BACKGROUND: The multiplex real-time PCR assay is a sensitive test for simultaneous detection of various pathogens of sexually transmitted infections (STIs). We evaluated the performance of two multiplex real-time PCR assays for six STI pathogens. METHODS: DNA samples after being used to conduct PCR for STI pathogens were stored below −70℃. Chlamydia trachomatis (CT), Neisseria gonorrhoeae (NG), Mycoplasma genitalium (MG), Mycoplasma hominis (MH), Ureaplasma urealyticum (UU), and Trichomonas vaginalis (TV) were detected by multiplex real-time PCR with GeneFinder STD I (CT/NG/UU)/II (MG/MH/TV) Multiplex Real-time PCR Kits (Infopia, Korea; GeneFinder assay) and Real-Q CT&NG/MH&TV/MG&UU Kits (BioSewoom, Korea; Real-Q assay). Discrepant results were resolved by another multiplex real-time assay, Anyplex II STI-7 Detection (Seegene, Korea). Any two positive results for the assays were considered true positive. RESULTS: Among 81 samples, the GeneFinder assay detected 63 pathogens from 45 cases (16 CT, 2 NG, 6 MG, 20 MH, 18 UU, and 1 TV) and Real-Q assay detected 66 pathogens from 47 cases (16 CT, 2 NG, 8 MG, 20 MH, 19 UU, and 1 TV). For the results of positive cases and negative cases, the overall concordance rate between the two multiplex real-time assays was 93.8% (Kappa=0.87). For each pathogen, the agreement rates of the two assays ranged from 97.5 to 100% (Kappa>0.8). CONCLUSION: There was no significant difference between the results of GeneFinder assay and Real-Q assay. Both multiplex real-time PCR assays can be useful methods for the detection of STI pathogens in clinical laboratories.
Chlamydia trachomatis ; DNA ; Korea ; Mycoplasma genitalium ; Mycoplasma hominis ; Neisseria gonorrhoeae ; Polymerase Chain Reaction ; Real-Time Polymerase Chain Reaction* ; Sexually Transmitted Diseases* ; Trichomonas vaginalis ; Ureaplasma urealyticum

There is limited research on sexually transmitted infections (STIs) among adolescents in Korea. The objective of this study was to explore the prevalence of and risk factors for STIs among Korean adolescents under probation. A cross-sectional analysis was conducted in one juvenile-delinquent center and five probation offices in Korea to determine the prevalence of STIs caused by the following pathogens: Chlamydia trachomatis, Neisseria gonorrhoeae, Trichomonas vaginalis, herpes simplex virus (HSV), human immunodeficiency virus (HIV), Treponema pallidum, Mycoplasma hominis, Mycoplasma genitalium, Ureaplasma urealyticum, and Ureaplasma parvum. Of the 237 (208 male and 29 female) participating adolescents, 152 (64.1%) had a history of coitus. Overall, 133 (56.1%) subjects tested positive for at least one microorganism in their genitourinary tract. The most prevalent pathogen was U. urealyticum (24.7%, n = 65), followed by U. parvum (24.1%, n = 57), M. hominis (17.3%, n = 41), C. trachomatis (13.9%, n = 33), N. gonorrhoeae (1.7%, n = 4), T. vaginalis (0.8%, n = 2), and HSV (0.8%, n = 2). The prevalence of syphilis was 0.8% (n = 2). There were no reported cases of HIV infection. Fifty-four participants (35.5%) were positive with more than two pathogens. We did not find any significant difference between STIs and socioeconomic factors, behavioral factors or sexual practices. In conclusion, the prevalence of STIs among adolescents under probation was high. Systematic screening programs, more practical sexual education, and adequate provision of treatment are essential for the prevention and management of STIs among adolescents, especially those under probation.
Adolescent* ; Chlamydia trachomatis ; Coitus ; Cross-Sectional Studies ; Education ; HIV ; HIV Infections ; Humans ; Korea ; Male ; Mass Screening ; Mycoplasma ; Mycoplasma genitalium ; Mycoplasma hominis ; Neisseria gonorrhoeae ; Prevalence* ; Risk Factors* ; Sexually Transmitted Diseases* ; Simplexvirus ; Socioeconomic Factors ; Syphilis ; Treponema pallidum ; Trichomonas vaginalis ; Ureaplasma ; Ureaplasma urealyticum

BACKGROUND: Many molecular diagnostic methods have been developed to detect sexually transmitted infections (STI). The STDetect Chip (LabGenomics, Korea) which is a DNA microarray-based tool, newly developed for STI diagnosis in vitro, and the real-time PCR-based Anyplex STI-7 (Seegene, Korea) in clinical use were evaluated using ATCC DNA and clinical samples to determine the clinical usefulness of the STDetect Chip. METHODS: The two methods were compared for consistency, sensitivity, and specificity for 6 pathogens in 300 prospectively selected clinical samples. Analytical sensitivity for ATCC Neisseria gonorrhoeae, Chlamydia trachomatis, Mycoplasma hominis and Trichomonas vaginalis DNA and the effect of mixing bacterial DNA were studied. RESULTS: The consistency of the two methods for clinical samples was superior at more than 0.92 kappa value. The sensitivity and specificity of the STDetect Chip compared with Anyplex STI-7 were 90.5-98.8%, and 95.6-99.6%, respectively. With similar analytical performance for ATCC DNA, the STDetect Chip detected 10(-5) ng/µL of N. gonorrhoeae, 10(-4) ng/µL of C. trachomatis, 10(-6) ng/µL of M. hominis, and 10(-3) ng/µL of T. vaginalis. For the mixture of three bacterial DNAs, less sensitive detection level was observed for T. vaginalis. CONCLUSIONS: The STDetect Chip showed good agreement with the Anyplex STI-7 test and it is considered clinically useful for detecting sexually transmitted pathogens.
Chlamydia trachomatis ; Diagnosis* ; DNA* ; DNA, Bacterial ; In Vitro Techniques ; Mycoplasma hominis ; Neisseria gonorrhoeae ; Oligonucleotide Array Sequence Analysis ; Pathology, Molecular ; Polymerase Chain Reaction ; Prospective Studies ; Sensitivity and Specificity ; Sexually Transmitted Diseases* ; Trichomonas vaginalis

BACKGROUND: Sexually transmitted infections (STI) encompass a variety of clinical syndromes caused by many pathogens that are transmitted through sexual activity. Multiplex PCR is frequently used to detect STI. In this study, two multiplex real-time PCR-based assays were used to detect STI in clinical specimens, and the concordance of the results obtained by each method was evaluated. METHODS: A total of 626 specimens were tested using the Anyplex II STI-7 (Seegene, Korea) and Seeplex STD6 ACE Detection kits (Seegene). RESULTS: Among the 626 individuals tested, 227 (44.2%) tested positive for STI by using Anyplex II STI-7. The prevalence rates of the various infectious microorganisms detected were as follows: Chlamydia trachomatis (C. trachomatis), 19.2% (120/626); Neisseria gonorrhoeae (N. gonorrhoeae), 5.6% (35/626); Trichomonas vaginalis (T. vaginalis), 0.2% (1/626); Mycoplasma genitalium (M. genitalium), 8.1% (51/626); Mycoplasma hominis (M. hominis), 2.9% (18/626); Ureaplasma urealyticum (U. urealyticum), 17.6% (110/626); and Ureaplasma parvum, 3.7% (23/626). The concordance rates for the STI-7 and STD6 assays in detecting the various types of microorganism were as follows: C. trachomatis, (99.5%); N. gonorrhoeae, (99.7%); T. vaginalis, (100%); M. genitalium, (100%); M. hominis, (100%); and U. urealyticum (99.2%). CONCLUSIONS: A high degree of concordance was observed between the results obtained using the Anyplex II STI-7 kits and those obtained using the Seeplex STD6 ACE Detection kits.
Chlamydia trachomatis ; Methods ; Multiplex Polymerase Chain Reaction ; Mycoplasma genitalium ; Mycoplasma hominis ; Neisseria gonorrhoeae ; Prevalence ; Sexual Behavior ; Sexually Transmitted Diseases ; Trichomonas vaginalis ; Ureaplasma ; Ureaplasma urealyticum

BACKGROUND: Pelvic inflammatory disease (PID) is a microbial infection caused by the upward spread of infectious organisms through the cervical os. Early diagnosis and treatment of PID are essential for the prevention of sequelae such as ectopic pregnancies, infertility, and chronic pelvic pain. Although Chlamydia trachomatis and Neisseria gonorrhoeae are well-known causal agents of PID, there have been reports on some changes in PID-associated infection. The aim of this study was to investigate the infection patterns in patients with PID in Jeju. METHODS: Endocervical samples obtained from 65 patients with PID were tested for C. trachomatis, Mycoplasma genitalium, Mycoplasma hominis, N. gonorrhoeae, Trichomonas vaginalis, and Ureaplasma urealyticum using multiplex PCR. RESULTS: The samples were positive for C. trachomatis (63%), M. hominis (34%), U. urealyticum (20%), M. genitalium (17%), N. gonorrhoeae (9%), and T. vaginalis (6%). CONCLUSIONS: This study showed that C. trachomatis infection was prevalent and the incidence of M. hominis was higher than that of U. urealyticum.
Chlamydia trachomatis ; Early Diagnosis ; Female ; Humans ; Incidence ; Infertility ; Mycoplasma genitalium ; Mycoplasma hominis ; Neisseria gonorrhoeae ; Pelvic Inflammatory Disease ; Pelvic Pain ; Polymerase Chain Reaction ; Pregnancy ; Pregnancy, Ectopic ; Trichomonas vaginalis ; Ureaplasma urealyticum

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

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

PURPOSE: Our purpose was to conduct a screening test for urethritis or cervicitis as a sexually transmitted disease (STD) by using multiplex polymerase chain reaction(PCR) and to determine the prevalence of Chlamydia trachomatis, Neisseria gonorrhoeae, Mycoplasma genitalium, Ureaplasma urealyticum, Mycoplasma hominis, and Trichomonas vaginalis in asymptomatic people. MATERIALS AND METHODS: From July 2010 to December 2010, 709 persons who came to the hospital for a general checkup were tested. Multiplex PCR assays were done with first voided urine samples or endocervical swabs by use of the Seeplex(R) STD6 ACE Detection kit. RESULTS: The mean age in this study was 45.4+/-8.1 years. Among the 709 persons, 229 (32.3%) had a positive result for at least one microorganism, 48 (6.8%) had two different species, 6 (0.8%) had three different species, and 1 person had four different species. The overall prevalence of asymptomatic STDs such as urethritis or cervicitis was 7.1% (50/709). The prevalence rates of chlamydia trachomatis, Neisseria gonorrhoeae, Mycoplasma genitalium, Ureaplasma urealyticum, Mycoplasma hominis, and Trichomonas vaginalis infection in asymptomatic persons were 5.6% (40/709), 0.4% (3/709), 0.3% (2/709), 22.1% (157/709), 11.6% (82/709), and 1.1% (8/709), respectively. CONCLUSIONS: With only a single sample, we could identify the prevalence rates of six microorganisms and the overall proportion of urethritis or cervicitis in asymptomatic people. This proportion cannot be neglected; therefore, screening tests for sexually transmitted diseases such as urethritis or ervicitis should be recommended to asymptomatic people.
Chlamydia ; Chlamydia trachomatis ; Humans ; Mass Screening ; Multiplex Polymerase Chain Reaction ; Mycoplasma ; Mycoplasma genitalium ; Mycoplasma hominis ; Neisseria gonorrhoeae ; Polymerase Chain Reaction ; Porphyrins ; Prevalence ; Sexually Transmitted Diseases ; Trichomonas vaginalis ; Ureaplasma ; Ureaplasma urealyticum ; Urethritis ; Uterine Cervicitis

We isolated 30 Trichomonas vaginalis for the PCR detection from the gynecological outpatients in the Affiliated Hospital of Zhengzhou University using the specific 16s rDNA primers of Mycoplasma hominis. The results showed that there were 25 cases of Mycoplasma hominis infection, with the infection rate of 83.33%. This gave a clew that the symbiosis of Trichomonas vaginalis with Mycoplasma hominis may be of certain generality in China. We sequenced the ferredoxin gene of 10 Trichomonas vaginalis where 5 Mycoplasma hominis were positive and five negative, and found that the ferredoxin (Fd) gene of the 10 Trichomonas vaginalis were exactly the same. But compared to the genes in the GenBank, a comparative analysis of the gene revealed that there were 3 more ctg bases at the 200th position of encoding leucine, but this did not lead to changes in reading frame. The gene homology was 99%.
Amino Acid Sequence ; Base Sequence ; Female ; Ferredoxins ; genetics ; Humans ; Molecular Sequence Data ; Mycoplasma hominis ; genetics ; physiology ; Symbiosis ; genetics ; Trichomonas vaginalis ; genetics ; physiology

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