data from 70 swab samples of vaginal secretions in two groups in total on days 0, 3, and 5 after medication initiation and days 3 and 5 after medication withdrawal for the study of microbiome composition. We found that, in the placebo control group, there was no significant change in the composition and abundance of vaginal microbiota during the follow-up period. In the maltose gel test group, the abundance of Lactobacillus in the vagina microbiota increased gradually with the prolongation of the treatment time on Days 3 and 5 (ANOVA p = 6.99e−5 < 0.01) but began to decrease after the withdrawal of maltose gel, which was different from that of the control group. Correspondingly, the diversity and abundance of BV-related bacteria, Fusobacterium, Parvimonas, Mobiluncus, Campylobacter, Prevotella, and Sneathia, decreased on Day 0 to Day 5 of medication and increased after drug withdrawal in the maltose gel test group. The study confirms that maltose gel can facilitate the proliferation of Lactobacillus and promote the transition of the vaginal microbiota from BV-related bacteria dominant to Lactobacillus dominant in the rhesus macaque.

Molecular Investigation of Etiologic Agents Causing Vulvovaginal Candidiasis

Background: Vulvovaginal candidiasis (VVC) is an ordinary infection caused by Candida species. Meanwhile, a shift towards non-albicans Candida (NAC) species has been detected in VVC patients. Objectives: This study aimed at molecular identification of Candida isolates, causing VVC. Methods: Vaginal secretion samples of 320 non-pregnant vaginitis patients at Shahid Akbar-Abadi Obstetrics and Gynecology Hospital in Tehran (Iran) were collected. Samples were evaluated using mycological and molecular approaches. Vaginitis isolates were analyzed with the PCR using NL1 and NL4 primers, and the D1/D2 region of the large-subunit rRNA gene was amplified and sequenced. Results: In total, 100 Candida isolates were identified from VVC and recurrent vulvovaginal candidiasis (RVVC). Candida albicans was the most frequent (51%), followed by C. glabrata (36%), C. krusei (Pichia kudriavzevii) (8%), and C. kefyr (Kluyveromyces marxianus) (5%). 51 and 49% of isolates had C. albicans and NAC, respectively. Conclusions: Candida albicans and C. glabrata were the most common agents of vulvovaginal candidiasis. NAC spp. (49%) was found as an important agent associated with VVC.

Accuracy in clinical examinations for the diagnosis of vulvovaginitis by CANDIDA SPP. and IN VITRO susceptibility to the main antifungals

Vulvovaginal candidiasis (VVC) is a common infection. This work aims to determine the positive predictive value (PPV) of the clinical diagnosis of VVC and to characterize Candida species isolated from the vaginal mucosa. This cross-sectional study was conducted from February 2016 to February 2017 at the Gynecology and Obstetrics Outpatient Clinic of the Hospital das Clínicas, in Goiânia, Goiás State, Brazil. The study included samples of vaginal secretion from 55 women who complained of vaginal discharge and itching as their main symptoms. The PPV of the clinical diagnosis of VVC was estimated in comparison to the laboratory culture method. The phenotypic methods and molecular tests were performed to identify Candida spp. In vitro susceptibility of Candida spp. isolates to fluconazole, itraconazole, clotrimazole, nystatin, and amphotericin B was determined using the broth microdilution assay. Yeast growth using the enzymes protease, phospholipase, and hemolysin was carried out in media containing respectively bovine albumin, egg yolk, and sheep erythrocytes. A PPV of 61.8% (34/55) was determined. Among the 55 vulvovaginal samples collected, we identified 36 isolates in whichC. albicans was the most common species. High resistance to fluconazole and low minimal inhibitory concentration (MIC) values for clotrimazole, nystatin and amphotericin B were observed. All isolates were proteinase and hemolysin producers, while seven strains were phospholipase negative. The clinical diagnosis of VVC presented a moderate PPV, which meant that cultures had to be conducted in the laboratory to confirm infection. The high resistance to fluconazole and itraconazole indicated the importance of the in vitro susceptibility test.KEY WORDS: Vulvovaginal candidiasis; antifungal susceptibility; enzymatic activity

Coronavirus disease 2019 among pregnant Chinese women: case series data on the safety of vaginal birth and breastfeeding.

ObjectiveTo assess whether vaginal secretions and breast milk of women with coronavirus disease 2019 (COVID‐19) contain severe acute respiratory syndrome coronavirus 2 (SARS‐CoV‐2).DesignSingle centre cohort study.SettingRenmin Hospital of Wuhan University, Wuhan, Hubei province, China.Population We studied 13 SARS‐CoV‐2‐infected pregnant women diagnosed between 31 January and 9 March 2020.MethodsWe collected clinical data, vaginal secretions, stool specimens and breast milk from SARS‐CoV‐2‐infected women during different stages of pregnancy and collected neonatal throat and anal swabs.Main outcomes and measuresWe assessed viral presence in different biosamples.ResultsOf the 13 women with COVID‐19, five were in their first trimester, three in their second trimester and five in their third trimester. Of the five women in their third trimester who gave birth, all delivered live newborns. Among these five deliveries, the primary adverse perinatal outcomes included premature delivery (n = 2) and neonatal pneumonia (n = 2). One of nine stool samples was positive; all 13 vaginal secretion samples, and five throat swabs and four anal swabs collected from neonates, were negative for the novel coronavirus. However, one of three samples of breast milk was positive by viral nucleic acid testing.ConclusionsIn this case series of 13 pregnant women with COVID‐19, we observed negative viral test results in vaginal secretion specimens, suggesting that a vaginal delivery may be a safe delivery option. However, additional research is urgently needed to examine breast milk and the potential risk for viral contamination.Tweetable abstractNew evidence for the safety of vaginal delivery and breastfeeding in pregnant women infected with SARS‐CoV‐2, positive viral result in a breast‐milk sample.

Development of a multiplex system for the identification of forensically relevant body fluids

Currently, mRNA profiling is widely investigated for forensic body fluid identification, while it is still required to advance the approach for those casework samples of limited quantity or low quality. The inclusion of circular RNAs (circRNAs) can facilitate the detection of mRNA markers in forensic body fluid identification. In this study, a multiplex assay for forensic body fluid identification (F18plex assay) was developed by incorporating 14 tissue-specific mRNA markers with circRNAs expression, 2 mRNA markers with high abundance and 2 housekeeping markers for the discrimination of the most common forensic body fluids, including blood, menstrual blood, saliva, vaginal secretion, semen and urine. The markers employed in the F18plex assay show similar specificity to previous reports. Additionally, even if all linear transcripts were completely erased, the expected markers in target biofluids could still be identified, which should help the discrimination of those aged biological stains. Results from sensitivity testing and the detection of mixtures demonstrate good sensitivity of the multiplex assay. Generally, full biomarker profiles could be obtained with ≥1 μl of blood, saliva, or semen, and ≥1 ng of total RNAs from menstrual blood, vaginal secretion, or urine samples, respectively, using this multiplex assay under the established conditions. Collectively, the newly established multiplex assay can assist in determining the biological origin of forensic stains.

Acute Miliary Tuberculosis and Pelvic Tuberculosis after In vitro Fertilization

Nearly one-fourth of the world's population is infected with Mycobacterium tuberculosis (MTB). Female genital tuberculosis (TB) is a common cause of infertility in both developing and undeveloped countries. Furthermore, assisted reproduction treatments and pregnancy potentially increase the risk of TB infection and reactivation. In this study, we present the case of a 28-year-old infertile female without a history of TB who developed an acute miliary TB and pelvic TB after in vitro fertilization–embryo transfer (IVF-ET). Elevated serum estrogen levels during controlled ovarian hyperstimulation and T-lymphocyte function inhibition during pregnancy are the risk factors for MTB infection and reactivation. In her 7th week of gestation, the patient developed fever and spontaneously aborted. Her chest computed tomography images revealed classical miliary TB. Uterine curettage tissue and vaginal secretion samples as well as Gene X-pert MTB/rifampicin (RIF) and TB-RNA test results were positive for MTB. Histological examination of the uterine curettage tissue confirmed the diagnosis of endometrial TB. Treatment with isoniazid, RIF, pyrazinamide, amikacin, and levofloxacin was selected based on the patient's diagnosis, complications, and test results. Currently, the patient is undergoing anti-TB treatment, and her condition is stable. It is important to rule out the presence of TB in infertile patients before performing IVF-ET to avoid TB dissemination during pregnancy.

Viral Shedding of COVID-19 in Pregnant Women

Background: Since there is little awareness of the consequences of coronavirus disease 2019 (COVID-19) in pregnancy, data on the safety of vaginal delivery and breastfeeding in pregnant women with COVID-19 are urgently needed. Methods: We studied COVID-19- infected pregnant women diagnosed at the Renmin Hospital of Wuhan University between January 31 and March 9, 2020, and collected clinical data, vaginal secretion, and stool specimens during different stages of pregnancy. We also collected breast milk samples, as well as throat and anal swabs from the corresponding infants to detect the presence of the virus. Results: Of the 13 women with COVID-19, 5 were in their first trimester, 3 in their second trimester, and 5 in their third trimester. Clinical manifestations observed among the 13 women included fever (n = 8), cough (n = 5), increased leukocyte count (n = 3), lymphopenia (n = 2), impaired liver function (n = 3), dyspnea (n = 1), myalgia (n = 1), and diarrhea (n = 1). Of the 5 women during their third trimester who gave birth, all delivered live newborns. Among these 5 deliveries, the primary adverse perinatal outcomes included premature delivery (n = 2) and neonatal pneumonia (n = 2). One of the 5 women followed from the first trimester experienced a biochemical pregnancy. One of 9 stool samples was positive; and all 13 vaginal secretion samples, and 5 throat swabs and 4 anal swabs collected from newborns were negative for the novel coronavirus. However, 1 of 3 samples of breast milk was positive by viral nucleic acid testing. Conclusions: In this case series of 13 pregnant women with COVID-19, we observed negative viral test results in vaginal secretion specimens, suggesting that a vaginal delivery may be a safe delivery option. However, additional research is urgently needed to examine breast milk and the potential risk for viral contamination.

Antimicrobial activity of Asteraceae species against bacterial pathogens isolated from postmenopausal women

PurposeInvestigation of the antibacterial action of aqueous extracts of Bidens sulphurea, Bidens pilosa, and Tanacetum vulgare, species of Asteraceae family that are popularly used for the treatment of genito-urinary infection.MethodsThe minimum inhibitory concentration (MIC) and minimal bacterial concentration (MBC) of the extracts against standard strains of Staphylococcus aureus (ATCC25923), Enterococcus faecalis (ATCC29212), Escherichia coli (ATCC25922), and Pseudomonas aeruginosa (ATCC27853) and against bacteria that were isolated from cultures of vaginal secretions and urine from menopausal women with a diagnosis of recurrent urinary tract infections (rUTI) were determined by broth microdilution.ResultsThe MIC values of the three extracts against Gram-positive and Gram-negative standard bacterial strains ranged from 7.81 to 125.00 mg ml-1, and the MBC values ranged from 7.81 to 500.00 mg ml-1. However, B. sulphurea was more efficient. In the urine samples, the three extracts inhibited the growth of coagulase-negative Staphylococcus spp., and the B. pilosa was the most active extract against E. coli compared with the other ones. For the vaginal secretion samples, no significant differences in the inhibition of coagulase-positive Staphylococcus spp. and P. mirabilis were found among the extracts. T. vulgare and B. sulphurea were more effective in inhibiting coagulase-negative Staphylococcus spp. compared with B. pilosa. E. coli was more susceptible to the B. sulphurea extract compared with the B. pilosa and T. vulgare extracts.ConclusionThe present results suggested the potential medicinal use of Asteraceae species, especially B. sulphurea, as therapeutic agents against rUTI-related bacteria.

Genotyping of different clinical Candida albicans isolates using RAPD Analysis

Random Amplification of Polymorphic DNA (RAPD–PCR) is a powerful discriminative assay comparing several clinical isolates by amplifying the target DNA genome by a group of short random primers. In clinical mycology the infections with Candida species increase, especially Candida albicans and their complications become a serious problem, So this work aims to determine the genetic typing of clinical C. Albicans isolated from ICU patient's (urine, blood, vaginal secretions, and bronchoalveolar lavage) who existing in Elarreish hospital, Egypt based on RAPD-PCR assay. To investigate their genetic relationships. Totally one hundred samples of 40 samples of urine, 30 samples of blood, 20 samples of vaginal secretions and 10 samples of Bronchoalveolar lavage were collected. About 50 different species of Candida were isolated, 28 species out of them were identified by standard-taxonomic criteria and molecular technique as Candida albicans (n =28). Candida albicans represents 14 samples of urine (35%), 7 samples of blood (23%), 3 samples of vaginal secretions (10%), and 4 samples of bronchoalveolar lavage (40%). Genetic relevance of the different 28 Candida albicans was analyzed by (RAPD) assay using 6 different primers OPA1, OPA2, OPA3, OPA7, OPA8, and OPA9. Only three primers OPA3, OPA7, and OPA9 allow discrimination among the Candida albicans isolates. PCR bands were scored as present or absent and the genetic similarity between them was determined using Jaccard's coefficient(Sj). Dendrograms constructed using the UPGMA method showed that 12, 14, and 20 different genotypes can be identified by OPA3, OPA7, and OPA9 primers respectively since they showed a high degree of genetic heterogeneity. Data indicated that RAPD analysis is useful for providing genotypic characteristics for C. albicans in the epidemiological investigation.

Differences in the Composition of Vaginal Microbiota between Women Exhibiting Spleen-Deficiency Syndrome and Women with Damp-Heat Syndrome, Two of the Most Common Syndromes of Vaginitis in Traditional Chinese Medicine.

Spleen-deficiency syndrome and damp-heat syndrome are the two most common syndromes of vaginitis in traditional Chinese medicine (TCM). Although it is known that the vaginal microbiota is closely associated with vaginitis, present studies have not fully elucidated the relationship between the composition of the vaginal microbiome and type of TCM syndrome because of the limitations in the present reductionist approaches. Samples of vaginal secretions were collected from patients with bacterial vaginitis and healthy subjects with spleen-deficiency syndrome and damp-heat syndrome, in order to analyze the constitution of the vaginal microflora using 16S rRNA sequencing methods that encompass taxonomic units, alpha diversity rarefaction curves, and principal component analyses. This prospective study indicated that there was a statistically significant difference in the composition of the vaginal microbiome between patients with spleen-deficiency syndrome and patients with damp-heat syndrome. Streptococcus was the dominant microbiota in patients with spleen-deficiency syndrome. This can serve as a biomarker for differentiating spleen-deficiency syndrome and damp-heat syndrome. In addition, as indicated by the findings on the samples, patients with bacterial vaginitis of dominant abundance in Pseudomonadaceae might be prone to manifest spleen-deficiency syndrome, while patients with bacterial vaginitis of dominant abundance in Prevotella might be prone to manifest damp-heat syndrome. These present findings can provide a new approach to acquire a scientific understanding of the syndromes of TCM, which in turn would benefit the development of personalized medicine, in terms of ancient medicine and complex biological systems.

The Application of Realtime Fluorescence Quantitative PCR for Prenatal Screening of Group B Streptococcal Infections

Objective: In the prenatal screening, several different methods were used to detect the presence of group B streptococcus (GBS) infection, in this assay, the diagnostic value and clinical signifcance of the application of realtime fluorescent PCR were explored. Methods: A total of 86 women with 35-37 weeks pregnancy were enrolled, vaginal secretion samples were collected. Fluorescence PCR, bacterial culture and gene sequencing were used to detect whether there was GBS infection, and the results obtained were compared and analyzed. Results: 10 subjects were detected to be positive for GBS by fluorescence PCR (the positive rate was 11.6%), however, only 4 cases were positive for GBS by bacterial culture method (the positive rate was 4.7%). There was a statistically signifcant difference in the positive rate between the two methods (P&lt;0.01). Compared with the results of gene sequencing, the detection of GBS infection by fluorescence PCR has an accuracy of 95.2%, and the sensitivity was 90.9% with 100% specificity. Conclusion: The application of realtime fluorescence quantitative PCR for the detection of GBS infection is signifcantly better than the use of bacterial culture method. Compared with the gold standard method (gene sequencing method), its detection effciency, accuracy, sensitivity and specifcity are relatively high. In summary, PCR for prenatal screening of GBS is worthy of promotion in clinical practice.