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: Real-time polymerase chain reaction (PCR) is generally regarded as a very accurate and time-saving method, but it is expensive to run. We evaluated the reliability of an inexpensive and a researcher-friendly gel electrophoresis-based PCR method for the quantification of mRNA, and the results were compared with those obtained by real-time PCR. METHODS: We compared the results of relative quantification for MMP-1 measured by real-time PCR and by ethidium bromide stained-agarose gel electrophoresis after end-point PCR. RESULTS: There was significant but very weak correlation between real-time PCR and end-point PCR for relative quantification of MMP-1 (r=0.16, P<0.01). CONCLUSIONS: Our results suggest that the use of the gel electrophoresis-based end-point PCR is inappropriate for quantifying mRNA. Therefore, in order to confirm the result of relative quantification by end-point PCR, the newly established real-time PCR method or northern hybridization should be applied.
DNA* ; Electrophoresis ; Electrophoresis, Agar Gel* ; Ethidium ; Polymerase Chain Reaction ; Real-Time Polymerase Chain Reaction* ; RNA, Messenger ; Sepharose*

OBJECTIVE: A recent study suggested that a single nucleotide polymorphism (SNP) at position nt 9250 (C to T) in exon 7 of the osteopontin (OPN) gene is strongly associated with the susceptibility to systemic lupus erythematosus (SLE). This study examined the possible association between a single nucleotide polymorphism (SNP) at position nt 9250 (C to T) and SLE and measured the serum levels of OPN in Korean patients with SLE. METHODS: A total of 39 patients with SLE and 104 healthy controls were enrolled in this study. SNP located at position 9250 in the OPN gene were genotyped using the restriction fragment length polymorphism (RFLP). The serum levels of OPN in 39 patients with SLE and 20 healthy controls were determined by enzyme-linked immunosorbent assay. RESULTS: The allele frequencies of C and T at this position in patients with SLE were 34.6 and 65.4, whereas those in the controls were 20.7 and 79.3 (p<0.05). The serum levels of OPN in 39 patients with SLE were significantly higher than that in 20 healthy controls (49.13+/-26.71 versus 28.49+/-18.39 ng/ml, p<0.05). The increase in OPN concentration was associated with the SLE disease activity index (SLEDAI) score in all SLE patients (r=0.337, p<0.05). CONCLUSION: The allele frequencies of Eta-1/osteopontin were significantly associated with SLE. Moreover, the increased serum level of OPN is associated with the SLE disease activity. However, further investigation in larger groups in Korea will be needed.
Enzyme-Linked Immunosorbent Assay ; Exons ; Gene Frequency ; Humans ; Korea ; Lupus Erythematosus, Systemic ; Osteopontin ; Polymorphism, Genetic ; Polymorphism, Restriction Fragment Length ; Polymorphism, Single Nucleotide

Candidemia due to uncommon Candida spp. appears to be increasing in incidence. C. dubliniensis has been increasingly recovered from individuals not infected with HIV. Identification of C. dubliniensis can be problematic in routine clinical practice due to its phenotypic resemblance to C. albicans. We report the first case of C. dubliniensis candidemia in Korea, which occurred in a 64-yr-old woman who presented with partial seizure, drowsiness, and recurrent fever. Germ-tube positive yeast that was isolated from blood and central venous catheter tip cultures formed smooth, white colonies on sheep blood agar and Sabouraud agar plates, indicative of Candida spp. C. dubliniensis was identified using the Vitek 2 system (bioMerieux, USA), latex agglutination, chromogenic agar, and multiplex PCR. The blood isolate was susceptible to flucytosine, fluconazole, voriconazole, and amphotericin B. After removal of the central venous catheter and initiation of fluconazole treatment, the patient's condition gradually improved, and she was cleared for discharge from our hospital. Both clinicians and microbiologists should be aware of predisposing factors to C. dubliniensis candidemia in order to promote early diagnosis and appropriate treatment.
Amphotericin B/pharmacology ; Antifungal Agents/pharmacology/therapeutic use ; Candida/drug effects/*isolation & purification ; Candidemia/*diagnosis/drug therapy ; Catheterization, Central Venous ; Female ; Fluconazole/pharmacology/therapeutic use ; Flucytosine/pharmacology ; Humans ; Microbial Sensitivity Tests ; Middle Aged ; Pyrimidines/pharmacology ; Triazoles/pharmacology

BACKGROUND: Recently, the disk diffusion testing of fluconazole against Candida spp. has been attempted in order to provide a simple inexpensive method in the routine laboratory. We investigated the possibility and reliability of a fluconazole disk diffusion method using a Mueller-Hinton agar supplemented with glucose and methylene blue (GM-MH). METHODS: One hundred and seven isolates of Candida spp. (54 C. albicans, 21 C. glabrata, 20 C. tropicalis, 6 C. parapsilosis, 4 C. krusei, and 2 C. lusitaniae) were tested with the broth microdilution method of National Committee for Clinical Laboratory Standards (NCCLS) document M27-A2 and a disk diffusion test using GM-MH agar. RESULTS: The overall categorical agreement between the NCCLS method and the disk diffusion method was 89.8% for fluconazole, with 0.9% very major errors and 9.3% minor errors; no major errors were detected. CONCLUSIONS: These data suggest that the fluconazole disk diffusion test on GM-MH agar can be used as a routine screening procedure for susceptibility of Candida spp. in the clinical laboratory.
Agar* ; Candida* ; Diffusion* ; Fluconazole* ; Glucose ; Mass Screening ; Methylene Blue

BACKGROUND: Polymerase chain reacation (PCR)-based methods have been described for rapid detection and identification of Candida spp. Multiplex PCR assay was developed using internal transcribed spacers and topoisomerase II gene for the accurate identification of Candida species. METHODS: We designed Dual Specificity Oligo (DSO) primers for multiplex PCR. Multiplex PCR was followed by agarose gel electrophoresis to test 8 type strains (C. albicans, C. parapsilosis, C. glabrata, C. tropicalis, C. krusei, C. guilliermondii, C. lusitaniae, C. dubliniensis) and 96 clinical isolates (C. albicans 51 isolates, C. parapsilosis 10 isolates, C. glabrata 10 isolates, C. tropicalis 9 isolates, C. krusei 6 isolates, C. guilliermondii 5 isolates, C. lusitaniae 5 isolates) of Candida spp. RESULTS: With multiplex PCR using DSO primers, the eight Candida type strains each could be easily differentiated and all 96 clinical isolates were identified as the same species as were identified by the conventional method. CONCLUSION: Multiplex PCR followed by electrophoresis can be useful for the simple and rapid identification of Candida species in routine laboratories.
Candida* ; DNA Topoisomerases, Type II ; Electrophoresis ; Electrophoresis, Agar Gel ; Multiplex Polymerase Chain Reaction* ; Sensitivity and Specificity

BACKGROUND: The genus Candida comprises 163 species, the most common pathogen in the genus is C. albicans, however, other Candida species are considered as emergent pathogens. Because there are species-specific differences in the susceptibility of Candida spp. to the currently used therapeutic drugs, species identification is critical for therapeutic planning and accurate epidemiological records. The objective of this study is to evaluate the performance of tRNA intergenic length polymorphism (tDNA-ILP) analysis for the accurate identification of Candida species. METHODS: For the identifying the 7 type strains and 54 clinical isolates of Candida, the suitability of tDNA-ILP was evaluated. The polymerase chain reaction (PCR) using a pair of primers or a single primer was performed. The PCR products were separated by electrophoresis in 2% agarose gels for 70 min. RESULTS: In seven Candida type strains, tDNA-ILP using a single primer (reverse) can be easily analyzed by visual comparision. Fifty-three of 54 strains were identified as the same species with conventional identification. CONCLUSIONS: The tDNA-ILP analysis can be useful for the simple and rapid identification of Candida species in routine laboratories.
Candida* ; Electrophoresis ; Gels ; Polymerase Chain Reaction ; RNA, Transfer ; Sepharose

No abstract available.
Dasatinib ; Leukemia, Myelogenous, Chronic, BCR-ABL Positive ; Primary Myelofibrosis

PURPOSE: Chronic prostatitis frequently occurs in men of all ages. Recent studies suggest that fastidious microorganisms may play a role in chronic prostatitis/chronic pelvic pain syndrome (CP/CPPS). The aim of this study was to evaluate the usefulness and significance of multiplex polymerase chain reaction (PCR) in the diagnosis of CP/CPPS. MATERIALS AND METHODS: First voided urine (FVU) and/or expressed prostatic secretions (EPS) were collected from 92 patients. Multiplex PCR, using Dual Specificity Oligo (DSO(TM)) primers, was used to test for Chlamydia trachomatis (CT), Mycoplasma genitalium (MG), Mycoplasma hominis (MH), Neisseria gonorrhoeae (NG), Trichomonas vaginalis (TV) and Ureaplasma urealyticum (UU). RESULTS: Multiplex PCR can be easily analyzed via visual comparison. Nine (39.1%) of the 23 CP/CPPS IIIa and 12 (17.4%) of the 69 IIIb patients had positive multiplex PCR, with a total of 27 microorganisms isolated, including CT, MH, MG, UU, TV and NG in 9, 7, 4, 4, 2 and 1 case, respectively. Co-infections with 2 or 3 organisms occurred in 5 cases. For the samples collected from 32 patients for both FVU and EPS, 68.7% gave the same results. CONCLUSIONS: Multiplex PCR, using DSO(TM) primers, can be useful for the simple detection of fastidious microorganisms in CP/CPPS. To achieve reliable results with multiplex PCR, feasible guidelines and standardization are of major importance. Further studies will be required to define the usefulness of molecular tests for CP/CPPS in clinical practice.
Chlamydia trachomatis ; Coinfection ; Diagnosis ; Humans ; Male ; Multiplex Polymerase Chain Reaction* ; Mycoplasma genitalium ; Mycoplasma hominis ; Neisseria gonorrhoeae ; Pelvic Pain ; Polymerase Chain Reaction ; Prostatitis* ; Sensitivity and Specificity ; Trichomonas vaginalis ; Ureaplasma urealyticum

Tuberculosis (Tb) is a common disease in the developing world and its incidence is slowly increasing in developed countries, where a resurgence has occurred subsequent to the acquired immunodeficiency syndrome epidemic. In addition, patients with end-stage renal disease who are on maintenance hemodialysis carry a high risk for Tb; reported incidence varies from 6-16 times that of the general population. Extrapulmonary Tb constitutes a major part of Tb in dialysis patients. Isolated pancreatic Tb is a very rare occurrence in the setting of extrapulmonary Tb. It usually occurs as a complication of miliary Tb in immunodeficient individuals, particularly those with human immunodeficiency virus infection. There is no isolated pancreatic Tb in patients with end-stage renal disease. We recently experienced a case of isolated pancreatic Tb diagnosed by acid fast bacilli culture, Tb polymerase chain reaction from ultrasound-guided fine needle aspiration, and an excellent response after anti-Tb treatment in a 72-year-old patient with end-stage renal disease.
Acquired Immunodeficiency Syndrome ; Aged ; Biopsy ; Biopsy, Fine-Needle ; Developed Countries ; Dialysis ; HIV ; Humans ; Incidence ; Kidney Failure, Chronic ; Pancreas ; Polymerase Chain Reaction ; Renal Dialysis ; Tuberculosis