Cost analysis of four types of surgeries for pelvic organ prolapse in a Japanese population.

INTRODUCTION AND HYPOTHESIS: To compare the perioperative costs analysis between laparoscopic/transvaginal and the mesh/non-mesh surgeries for pelvic organ prolapse (POP) in Japan. MATERIALS AND METHODS: From April 2013 to April 2017, 890 patients who underwent POP surgeries were enrolled in this study. Regarding transvaginal native tissue repair (TV-NTR: transvaginal hysterectomy with colpocleisis), transvaginal mesh surgery (TVM), laparoscopic native tissue repair (L-NTR: laparoscopic hysterectomy and uterosacral ligament colposuspension), and laparoscopic sacrocolpopexy (LSC), a retrospective observational study was performed. Patients' age, operation time, blood loss, perioperative complications, length of hospital stay, pre-/postoperative quality of life (QOL) scores, were reviewed from the medical records. The net income, which was calculated by using the income (the operation/anesthesia fee) and the costs (the labor and consumables costs for operation/anesthesia), was evaluated. RESULTS: The operation fees of the L-NTR ($4250) and the LSC ($4833) groups were higher than that of the TV-NTR ($2652) and the TVM ($2913) groups. The labor costs and consumables costs of operation were higher in the LSC ($1589) and the L-NTR ($1500) groups than the TV-NTR ($180) and the TVM ($178) groups. The consumables costs for anesthesia in the four groups were equal. The operation hours were significantly shorter in the TV-NTR and the TVM groups than the L-NTR and the LSC groups. CONCLUSIONS: We found that TVM operation was an economically excellent and the most efficient POP operation with shorter operation time and less consumables.

Guideline for Gynecological Practice in Japan: Japan Society of Obstetrics and Gynecology and Japan Association of Obstetricians and Gynecologists 2020 edition.

Nine years after the first edition of The Guideline for Gynecological Practice, which was jointly edited by The Japan Society of Obstetrics and Gynecology and The Japan Association of Obstetricians and Gynecologists, the 4th Revised Edition was published in 2020. The 2020 Guidelines includes 4 additional clinical questions (CQ), which brings the total to 99 CQ (12 on infectious disease, 29 on oncology and benign tumors, 29 on endocrinology and infertility and 29 on healthcare for women). Currently, a consensus has been reached on the Guidelines, and therefore, the objective of this report is to present the general policies regarding diagnostic and treatment methods used in standard gynecological outpatient care that are considered appropriate. At the end of each answer, the corresponding Recommendation Level (A, B, C) is indicated.

In vitro characteristics of intravaginal Lactobacilli; why is L. iners detected in abnormal vaginal microbial flora?

PURPOSE: Our purpose is to investigate the reasons why Lactobacillus iners is detected in abnormal vaginal microbial flora. METHODS: In this study, in vitro characteristics of four type strains (L. crispatus, L. iners, L. gasseri, and L. jensenii) were examined by measuring the growth speed by OD660, and acid resistance, with gram stain and Live/Dead stain. RESULTS: The growth speed was L. gasseri > L. jensenii > L. crispatus > L. iners. Bacterial counts of all Lactobacilli in MRS medium began to decrease at the middle of the log-phase of the growth curve. In addition, L. iners grew to 106 CFU/mL and the others grew to 108 CFU/mL. L. iners was mostly Gram-negative with very short rod, while the others were mostly Gram-positive rods. L. iners was completely killed in the pH 3 medium, however, the others grew (in pH 3 medium) in 1/100 order compared with those in the pH 6 medium. CONCLUSION: L. iners was not a typical gram-positive long rod Lactobacilli and presented weak acid-resistance. The reasons why L. iners is detected in abnormal vaginal microbial flora were presumed to be due to the unique morphologic and microbiologic characteristics.

Changes of intravaginal microbiota and inflammation after self-replacement ring pessary therapy compared to continuous ring pessary usage for pelvic organ prolapse.

AIM: Continuous usage of a ring pessary for pelvic organ prolapse may cause a disturbance of intravaginal microbiota and intravaginal mucosal damage. To avoid the side effects of continuous ring pessary therapy, daily self-replacement of the ring pessary is recommended. The purpose of this study is to evaluate the outcomes of self-replacement versus continuous ring usage, by analysing clinical findings and intravaginal microbiota. METHODS: Thirty seven patients who managed self-replacement ring pessary therapy and 13 patients with continuous ring pessary therapy participated in this study. The clinical symptoms were checked at 1 month after the initial ring pessary insertion. The changes in the intravaginal microbiota were evaluated by conventional methods, i.e. pH in the vagina, Lactobacillary grade, Nugent score, inflammatory cell counts, and culture-based bacterial detection methods. In addition, our clone library method using 16S rRNA sequencing of vaginal fluid was performed. RESULTS: Patients were divided into four groups: self-replacement pre/post-menopause and continuous and pre/post-menopause. Five patients of the self-replacement group (n = 37) and all patients of the continuous use group (n = 13) complained of increased discharge. However, both the conventional methods and the clone library method revealed that the number of the self-replacement group patients who had abnormal intravaginal microbiota were not significantly different from that of the continuous use group. CONCLUSION: Daily self-replacement ring pessary therapy prevented adverse clinical symptoms. However, abnormal intravaginal microbiota was frequently observed during self-replacement of ring pessary therapy as with continuous usage. Regardless, pelvic examinations should be performed routinely.

Laparoscopic uterosacral ligament colpopexy for apical support in addition to hysterectomy for pelvic organ prolapse.

AIM: To evaluate the perioperative complications and the anatomical outcomes of our laparoscopic uterosacral ligament (USL) colpopexy, which is a novel laparoscopic technique for the management of uterine prolapse. The objective was to report on outcome after 2 years of a technique using laparoscopic USL colpopexy. METHODS: A total of 152 uterine prolapse patients underwent laparoscopic USL colpopexy from May 2013 to April 2015. We described the surgical technique and performed a retrospective analysis of this laparoscopic technique. Patients underwent standardized assessment and examination using pelvic organ prolapse quantification (POP-Q) score. The dependent values of Ba point (bladder), C point (vaginal cuff) and Bp point (rectum) were recorded preoperatively, and at 1, 3, 6, 12 and 24 months of postoperative examination. Pre/postoperative data were compared using the Kaplan-Meier method. RESULTS: Mean age, operative time and amount of hemorrhage were 68.2 ± 7.5 years, 118.3 ± 36.4 min and 60.5 ± 73.3 mL, respectively. Overall recurrent prolapse, which was defined as POP-Q stage II or higher, was noted in 29 patients (19%). However, only 2 patients presented recurrent rectocele among 51 patients with preoperative POP-Q stage II of uterine prolapse alone (recurrence rate; 3.9%). The pre/postoperative average POP-Q scores were -0.2/-2.7 cm (P < 0.05) at Ba point, -1.9/-5.1 cm (P < 0.05) at C point and -2.4/-2.3 cm (P = 0.06) at Bp point. CONCLUSION: Laparoscopic visualization of uterosacral ligaments may result in safe colpopexy. Our results show this will be a useful procedure for apical support as native tissue repair.

Aiming for complete safety of first trocar insertion of laparoscopic surgery: usefulness of preoperative ultrasonography of umbilical region.

AIM: Vascular or intestinal injuries at the time of the first trocar insertion can cause serious complications during laparoscopic surgeries. In this study, we evaluate the usefulness of ultrasound scans of the umbilical region as well as intraumbilical conditions to help prevent serious complications. METHODS: The subjects included 430 cases who underwent laparoscopic gynecologic surgeries. The umbilical ultrasound scan was performed after tracheal intubation to observe the intestinal movements associated with respiration. Structures of the umbilical region as well as peristalsis and the movement of the intestinal tract were observed. Then, the thickness of the subcutaneous fat (between the umbilical skin surface and the rectus fascia) and the preperitoneal fat (between the rectus fascia and the peritoneum) were measured. The relationship between body mass index (BMI), insertion time of the first trocar and ultrasound measurements were analyzed. RESULTS: The anatomical structures of the umbilical region (the subcutaneous tissue and the preperitoneal fat) were clearly observed in all cases. The BMI score had a significant relationship with subcutaneous fat thickness (r = 0.547), but remarkably not with preperitoneal fat thickness (r = 0.174). There was no significant relationship between BMI and insertion time. However, insertion time of the first trocar had a significant relationship with preperitoneal fat thickness (r = 0.534). CONCLUSIONS: Preoperative ultrasonography of the umbilical region is asimple process, and it is helpful inpreventing serious complications caused by the first trocar insertion. We have found that preperitoneal fat thickness seems to be an important factor in predicting the potential difficulty of the first trocar insertion.

Pharmacokinetic and Pharmacodynamic Markers of Mycophenolic Acid Associated with Effective Prophylaxis for Acute Graft-Versus-Host Disease and Neutrophil Engraftment in Cord Blood Transplant Patients.

Mycophenolate mofetil (MMF) is a prodrug of mycophenolic acid (MPA) and is frequently used to prevent acute graft-versus-host disease (aGVHD) in patients receiving hematopoietic stem cell transplantation (HSCT). However, optimal doses of MMF and target MPA concentrations in HSCT patients have not been defined. In this study, relationships between pharmacokinetic or pharmacodynamic markers of MPA and successful aGVHD prevention and neutrophil engraftment were evaluated to inform individualized MPA treatments in HSCT patients. We recruited 35 patients undergoing cord blood transplantation (CBT) who were treated with MMF. Area under the concentration-time curves from 0 to 24 hours (AUC0-24) for free MPA and MPA acyl glucuronide (AcMPAG) at 1 week after the start of MMF treatments were significantly higher in patients with gastrointestinal (GI) aGVHD at stage ≥I than those at stage 0. Patients with faster neutrophil engraftment had higher free MPA AUC0-24 at 1 week after the start of MMF treatments compared with those with slower neutrophil engraftment. Inosine-5'-monophosphate dehydrogenase activity in peripheral blood mononuclear cells and single nucleotide polymorphisms in genes that were previously associated with MPA pharmacokinetics and pharmacodynamics were not an independent predictor for the clinical outcomes. Receiver-operating characteristic model analyses showed that cutoff values of AUC0-24 for successful GI aGVHD prevention were .689 and 15.6 µg⋅hour⋅mL-1 for free MPA and AcMPAG, respectively. In addition, the cut-off value of free MPA AUC0-24 for neutrophil engraftment by day 25 was .405 µg⋅hour⋅mL-1. In conclusion, free MPA AUC0-24 may be a better predictor of the prevention of GI aGVHD and neutrophil engraftment compared with that of total MPA in patients receiving CBT. Hence, monitoring of the free MPA AUC0-24 between .405 and .689 µg⋅hour⋅mL-1 could be considered informative of individualized MPA treatments in CBT patients.

Novel technique for adhesion barrier insertion by using cut-off surgical glove fingers in laparoscopic surgery.

AIM: Laparoscopic surgery causes fewer postoperative adhesions than laparotomy and adhesion barriers are used to lessen the chance of adhesion. Sodium hyaluronate and carboxymethylcellulose (SH-CMC), a bioresorbable membrane, is effective in preventing adhesions. However, its properties make it difficult to insert the film into the abdominal cavity and to place it into the target area during laparoscopic surgeries. Here we report a novel technique for applying SH-CMC during laparoscopic surgeries by using cut-off surgical glove fingers. METHODS: In our dry-box experiment, we found that SH-CMC became more flexible after moisturizing for 30 s. Therefore, for this procedure, we moisturizing both sides of the SH-CMC sheet (7.35 × 6.35 cm) on wet gauze (for 30 s). The film was rolled into cut-off surgical glove fingers and inserted into the abdominal cavity through a 12-mm trocar. The film was then removed from the glove finger and unrolled onto the target area (glove-finger method). RESULTS: The dry-box experiment revealed that moisturizing both sides of SH-CMC for 30 s/side, 15 min after package removal, made the film more flexible. Ninety of 96 attempts with SH-CMC were successful among the 24 patients who underwent total laparoscopic hysterectomy with our glove-finger method. CONCLUSION: Our glove-finger method can realize the smooth application of SH-CMC to the target area during laparoscopic surgeries.

How to reduce the operative time of laparoscopic sacrocolpopexy?

OBJECTIVE: Laparoscopic sacrocolpopexy (LSC) has been reported to achieve lower recurrence rates, shorter recovery time, and less dyspareunia. However, as a pelvic organ prolapse (POP) surgery, LSC is problematic because it requires specific techniques and it takes a comparatively longer operative time. In this study, we present our surgical techniques of LSC and their effectiveness for shortening operative times and raising safety. METHODS: Thirty-four women with stage 2 or greater POP who underwent LSC in our hospital between September 2014 and October 2015 were enrolled in this study. The notable points of our operative procedures are as follows: (1) fixing the sigmoid colon to the left lateral abdominal wall for a clearer visualization of the sacral promontory, (2) making a retroperitoneal tunnel (not opening the peritoneum) from the sacral promontory to the Douglas pouch, (3) dissection of the vaginal wall after transvaginal hydrodissection, (4) fixation of mesh to the vaginal wall by using absorbable tacks, and (5) limiting usage of posterior mesh for the patients with posterior vaginal wall descent. RESULTS: The median operative time was 140 (range, 90-255) minutes, and blood loss was 50 (range, 10-1600) mL. The operative time decreased as the surgical techniques improved through experience. No major intra- or postoperative complications occurred. The mean follow-up period was 4 (range, 1 -14) months, and only one patient presented a recurrent grade 2 cystocele. CONCLUSION: Our unique procedures will help shorten operative times and reduce complications of LSC.

Laparoscopic fixation of the vaginal cuff to the uterosacral ligaments at the time of hysterectomy.

INTRODUCTION AND HYPOTHESIS: Transvaginal ipsilateral uterosacral ligament colpopexy for pelvic organ prolapse (POP), which was reported by Shull et al. (Shull's colpopexy) in 2000, is one of the most frequently performed non-mesh pelvic floor reconstructive surgical procedures. Despite its excellent anatomical outcomes, ureteral injury and difficulty in uterosacral ligament detection (especially in patients with severe POP) are typical issues with this procedure. METHOD: This video demonstrates the procedure in a 58-year-old woman, gravida 2 para 2, with POP-Q stage II uterine prolapse and stage I cystocele. At the beginning of the operation, the bilateral uterosacral ligaments were confirmed and separated from the ureters after entering the retroperitoneal space. The inferior hypogastric nerve and pelvic nerve plexus were also separated from the uterosacral ligament. After total laparoscopic hysterectomy, three ipsilateral delayed absorbable monofilament sutures were placed between the uterosacral ligament and the vaginal cuff. Retroperitonealization was then performed using a continuous suture with closure of the Douglas pouch. CONCLUSION: Laparoscopic Shull's colpopexy for POP is a secure procedure with the advantages of laparoscopy (magnification and sharing the operative field). This may become one of the most useful operations for apical support as native tissue repair.

Objective diagnosis of arrested labor on transperineal ultrasound.

AIM: Recent developments in transperineal ultrasound imaging of the pelvis have prompted trials to objectively evaluate labor progression for labor management. We evaluated the accuracy of transperineal ultrasound in diagnosing arrest of labor. METHODS: Transperineal ultrasound and digital pelvic examinations were performed simultaneously in 63 term laboring patients (singleton fetuses in cephalic presentation). We analyzed a total of 216 ultrasound images (Sonography Volume Computer Aided Display Labor [Sono VCAD Labor®] installed in Voluson E8 ultrasound). We examined the correlation between the three ultrasound parameters head direction (HD), progression distance (PD), and progression angle (PA), and digital pelvic examination findings during labor in a transvaginal delivery group and an arrested labor group. RESULTS: The coefficient of correlations between HD/PD/PA and cervical dilation/fetal station were 0.667/0.657/0.706 and 0.667/0.751/0.803, respectively. The three parameters had strong correlations with digital pelvic examination (P < 0.05). In the 11 cases (17%) of cesarean section due to arrested labor, the position of the fetal head was visually unchanged on sequential ultrasound images. According to receiver operating characteristic curves, the significant cut-offs for HD, PD, and PA for arrested labor were 105° (P = 0.048), 35 mm (P = 0.048), and 120° (P = 0.001), respectively. CONCLUSIONS: Transperineal ultrasound imaging is helpful for objective evaluation of labor progression and the diagnosis of arrested labor.

Effects of pelvic organ prolapse ring pessary therapy on intravaginal microbial flora.

INTRODUCTION AND HYPOTHESIS: The purpose of this paper is to report our evaluation of changes in intravaginal microbial flora after ring pessary therapy for pelvic organ prolapse (POP) using conventional and our clone library method. METHODS: Thirteen patients with POP who were fitted with a ring pessary participated in this longitudinal study that incorporates data from before and 1 month after beginning ring pessary therapy. Changes in intravaginal microbial flora were evaluated by conventional methods, i.e., vaginal pH, lactobacillary grade (LAC grade), Nugent score, and culture-based bacterial detection methods. In addition, we performed our clone library method using 16S ribosomal RNA (rRNA) sequencing of vaginal fluid. RESULTS: Conventional methods revealed that most patients had abnormal intravaginal microbial flora. Mean numbers of detected bacterial species by the culture-based and our clone library method were 3.1 (1-6) and 11.8 (1-25), respectively. Our clone library method showed that Lactobacillus spp. increased in four and decreased in two cases after ring pessary therapy but reappeared after therapy in two cases; no Lactobacillus spp. were detected in five cases. CONCLUSIONS: Our study showed that ring pessary therapy did not always disturb intravaginal microbial flora, especially for patients with Lactobacillus spp. prior to ring pessary insertion. Anaerobic circumstances in the vagina after therapy seem to have induced the growth of anaerobic bacteria.

Pharmacokinetics and pharmacodynamics of mycophenolic acid in Nagase analbuminemic rats: Evaluation of protein binding effects using the modeling and simulation approach.

This study aimed to examine the pharmacokinetics and pharmacodynamics of mycophenolic acid (MPA) in Nagase analbuminemic rats (NARs) to evaluate the effect of protein binding on the associated inosine-5'-monophosphate dehydrogenase (IMPDH) activity. Free fractions of MPA in the control rats and NARs were 2.09 and 24.8%, respectively. Pharmacokinetic and pharmacodynamic parameters simultaneously obtained by the nonlinear mixed effects modeling program NONMEM explained reasonably well the concentrations of MPA and MPA glucuronide as well as IMPDH activity in both rats. NARs showed a higher clearance and a smaller volume of distribution based on the free MPA concentration than the controls did, besides the increase in free fraction. The half-maximal inhibitory concentration based on free MPA was estimated as 163 ng/mL for both rats. Simulations based on the obtained pharmacokinetic and pharmacodynamic parameters showed that the area under the IMPDH activity-time curve decreased non-linearly according to the increase in free fraction of MPA. In conclusion, the experimental data obtained from NARs followed by the modeling and simulation approach quantitatively clarified that the free MPA concentration was suitable for the biomarker of immunosuppressive effect of MPA. Dose adjustments based on the total MPA may cause unnecessary overexposure to MPA in patients with hypoalbuminemia.

Sensitive and validated LC-MS/MS methods to evaluate mycophenolic acid pharmacokinetics and pharmacodynamics in hematopoietic stem cell transplant patients.

Monitoring of pharmacodynamics in addition to pharmacokinetics is one of strategies to individualize mycophenolate mofetil therapy. The purpose of this study was to develop sensitive liquid chromatography-tandem mass spectrometry (LC-MS/MS) methods for evaluation of the pharmacokinetics and pharmacodynamics of mycophenolic acid (MPA). Concentrations of mycophenolic acid glucuronide (MPAG), mycophenolic acid acyl-glucuronide, as well as unbound MPA and MPAG, were determined, and inosine-5'-monophosphate dehydrogenase activity was calculated by measuring concentrations of produced xanthosine-5'-monophosphate (XMP) and intracellular adenosine-5'-monophosphate after incubation of peripheral blood mononuclear cell (PBMC) lysates. A metal-free Mastro(TM) column and two gradient patterns were used to improve the quantification limit of XMP to 0.1 µM. In the clinical MPA concentration range, the linearity of the calibration curve, inter- and intra-day precision and accuracy satisfied the relevant US Food and Drug Administration guidelines. The MPA concentrations in hematopoietic stem cell transplant (HSCT) patients determined by the enzyme assay and the present LC-MS/MS method showed a good correlation (r(2) = 0.95, p < 0.001). In this study, we report sensitive and validated LC-MS/MS methods to evaluate the pharmacokinetics and pharmacodynamics of MPA, which are sufficiently sensitive to assess small quantities of PBMC lysates collected shortly after HSCT.

Development of asymptomatic acquired factor V inhibitor after the administration of antibiotics.

Acquired factor V (FV) inhibitor is a rare coagulation disorder, the causes and clinical symptoms of which are known to vary widely. Acquired FV inhibitor mostly occurs with exposure to fibrin glues during surgical procedures. We experienced a case with asymptomatic acquired FV inhibitor caused by antibiotic therapy for aspiration pneumonia.A man in his eighties had been treated for chronic atrial fibrillation, cerebral infarction and spinal canal stenosis. He was admitted after antibiotic therapy for aspiration pneumonia in a previous hospital. On admission, the screening test for blood coagulation revealed extreme prolongation of both prothrombin time (PT) and activated partial thromboplastin time (APTT). Vitamin K was administered intravenously after cessation of warfarin, but neither PT nor APTT showed any improvement. Subsequently, a cross mixing test was performed and showed an inhibitor pattern. Furthermore, a high titer of FV inhibitor was detected by the Bethesda method and an acquired FV inhibitor was thus diagnosed. Despite the presence of FV inhibitor, the patient showed no bleeding symptoms. Eight months later, both PT and APTT had diminished to normal clotting time values without immunosuppressive therapies.

Retrospective study of tension-free vaginal mesh operation outcomes for prognosis improvement.

AIM: The aim of this study is to look at the weak points of tension-free vaginal mesh (TVM) operation for pelvic organ prolapse by reviewing recurrent cases. METHODS: Five hundred and twenty-six transobturator TVM operations were performed at the University of Occupational and Environmental Health hospital from August 2006 to December 2011. Thirty-seven patients with pelvic organ prolapse quantification (POP-Q) stage II or higher after their initial operations were diagnosed as recurrent and were enrolled in the present study. The patients' characteristics, procedures of the initial and second operations, and the recurrence rates in the pelvic organs were retrospectively reviewed. RESULTS: The median age, parity and body mass index of the patients were 66.5 years (range, 48-80), 2.2-times (range, 1-5) and 24.1 kg/m(2) (range, 17.6-32.1), respectively. The mean/median follow-up periods of the whole group were 29.7/35.9 months and the recurrent prolapse rate was 7.0% (37/526 cases). We performed reoperations in eight cases (8/526 cases; 1.5%). The median recurrent period after the initial operations was 6.9 months (range, 1-24). All recurrent cases showed severe prolapse with POP-Q stage III to IV before the initial operations. The uterus showed the most frequent recurrent prolapse after anterior posterior TVM or anterior TVM. CONCLUSION: Because the uterus is the most recurrent organ after TVM operations, TVM operations may not be sufficient for apical suspension.

Organic luminescent molecule with energetically equivalent singlet and triplet excited states for organic light-emitting diodes.

We demonstrate an organic molecule with an energy gap between its singlet and triplet excited states of almost zero (ΔE(ST) ∼ 0 eV). Such separation was realized through proper combination of an electron-donating indolocarbazole group and a diphenyltriazine electron-accepting moiety. Calculated and measured ΔE(ST) were 0.003 and 0.02 eV, respectively. A total photoluminescence efficiency of 59% ± 2% with 45% ± 2% from a delayed component and 14% ± 2% from a prompt component was obtained for a doped film. Organic light emitting diodes containing this molecule as an emitting dopant exhibited an unexpectedly high external electroluminescence efficiency of η(EQE) = 14% ± 1%.

Guidelines for office gynecology in Japan: Japan Society of Obstetrics and Gynecology and Japan Association of Obstetricians and Gynecologists 2011 edition.

Gynecology in the office setting is developing worldwide. Clinical guidelines for office gynecology were first published by the Japan Society of Obstetrics and Gynecology and the Japan Association of Obstetricians and Gynecologists in 2011. These guidelines include a total of 72 clinical questions covering four areas (Infectious disease, Malignancies and benign tumors, Endocrinology and infertility, and Healthcare for women). These clinical questions were followed by several answers, backgrounds, explanations and references covering common problems and questions encountered in office gynecology. Each answer with a recommendation level of A, B or C has been prepared based principally on evidence or consensus among Japanese gynecologists.These guidelines would promote a better understanding of the current standard care practices for gynecologic outpatients in Japan.

Intravaginal microbial flora by the 16S rRNA gene sequencing.

OBJECTIVE: Conventional diagnosis of bacterial vaginosis contains some controversial points. To understand accurately the relationship between clinical stages and the microbiotas, the intravaginal microbial flora was analyzed by the clone library method. STUDY DESIGN: Vaginal fluid samples from 31 patients were examined. Lactobacillary grade, Nugent score, culture-based method, and clone library analysis using the 16SrRNA gene sequencing were performed and were compared with each other. RESULTS: Patients were categorized by Lactobacillary grade as I (normal) (n = 6), II (intermediate) (n = 11), and III (bacterial vaginosis) (n = 14). The clone library analysis detected 36 bacterial genera and 60 species from all 31 samples. A principal component analysis of the microbial proportions revealed a novel classification, which suggested the significance of the relative ratio of Lactobacillus iners, Atopobium vaginae and anaerobes in bacterial vaginosis. CONCLUSION: Clone library analysis in combination with the conventional method provides substantial information for diagnosis of bacterial vaginosis.