OBJECTIVETo investigate the distribution of pathogenic C.albican genotype and Candida species in association with the severity of vulvovaginal candidiasis (VVC).

METHODSPolymerase chain reaction-single strand conformation polymorphism (PCR-SSCP) of the internal transcribed spacer analysis was employed to identify the Candida species isolated from the vaginal secretions of 198 patients with acute VVC. SSCP and GeneScan analyses of microsatellite locus I polymorphism were used to determine the genotypes of the clinical isolates of C. albican associated with VVC. All the patients were scored for clinical signs and symptoms to evaluate the severity of VVC.

RESULTSA total of 198 Candida strains were isolated from VVC patients, including 140 (70.7%) C. albicans strains and 58 (29.3%) non-albicans strains. In the 95 patients with severe VVC and 103 with mild-moderate VVC, C.albican was detected in 62.1% and 76.6% of the patients, respectively (P=0.011). Thirty-eight microsatellite locus I genotypes were detected in 140 unrelated C. albican strains, among which the dominant genotypes 30-45 (44 strians, 31.43%) and 32-46 (23 strains, 16.43%) were the most common, followed by genotypes 30-46 (4 strains, 2.86%) and 32-47 (9 strains, 6.42%). The overall frequencies of the 4 genotypes were significantly higher in severe VVC than in mild-moderate VVC cases (77.9% vs 42.0%, P<0.001).

CONCLUSIONC. albicans remains the most common pathogenic Candia species in patients with VVC, but the non-alibcans species seem more likely to cause severe VVC. The dominant genotypes of C. albicans with a tropism for the vagina are correlated to the severity of VVC.

Adolescent ; Adult ; Candida ; classification ; isolation & purification ; Candida albicans ; genetics ; isolation & purification ; Candidiasis, Vulvovaginal ; microbiology ; Female ; Genotype ; Humans ; Middle Aged ; Polymerase Chain Reaction ; methods ; Polymorphism, Single-Stranded Conformational ; Severity of Illness Index ; Young Adult

To find a fast and efficient way of identifying seven common dermatophytes in clinical practice, we used the techniques of polymerase chain reaction (PCR) and PCR-restriction fragment length polymorphism (RFLP) targeting Topoisomerase II gene. The DNA of 7 dermatophytes, along with Candida albicans, Aspergillus terreus and Aspergillus flavus were amplified by consensus primer dPsD1. They were then subjected to a second PCR with primers dPsD2 and species-specific primers PsT and PsME separately. 6 of the products generated by dPsD2 were digested with restriction enzyme Hinc II. DNA fragments of 3390 bp and 2380 bp was amplified by using consensus primer dPsD1 and dPsD2 from the genomic DNA of each dermatophyte species separately. By combining the results of the two species-specific primer sets (PsT and PsME), all species of dermatophyte yielded unique sizes-set of PCR products expect for T. mentagrophytes and T. tonsurans. From the restriction profiles of Hinc II, 6 of the 7 dermatophytoses were diagnosed to species level including T. mentagrophytes and T. tonsurans. By combining the results of the PCR and PCR-RFLP, the 7 common dermatophytes can be identified to species level. It is conclude that the multiplex PCR and PCR-RFLP identification targeting the DNA topoisomerase II gene is rapid and efficient.
Arthrodermataceae ; classification ; isolation & purification ; Aspergillus ; isolation & purification ; Candida albicans ; isolation & purification ; DNA Topoisomerases, Type II ; genetics ; Dermatomycoses ; microbiology ; Humans ; Polymerase Chain Reaction ; Polymorphism, Restriction Fragment Length ; Trichophyton ; isolation & purification

To find a fast and efficient way of identifying seven common dermatophytes in clinical practice, we used the techniques of polymerase chain reaction (PCR) and PCR-restriction fragment length polymorphism (RFLP) targeting Topoisomerase II gene. The DNA of 7 dermatophytes, along with Candida albicans, Aspergillus terreus and Aspergillus flavus were amplified by consensus primer dPsD1. They were then subjected to a second PCR with primers dPsD2 and species-specific primers PsT and PsME separately. 6 of the products generated by dPsD2 were digested with restriction enzyme Hinc II. DNA fragments of 3390 bp and 2380 bp was amplified by using consensus primer dPsD1 and dPsD2 from the genomic DNA of each dermatophyte species separately. By combining the results of the two species-specific primer sets (PsT and PsME), all species of dermatophyte yielded unique sizes-set of PCR products expect for T. mentagrophytes and T. tonsurans. From the restriction profiles of Hinc II, 6 of the 7 dermatophytoses were diagnosed to species level including T. mentagrophytes and T. tonsurans. By combining the results of the PCR and PCR-RFLP, the 7 common dermatophytes can be identified to species level. It is conclude that the multiplex PCR and PCR-RFLP identification targeting the DNA topoisomerase II gene is rapid and efficient.
Arthrodermataceae/classification ; Arthrodermataceae/*isolation & purification ; Aspergillus/*isolation & purification ; Candida albicans/isolation & purification ; DNA Topoisomerases, Type II/genetics ; Dermatomycoses/microbiology ; Polymerase Chain Reaction ; Polymorphism, Restriction Fragment Length ; Trichophyton/*isolation & purification

BACKGROUND: We evaluated the efficacy of multilocus sequence typing (MLST) for assessing the genetic relationship among Candida albicans isolates from patients with candidemia in a hospital setting. METHODS: A total of 45 C. albicans isolates from 21 patients with candidemia were analyzed. The MLST results were compared with results obtained by Southern blot hybridization (C1 fingerprinting) and pulsed-field gel electrophoresis (PFGE). PFGE analysis included karyotyping and restriction endonuclease analysis of genomic DNAs using BssHII (REAG-B) and SfiI (REAG-S). RESULTS: The 45 isolates yielded 20 unique diploid sequence types (DSTs) by MLST, as well as 12 karyotypes, 15 REAG-B patterns, 13 REAG-S patterns, and 14 C1 fingerprinting types. Microevolution among intra-individual isolates was detected in 6, 5, 3, 5, and 7 sets of isolates by MLST (1 or 2 allelic differences), REAG-B, REAG-S, C1 fingerprinting, and a combination of all methods, respectively. Among 20 DSTs, 17 were unique, and 3 were found in more than 1 patient. The results of 2 DSTs obtained from 9 patient isolates were in agreement with REAG and C1 fingerprinting patterns. However, the remaining DST, which was shared by 2 patient isolates, showed 2 different PFGE and C1 fingerprinting patterns. In addition, 3 sets of isolates from different patients, which differed in only 1 or 2 alleles by MLST, also exhibited different PFGE or C1 fingerprinting patterns. CONCLUSIONS: MLST is highly discriminating among C. albicans isolates, but it may have some limitations in typing isolates from different patients, which may necessitate additional analysis using other techniques.
Alleles ; Blotting, Southern ; Candida albicans/*classification/genetics/isolation & purification ; Candidemia/*microbiology ; DNA, Fungal/*analysis ; Electrophoresis, Gel, Pulsed-Field ; Genotype ; Humans ; Karyotyping ; Multilocus Sequence Typing/*methods

OBJECTIVETo investigate the candidal carriage and the Candida species in HIV positive patients and to explore the relationship between oral candidal carriage and oral candidiasis.

METHODSSixty-four HIV positive patients and 42 healthy controls were included in this study. Oral rinse technique was used to detect the candidal carriage. The isolates were identified using multiple measures, including Gram staining reaction, chlamydospore, pseudo-hyphal and hyphal production test, CHROMagar Candida test and API 20 C AUX yeast identification system.

RESULTSThirty-nine of 64 HIV positive cases were diagnosed as oral candidiasis. Seventy-four Candida strains were isolated from 52 of 64 HIV positive cases, only 7 strains were isolated from 42 healthy controls (P < 0.001). Of the 74 Candida strains isolated from HIV positive cases, 39 were Candida albicans, 15 Candida tropicalis, and 20 other 6 species.

CONCLUSIONSA high prevalence of oral candidiasis and high candidal carriage were found in HIV positive patients compared with those in controls. Candida albicans and Candida tropicalis were the major species. The biotyping of the species isolated from HIV positive patients showed more diversified compared to healthy people, which may suggest the decreased immune ability of the HIV positive patients.

Adult ; Candida ; classification ; isolation & purification ; Candidiasis, Oral ; complications ; diagnosis ; Case-Control Studies ; Female ; HIV Infections ; microbiology ; Humans ; Male ; Middle Aged ; Mouth ; microbiology ; Young Adult

The oral carriage rate of Candida in healthy humans ranges from 40% to 60%. However for a prolonged period, the oral candidal prevalence in humans was documented essentially using data from studies in the West as their prevalence in inhabitants in different regions of the world, including Asia was not known. Yet, recent reports from a number of studies indicate the quality, quantity and prevalence of oral yeasts differ between Asia and other regions for reason that are still unclear. This mini review on such data from Asian studies on oral carriage of Candida provides another intriguing facet of the behavior of this ubiquitous yeast.
Areca ; Asian Continental Ancestry Group ; Candida ; classification ; isolation & purification ; Cohort Studies ; Colony Count, Microbial ; HIV Infections ; microbiology ; Humans ; Leprosy ; microbiology ; Mouth ; microbiology

AIMTo identify heterogeneity of Candida albicans (C. albicans) isolated from the population with cancer in China by using identification medium, subculture molecular typing, and antifungal susceptibility test.

METHODOLOGYOral cheek mucosal specimens from 52 cancer patients receiving chemotherapy were cultured on CHROMagar Candida plates for Candida identification. All the C. albicans colonies on the plates were subcultured and reconfirmed by API20C, then submitted to the antifungal drug susceptibility test with fluconazole and molecular typing using randomly amplified polymorphic DNA-PCR (RAPD) with primers RSD6 and RSD12.

RESULTS54% (28/52) patients were oral yeast carriage in which C. albicans predominated. More than 7 C. albicans colonies were isolated from each of 12 patients (Group A), while less than 5 colonies were isolated from each of 16 patients (Group B). RSD6 and RSD12 were successful in eliciting 17 (A1-A17) and 2 (B1-B2) genotypes, respectively from among the 205 isolates. The two primers were combined to generate 21 genotypes. The C. albicans isolates obtained from the same patient and episode showed a diversity for fluconazole revealed by MIC50 and MIC90.

CONCLUSIONThe heterogeneity of the C. albicans colonies isolated from the same patients can be detected. C. albicans with varied fluconazole susceptibility and genotypic characteristics may coexist in the same oral Candida population.

Adult ; Aged ; Antifungal Agents ; pharmacology ; Candida albicans ; classification ; genetics ; isolation & purification ; Candida glabrata ; classification ; isolation & purification ; Candidiasis, Oral ; microbiology ; China ; DNA, Fungal ; analysis ; Drug Resistance, Fungal ; genetics ; Female ; Fluconazole ; pharmacology ; Genetic Heterogeneity ; Genotype ; Hematologic Neoplasms ; drug therapy ; Humans ; Male ; Microbial Sensitivity Tests ; Middle Aged ; Mouth Mucosa ; microbiology ; Mycology ; methods ; Neoplasms ; drug therapy ; Young Adult

OBJECTIVETo screen antifungal activity of endophytes from Pseudolarix kaempferi.

METHODEndophytes from P. kaempferi were separated. By means of microdilution method, antifungal active endophytes were fast screened by Pyricularia oryzae P-2b model, and activity of endophytes against pathogenic fungus was studied.

RESULT44.8% of endophytes showed activity against P. oryzae P-2b in Pseudolarix kaempferi. Among them JJ314, JJ323 introduced formation of characteristic beads and swellings on the growing hyphae, JJ324 inhibited the conidia germination. They all showed activity against Trichophyton rubrum, Cryptococcus neoformans and Candida albicans.

CONCLUSIONEndophytes from P. kaempferi are a potential resource for the development of antifungal agent.

Antifungal Agents ; isolation & purification ; pharmacology ; Candida albicans ; drug effects ; Cryptococcus ; drug effects ; Fungi ; chemistry ; classification ; isolation & purification ; Mitosporic Fungi ; drug effects ; Pinaceae ; microbiology ; Plants, Medicinal ; microbiology ; Symbiosis ; Trichophyton ; drug effects

Candida haemulonii, one of the non-albicans Candida species, is an emerging yeast pathogen that is known to be resistant to amphotericin B and other antifungal agents such as azoles. These anti-fungal agents have often been associated with clinical treatment failure, so no treatment regimen has been clearly established for invasive C. haemulonii infections. We investigated a catheter-related infection of C. haemulonii candidemia in an adult patient in long-term hospital care. In the early stages, the candidemia remained persistent despite treatment with fluconazole. However, after changing the antifungal agent to caspofungin, the candidemia was resolved. Fluconazole and amphotericin B are not reliable empirical antifungal agents for invasive C. haemulonii infections, as shown in previous case reports. An echinocandin such as caspofungin may be an appropriate empirical choice of antifungal agent for an invasive C. haemulonii infection.
Aged ; Amphotericin B/therapeutic use ; Antifungal Agents/therapeutic use ; Candida/classification/isolation & purification/*pathogenicity ; Candidiasis/drug therapy/*microbiology ; Catheter-Related Infections/drug therapy/*microbiology ; Echinocandins/therapeutic use ; Fluconazole/therapeutic use ; *Hospitals ; Humans ; *Long-Term Care ; Male ; Phylogeny

PURPOSE: The incidence of Candida bloodstream infections (BSI) has increased over the past two decades. The rank order of occurrence and the susceptibility to antifungals of the various Candida species causing BSI are important factors driving the establishment of empirical treatment protocols; however, very limited multi-institutional data are available on Candida bloodstream isolates in Korea. MATERIALS AND METHODS: We investigated the susceptibility to azole antifungals and species distribution of 143 Candida bloodstream isolates recovered from eight university hospitals over a six-month period. Minimal inhibitory concentrations (MICs) of fluconazole, itraconazole, and voriconazole for each isolate were determined by the broth microdilution method of the Clinical and Laboratory Standards Institute (CLSI). RESULTS: The Candida species recovered most frequently from the blood cultures was C. albicans (49%), followed by C. parapsilosis (22%), C. tropicalis (14%), and C. glabrata (11%). The MIC ranges for the Candida isolates were 0.125 to 64microgram/mL for fluconazole, 0.03 to 2microgram/mL for itraconazole, and 0.03 to 1microgram/mL for voriconazole. Overall, resistance to fluconazole was found in only 2% of the Candida isolates (3/143), while the dose-dependent susceptibility was found in 6% (8/143). The resistance and dose-dependent susceptibility of itraconazole were found in 4% (6/143) and 14% (20/143) of the isolates, respectively. All bloodstream isolates were susceptible to voriconazole (MIC, < or = 1microgram/mL). CONCLUSION: Our findings show that C. albicans is the most common cause of Candida-related BSI, followed by C. parapsilosis, and that the rates of resistance to azole antifungals are still low among bloodstream isolates in Korea.
Antifungal Agents/*pharmacology ; Azoles/*pharmacology ; Bacteremia/*microbiology ; Candida/classification/*drug effects/isolation & purification ; Candidiasis/*microbiology ; Drug Resistance, Fungal ; Fluconazole/pharmacology ; Hospitals, University ; Humans ; Itraconazole/pharmacology ; Microbial Sensitivity Tests ; Population Surveillance ; Pyrimidines/pharmacology ; Triazoles/pharmacology