BACKGROUND: Next-generation sequencing (NGS) can detect many more microorganisms of a microbiome than traditional methods. This study aimed to analyze the vaginal microbiomes of Korean women by using NGS that included bacteria and other microorganisms. The NGS results were compared with the results of other assays, and NGS was evaluated for its feasibility for predicting vaginitis. METHODS: In total, 89 vaginal swab specimens were collected. Microscopic examinations of Gram staining and microbiological cultures were conducted on 67 specimens. NGS was performed with GS junior system on all of the vaginal specimens for the 16S rRNA, internal transcribed spacer (ITS), and Tvk genes to detect bacteria, fungi, and Trichomonas vaginalis. In addition, DNA probe assays of the Candida spp., Gardnerella vaginalis, and Trichomonas vaginalis were performed. Various predictors of diversity that were obtained from the NGS data were analyzed to predict vaginitis. RESULTS: ITS sequences were obtained in most of the specimens (56.2%). The compositions of the intermediate and vaginitis Nugent score groups were similar to each other but differed from the composition of the normal score group. The fraction of the Lactobacillus spp. showed the highest area under the curve value (0.8559) in ROC curve analysis. The NGS and DNA probe assay results showed good agreement (range, 86.2-89.7%). CONCLUSIONS: Fungi as well as bacteria should be considered for the investigation of vaginal microbiome. The intermediate and vaginitis Nugent score groups were indistinguishable in NGS. NGS is a promising diagnostic tool of the vaginal microbiome and vaginitis, although some problems need to be resolved.
Area Under Curve ; Bacteria/*genetics/isolation & purification ; Bacterial Proteins/genetics ; Candida/*genetics/isolation & purification ; Female ; Fungal Proteins/genetics ; Gardnerella vaginalis/genetics/isolation & purification ; High-Throughput Nucleotide Sequencing ; Humans ; *Microbiota ; RNA, Ribosomal, 16S/chemistry/genetics/metabolism ; ROC Curve ; Sequence Analysis, DNA ; Trichomonas vaginalis/genetics/isolation & purification ; Vagina/*microbiology ; Vaginitis/*diagnosis/microbiology

Trichomonas vaginalis induces proinflammation in cervicovaginal mucosal epithelium. To investigate the signaling pathways in TNF-alpha production in cervical mucosal epithelium after T. vaginalis infection, the phosphorylation of PI3K/AKT and MAPK pathways were evaluated in T. vaginalis-infected SiHa cells in the presence and absence of specific inhibitors. T. vaginalis increased TNF-alpha production in SiHa cells, in a parasite burden-dependent and incubation time-dependent manner. In T. vaginalis-infected SiHa cells, AKT, ERK1/2, p38 MAPK, and JNK were phosphorylated from 1 hr after infection; however, the phosphorylation patterns were different from each other. After pretreatment with inhibitors of the PI3K/AKT and MAPK pathways, TNF-alpha production was significantly decreased compared to the control; however, TNF-alpha reduction patterns were different depending on the type of PI3K/MAPK inhibitors. TNF-alpha production was reduced in a dose-dependent manner by treatment with wortmannin and PD98059, whereas it was increased by SP600125. These data suggested that PI3K/AKT and MAPK signaling pathways are important in regulation of TNF-alpha production in cervical mucosal epithelial SiHa cells. However, activation patterns of each pathway were different from the types of PI3K/MAPK pathways.
Cell Line ; Cervix Uteri/enzymology/metabolism/*parasitology ; Epithelial Cells/*enzymology/metabolism/parasitology ; Female ; Humans ; *MAP Kinase Signaling System ; Mucous Membrane/*enzymology/metabolism/parasitology ; Phosphatidylinositol 3-Kinases/genetics/*metabolism ; Proto-Oncogene Proteins c-akt/genetics/*metabolism ; Trichomonas Vaginitis/*enzymology/genetics/metabolism/parasitology ; Trichomonas vaginalis/*physiology ; Tumor Necrosis Factor-alpha/genetics/*metabolism

BACKGROUND: The female genital tract is equipped to deal with a variety of foreign substances including a wide array of microorganisms. It is important to consider Candida-bacterial interactions in balance between healthy colonization versus vaginitis. The objectives of this study were to evaluate the association between microorganism distribution and vaginitis, and to investigate the possibility of an interaction between vaginal Candida and other microorganisms in female genital tract. METHODS: A total of 516 vaginal secretions were collected between October 2008 and June 2010 from patients with suspected vaginitis. Identification of Candida species and detection of 6 fastidious microorganisms (Trichomonas vaginalis, Neisseria gonorrhoeae, Chlamydia trachomatis, Mycoplasma hominis, Mycoplasma genitalium, and Ureaplasma urealyticum) were performed using a VITEK 2 system (bioMerieux, Inc., Hazelwood, MO, USA) and multiplex PCR (Seegene, Biotechnology, Inc., Seoul, Korea), respectively. RESULTS: M. genitalium, U. urealyticum, and C. trachomatis were more often detected in association with vaginal candidiasis. A statistically significant association between Candida and M. genitalium was observed (P<0.05). N. gonorrhoeae was detected less often in women with vaginal candidiasis. CONCLUSION: The results of this study suggest the possibility that vaginal Candida may associate with some microorganisms in patients with vaginitis. Further studies will be required to define the Candida-bacterial interactions and its mechanisms.
Bacteria ; Biotechnology ; Candida ; Candidiasis ; Chlamydia trachomatis ; Colon ; Female ; Humans ; Microbial Interactions ; Multiplex Polymerase Chain Reaction ; Mycoplasma genitalium ; Mycoplasma hominis ; Neisseria gonorrhoeae ; Trichomonas vaginalis ; Ureaplasma ; Vaginitis

BACKGROUND: Infectious vaginitis is caused primarily by three different groups of microbial pathogens (Trichomonas vaginalis, Candida spp., and Gardnerella vaginalis). The objective of this study was to compare the Affirm VPIII assay using a DNA hybridization technique with the Papanicolaou (Pap) smear test and the Gram stain in the detection and identification of these three organisms. METHODS: A total of 300 vaginal samples were collected from women that were either symptomatic for vaginitis or asymptomatic women that were being seen for routine obstetric or gynecological care. The presence of T. vaginalis, Candida spp., and G. vaginalis was evaluated by using the Affirm VIII assay (Becton Dickinson, USA), Pap smear test, and Gram stain method, respectively. RESULTS: With the Affirm VPIII assay, 1 (0.3%) patient tested positive for T. vaginalis, 99 (33.0%) patients were positive for G. vaginalis, and 18 (6.0%) were positive for Candida spp. The detection rates of Trichomonas infection, bacterial vaginosis and candidiasis by the Pap smear test and Gram stain method were 0.7% versus 0%, 16.3% versus 35.7%, and 1.7% versus 9.7%, respectively. The differences between the detection rates of the above three organisms between the Pap smear test and the Gram stain method were statistically significant (p<0.05). CONCLUSION: The Affirm VPIII assay was more sensitive than the Pap smear test and more specific than the Gram stain method for the detection and identification of these three organisms. In addition, the results of the Affirm VPIII assay are quick to obtain and are simple and easy to interpret.
Candida ; Candidiasis ; Chimera ; DNA ; Female ; Gardnerella ; Gardnerella vaginalis ; Humans ; Trichomonas ; Trichomonas Infections ; Trichomonas vaginalis ; Vaginal Smears ; Vaginitis ; Vaginosis, Bacterial

PURPOSE: We aimed to investigate the detection of nanobacteria (NB) from expressed prostatic secretions (EPS) in patients with category III chronic prostatitis/chronic pelvic pain syndrome (CP/CPPS) and from vaginal swabs in patients with vaginitis by reverse transcriptase polymerase chain reaction (RT-PCR) and to evaluate the association between NB and Neisseria gonorrhea, Chlamydia trachomatis, Ureaplasma urealyticum (U. urealyticum), Mycoplasma hominis, Trichomonas vaginalis, and Mycoplasma genitalium. MATERIALS AND METHODS: A group of 11 men attending a specialized CP/CPPS clinic and a group of 157 women who reported symptoms of lower genital tract infection were enrolled in this study. NB were detected by RT-PCR. A Seeplex Sexually Transmitted Disease Detection assay (Seegene Inc., Seoul, Korea) was used that could detect DNA for 6 types of sexually transmitted pathogens. RESULTS: In EPS samples, the detection rate of NB in patients with CP/CPPS was 9.1%, and 9 (5.7%) of 157 vaginitis patients showed positive results in RT-PCR for NB in vaginal swabs. Associations observed among the 7 microorganisms included 6 (54.5%) patients who tested positive on EPS and 75 (47.8%) patients who tested positive on vaginal swabs. Five patients with vaginitis were found to have monoinfection of NB (6.7%). CONCLUSIONS: We found that conventional RT-PCR for NB was rapid, simple, low in cost, and easily available for the detection of NB, and that NB may be a possible etiological factor for vaginitis and CP/CPPS. The prevalence of U. urealyticum among the four patients with NB coinfection was 75%; the presence of U. urealyticum might therefore raise suspicion for nanobacterial infection.
Calcifying Nanoparticles ; Chlamydia trachomatis ; Coinfection ; DNA ; Female ; Gonorrhea ; Humans ; Male ; Mycoplasma ; Mycoplasma hominis ; Nanoparticles ; Neisseria ; Pelvic Pain ; Prevalence ; Prostatitis ; Reproductive Tract Infections ; Reverse Transcriptase Polymerase Chain Reaction ; RNA-Directed DNA Polymerase ; Sexually Transmitted Diseases ; Trichomonas vaginalis ; Ureaplasma urealyticum ; Vaginitis

BACKGROUND: Bacterial vaginitis (BV) and Trichomonas vaginitis are the most frequently recurring infectious diseases in women. Therefore, accurate tests for post-treatment follow-up are required. A multiplex PCR assay allows for the simultaneous detection of multiple pathogens in a single specimen. In this study, we assessed the clinical implications of multiplex PCR detection of fastidious microorganisms causing vaginitis. METHODS: A total of 216 vaginitis patients who presented to Chung-Ang University Yongsan Hospital with more than one positive result on multiplex PCR (Trichomonas vaginalis (TV), Neisseria gonorrhoeae (NG), Chlamydia trachomatis (CT), Ureaplasma urealyticum (UU), Mycoplasma genitalium (MG), Mycoplasma hominis (MH)) were retrospectively enrolled in this study. Each patient's clinical symptoms, initial treatment and follow-up for BV, and other related test results were also retrospectively reviewed. RESULTS: The most commonly reported symptom was abnormal discharge, followed by pruritis (73.1%), lower abdominal pain (38.4%), urination difficulties (13%), and others such as fever. According to the multiplex PCR results, there were 116 cases (35.8%) of MH, 86 cases (26.5%) of UU, 62 cases (19.1%) of CT, and 84 cases (38.9%) were mixed infections. Among those patients with single infections, treatment changed for 63 cases (65.6%) while treatment remained unchanged for 17 (17.7%) after PCR results were reported. CONCLUSION: The diagnosis of BV using multiplex PCR is clinically effective and the results of which can be incorporated in antibiotic selection for patients with multiple sexually transmitted diseases (STD). Multiplex PCR may be especially helpful in the diagnosis of patients in whom the differentiation of STD pathogens is difficult using traditional methods.
Abdominal Pain ; Chlamydia trachomatis ; Coinfection ; Communicable Diseases ; Female ; Fever ; Follow-Up Studies ; Humans ; Multiplex Polymerase Chain Reaction ; Mycoplasma genitalium ; Mycoplasma hominis ; Neisseria gonorrhoeae ; Polymerase Chain Reaction ; Pruritus ; Retrospective Studies ; Sexually Transmitted Diseases ; Trichomonas Vaginitis ; Ureaplasma urealyticum ; Urination ; Vaginitis ; Vaginosis, Bacterial

Trichomonas vaginitis is a sexually transmitted disease by Trichomonas vaginalis infection especially in adults that is significantly associated with preterm delivery, premature rupture of membrane and low-birth-weight neonates. Although neonatal infection is rare, a vertical transmission might substantially occur by direct contact of protozoal infection. The reported clinical manifestation of trichomoniasis is purulent vaginal discharge in only severely infected neonates. I experienced a case of virulent neonatal Trichomonas vaginitis which was confirmed by vaginal wet mount examination in a neonate born full-term, complaining of excessive purulent vaginal discharge, with no perinatal complication of premature rupture of membrane or low-birth-weight. The patient had complete resolution of symptom after the administration of metronidazole, thus I report this case along with a brief review of literatures.
Adult ; Humans ; Infant, Newborn ; Membranes ; Metronidazole ; Rupture ; Sexually Transmitted Diseases ; Trichomonas vaginalis ; Trichomonas Vaginitis* ; Trichomonas* ; Vaginal Discharge

Reactive arthritis occurs after a preceding infection such as urogenital or gastroenteral tract infection. Trichomoniasis, due to Trichomonas vasinalis infection, is one of the most common causes of vaginitis. Reactive arthritis associated with trichomoniasis is uncommon and there has been no report in Korea. We present a 28 year-old woman who had oligoarthritis after Trichomonas vaginalis infection. The arthritis subsided with treatment of the Trichomoniasis with metronidazole and non-steroidal anti-inflammatory drug.
Adult ; Arthritis ; Arthritis, Reactive* ; Female ; Humans ; Korea ; Metronidazole ; Trichomonas Infections ; Trichomonas vaginalis* ; Trichomonas* ; Vaginitis

Vaginal trichomoniasis, caused by Trichomonas vaginalis, is the most common sexually transmitted disease. More than 170 million people worldwide are annually infected by this protozoan. In the Republic of Korea, 10.4% of women complaining of vaginal symptoms and signs were found to be infected with T. vaginalis. However, despite its high prevalence, the pathogenesis of T. vaginalis infection has not been clearly characterized although neutrophil infiltration is considered to be primarily responsible for the cytologic changes associated with this infection. We hypothesized that trichomonads in the vagina sometime after an acute infection secrete proteins like excretorysecretory product that have a chemotactic effect on neutrophils, and that these neutrophils are further stimulated by T. vaginalis to produce chemokines like IL-8 and GRO-alpha, which further promote neutrophil recruitment and chemotaxis. Thus, neutrophil accumulation is believed to maintain or aggravate inflammation. However, enhanced neutrophil apoptosis induced by live T. vaginalis could contribute to resolution of inflammation. Macrophages may constitute an important component of host defense against T. vaginalis infection. For example, mouse macrophages alone and those activated by lymphokines or nitric oxide are known to be involved in the extracellular killing of T. vaginalis. In the host, T. vaginalis uses a capping phenomenon to cleave host immunoglobulins with proteinases and thus escape from host immune responses. Recently, we developed a highly sensitive and specific diagnostic polymerase chain reaction (PCR) technique using primers based on a repetitive sequence cloned from T. vaginalis (TV-E650), and found that the method enables the detection of T. vaginalis at concentrations as low as 1 cell per PCR mixture.
Trichomonas vaginalis/cytology/*isolation & purification/pathogenicity ; Trichomonas Vaginitis/drug therapy/*epidemiology/parasitology ; Korea/epidemiology ; Humans ; Female ; Animals

Three common vaginal infections are bacterial vaginosis (BV), vulvovaginal candidiasis (VVC), and trichomonas vaginitis (TV). However their symptoms and signs are similar and atypical and other non-infectious vaginal inflammations will be the causes of identical symptoms. Hence accurate diagnosis is the first step in management of vaginitis and then appropriate therapy should be conducted. Metronidazole is the drug of choice for the treatment of BV and TV. Topical or oral azole antifungal regimens are highly effective in reliving symptoms of VVC. Physicians who treat the woman having vaginitis should consider the fact that the disease may be chronic, frequent and recurrent due to antibiotics abuse and over the counter antimycotic agents. Also predisposing factors must be removed before medical therapy.
Anti-Bacterial Agents ; Candidiasis, Vulvovaginal ; Causality ; Danazol ; Diagnosis ; Female ; Humans ; Inflammation ; Metronidazole ; Trichomonas Vaginitis ; Vaginitis* ; Vaginosis, Bacterial