矢口 貴志

The species of the genus Emericella have been classified and identified on the basis of morphological features. However, the phylogenetic relationships in this genus have not been investigated. To clarify the relationships according to molecular phylogenetics, morphological characteristics, and growth temperature regimens in Emericella, multilocus sequencing analysis based on recent Aspergillus taxonomy was carried out. Various characteristic species formed individual clades, and maximum growth temperature reflected the phylogenetics. Emericella species exhibit various ascospore characteristics, although some species do not have distinct ascospore ornamentation. Species that have smooth-walled ascospores with two equatorial crests are polyphyletic. Here, Emericella pachycristata is described and illustrated as a new species. Its ascospores are similar to those of E. nidulans. These species produce smooth-walled ascospores, but the equatorial crests of E. pachycristata are thicker than those of E. nidulans. On the phylogenetic trees, E. pachycristata is closely related to E. rugulosa, which produces ascospores with ribbed convex surfaces. Thus, E. pachycristata is considered to be a new species both morphologically and phylogenetically.

Species of the genus Neosartorya are heat-resistant fungi that cause the spoilage of heat-processed acidic foods due to the formation of heat-resistant ascospores, and they produce mycotoxins, such as fumitremorgins and gliotoxin. Their anamorphs are phylogenetically and morphologically very close to Aspergillus fumigatus, which has never been reported as a spoilage agent in heat-processed food products. Therefore it is important to discriminate between the species of Neosartorya and A. fumigatus in the food industry. In the present study, we examined beta-tubulin and calmodulin genes to identify Neosartorya and A. fumigatus at the species level and found a region for specifically detecting these species. We succeeded in developing the PCR method of differentiating and identifying, Neosartorya and A. fumigatus using specific primer sets. Moreover, we developed specific primer sets to identify Neosartorya species, N. fischeri, N. glabra, N. hiratsukae, N. pseudofischeri, and N. spinosa-complex, which are important in food spoilage; these fungi vary in heat resistance and productivity of mycotoxins, depending on the species. PCR using these primer sets did not detect other fungi involved in food spoilage and environmental contamination. These identification methods are rapid and simple with extremely high specificity.

A three-year-old Sri Lankan boy residing in Japan developed a nodule on his scalp after visiting Sri Lanka. Two months later, the lesion increased in size to 22 x 19 mm(2), and was identified as an erythematous nodule with alopecia. Direct examination of the infected hair shafts indicated fungal hyphae outside the shafts. The fungus was identified as Microsporum gypseum following mycological examination. The sequence of the internal transcribed spacer 1 region of ribosomal RNA gene (ITS1 rDNA) exhibited 95.7-100.0% homology with that of Arthroderma incurvatum. The patient was successfully treated with a 6-week itraconazole course. We also examined DNA samples from eight clinical isolates of A. incurvatum. Alignments of ITS1 sequences of these strains and our isolate, showed gaps in the 64-bp positions 140-142 and 141-143 of the 205-207-bp ITS1 alignment. We performed phylogenetic analysis using the neighbor-jointing (NJ) method based on the ITS1 sequences of the present isolate and twenty related strains. Fifteen A. incurvatum strains were divided into East Asia and non-East Asia clusters. The present isolate belonged to the non-East Asia cluster, suggesting that the patient was infected outside Japan. Moreover, the trees suggested area-dependent genetic polymorphism of A. incurvatum

Aspergillus udagawae and A. fumigatus share similar morphological features but they differ genetically. There is also an important clinical distinction as A. udagawae is less sensitive to amphotericin B than A. fumigatus. We encountered a rare case of bronchial infection due to A. udagawae that was successfully treated with voriconazole. An 82-year-old woman with diabetes mellitus complained of bloody sputum. Bronchoscopy revealed a white plugged region at the origin of the right bronchi B5. Cytological study revealed a clot composed of filamentous fungi and Aspergillus spp. was detected by culture. Molecular analysis revealed that the causative agent was A. udagawae, and voriconazole was used for the treatment. In comparison to A. fumigatus, the A. udagawae strain isolated in this case was less sensitive to amphotericin B, less virulent in immunosuppressed mice, and more sensitive to hydrogen peroxide, features that are almost identical to those of the previously reported isolates of the fungus. We should be aware of the emergence of new Aspergillus species that might pose a clinical threat.

We report here a case of fungal keratitis caused by Aspergillus viridinutans, a member of the genus which was found to have clinical and antifungal susceptibility characteristics distinct from the species it most closely resembles, Aspergillus fumigatus. A 26-year-old woman presented with contact lens-associated keratitis, the etiologic agent of which was initially slow growing and found to be resistant to amphotericin B and voriconazole. Therapeutic keratoplasty was performed and an isolate from a corneal scraping was identified as A. viridinutans through partial DNA sequencing of the beta-tubulin gene, along with its slow growth 42 degrees C. In contrast with A. fumigatus, A. viridinutans proved to be resistant to a wider variety of antifungal agents. To the best of our knowledge, this is the first case of keratitis caused by A. viridinutans. Clinical isolates of the genus Aspergillus include rarely encountered species such as A. viridinutans. These species may show clinical manifestations distinct from those of A. fumigatus, including clinical resistance to amphotericin B and voriconazole and relatively low susceptibility to other antifungal agents.

Species of the genus Byssochlamys, heat-resistant fungi, cause the spoilage and poisoning of heat-processed acidic foods due to the formation of heat-resistant ascospores and the biosynthesis of patulin. Here we evaluated the heat resistance and patulin production of four species of Byssochlamys. Byssochlamys fulva was significantly more heat-resistant than the other species, and Byssochlamys lagunculariae and Byssochlamys nivea synthesized patulin. This is the first report of patulin production by B. lagunculariae. These findings indicate that the four species pose different health risks in food. Thus, we concluded that a method to identify the species of Byssochlamys is necessary. Identification at the species level based on morphology alone is difficult. We developed species-specific primers to identify B. nivea, B. fulva, B. lagunculariae, and Byssochlamys zollerniae using polymerase chain reaction (PCR) to amplify the beta-tubulin gene. PCR using the primer sets designed for B. nivea ( B. niveal F/1R) amplified PCR products specific for B. nivea. Similarly, PCR using the B.fulva1F/1R, B.lag1F/1R and B.zol3F/R primer sets produced PCR products specific for B. fulva, B. lagunculariae, and B. zollerniae, respectively. PCR using these four primer sets did not detect other fungi involved in food spoilage and environmental contamination. This identification method is rapid and simple with extremely high specificity. (C) 2012 Elsevier Ltd. All rights reserved.

64歳女性。半年前より右頬部にそう痒を伴う皮疹が出現し、クロベタゾールプロピオン酸エステルを外用するも改善せず、著者らの施設へ受診となった。所見では右頬部に鱗屑と痂皮が付着し、隆起を伴う境界明瞭な浸潤性の紅斑局面が認められた。また、病理組織学的にはPAS染色やグロコット染色にて多数の菌糸がみられ、真菌培養では黒色真菌が検出された。以上、これらを踏まえ、遺伝子検査を行った結果、本症例はOchroconis humicola感染症と診断され、真菌による内臓病変がないことからイトラコナゾールの内服を開始した。だが、胃部不快感が出現したためテルビナフィンに変更したところ、1ヵ月で皮疹は徐々に改善した。その後、8ヵ月間はテルビナフィンの内服が継続されたが、内服終了後10ヵ月経過した現在、再燃はみられていない。

To report the predisposing factors, patient background, treatment and outcomes of ocular infections caused by Paecilomyces spp. in Japan. We reviewed the medical records and the published literature in Japan on ocular infections caused by microbiologically-proven Paecilomyces spp. Nineteen eyes of 18 patients; 4 eyes of 4 of our patients and 15 eyes of 14 published Japanese patients were studied. Ten men and 8 women (9 OD, 8 OS, 1 OU) were diagnosed with ocular infections caused by Paecilomyces spp. The mean age was 69 years (range 33 to 90 years). The initial diagnosis of ocular infection caused by Paecilomyces spp. was keratitis in 14 eyes and endophthalmitis in 5 eyes. The final diagnosis was keratitis in 2 eyes, keratitis with involvement of the anterior segment of the eye, i.e., hypopyon or corneal rupture, in 12 eyes, and endophthalmitis in 5 eyes. The predisposing factors had a history of ocular surgery, corneal trauma and soft contact lens use. Other factors were diabetes and old age. The infections developed more often in the fall and winter. The identified Paecilomyces spp. had a high sensitivity to micafungin and voriconazole, but low sensitivity to amphotericin B, flucytosine and fluconazole. The prognosis of Paecilomyces spp. ocular infections was very poor, viz., final vision was counting fingers or worse in 60%, ocular perforation in 42%, and eye enucleation in 11%. The percentage of eyes with a final visual acuity worse than counting fingers was 0% in those with keratitis, 50% in those with involvement of the anterior segment, and 100% in those with endophtalmitis (p = 0.0446). Among the antifungal agents, the percentage of cases with final vision of counting fingers or worse was 90% with fluconazole, 80% with itraconazole, 100% with miconazole, and 71% with voriconazole. The results show that if the Paecilomyces spp. infection spreads intraocularly, it is difficult to mitigate the clinical damage even with antifungal agents with high sensitivity.

Genotype characteristics and distribution of commensal Candida albicans should be studied to predict the development of candidiasis, however, extensive genotype analysis of commensal C. albicans has not been made. In this study, 508 C. albicans isolates were collected from patients with/without candidiasis and divided into 4 isolate groups (SG-1, oral cavity of non-candidiasis patients SG-2, patients with cutaneous candidiasis SG-3, patients with vaginal candidiasis SG-4, patients with candidemia). These isolates were characterized to study the relationship between genotypes and pathogenicity using microsatellite analysis. Using CDC3 and CAI, 5 genotypes (I, 111: 115/33: 41 II, 115: 119/23: 23 III, 115: 123/18: 27 IV, 115: 123/33: 40 and V, 123: 127/32: 41) were found in 4.2%, 8.9%, 7.1%, 2.2% and 3.1% of the isolates, respectively. Genotypes II and III were commonly found in all isolate groups. These genotypes were further divided into 28types by additional HIS3 and CAIII microsatellite markers. In this analysis, C. albicans with type 6 and type 23 was widely distributed as a commensal species in the oral cavity of non-candidiasis patients and found to be related with candidiasis development. Additionally, genotypes I and IV were found in SG-2 and/or SG-4, suggesting that the fungus with those genotypes is also involved in this development. In contrast, genotype V was not identified in any infective isolates.

Genotype characteristics and distribution of commensal Candida albicans should be studied to predict the development of candidiasis, however, extensive genotype analysis of commensal C. albicans has not been made. In this study, 508 C. albicans isolates were collected from patients with/without candidiasis and divided into 4 isolate groups (SG-1, oral cavity of non-candidiasis patients; SG-2, patients with cutaneous candidiasis; SG-3, patients with vaginal candidiasis; SG-4, patients with candidemia). These isolates were characterized to study the relationship between genotypes and pathogenicity using microsatellite analysis. Using CDC3 and CAI, 5 genotypes (I, 111: 115/33: 41; II, 115: 119/23: 23; III, 115: 123/18: 27; IV, 115: 123/33: 40; and V, 123: 127/32: 41) were found in 4.2%, 8.9%, 7.1%, 2.2% and 3.1% of the isolates, respectively. Genotypes II and III were commonly found in all isolate groups. These genotypes were further divided into 28 types by additional HIS3 and CAIII microsatellite markers. In this analysis, C. albicans with type 6 and type 23 was widely distributed as a commensal species in the oral cavity of non-candidiasis patients and found to be related with candidiasis development. Additionally, genotypes I and IV were found in SG-2 and/or SG-4, suggesting that the fungus with those genotypes is also involved in this development. In contrast, genotype V was not identified in any infective isolates.

Two new indole diterpenes, 8,21-dehydro-17-hydroxyeujindole (3) and 8,21-dehydro-17,20-epoxyeujindole (4) were isolated from the extract of Eupenicillium javanieum IFM 59075. The structures were determined by spectroscopic methods.

本菌の活性本体は常に大量のスピクリスポール酸（SPA）（3） と挙動を共にするが、3は抗A. fumigatus活性を示さなかった。各種カラムクロマトグラフィーにて活性本体の分取を試みるが、顕著な活性の低下をもたらした。<br> 活性分画から3を再結晶で出来るだけ除去後、ジアゾメタンで処理し、2-（2-カルボキシエチル）-3-デシルマレイン酸無水物モノメチルエステル（2）、SPAジメチルエステル（4）、2-（2-カルボキシエチル）-3-デシル-2-ヒドロキシコハク酸トリメチルエステル（6）を得た。よって、活性本体は2または6の脱メチル体であることが分かった。1と5を調整し活性を確認したところ、1が活性を示した。また、活性を示す分画の¹H NMRスペクトルには化合物1の特徴的なシグナルを確認できたことを併せ、本菌の活性本体を化合物1と決定した。

神経ブロック注射部位に生じた皮膚アスペルギルス症の1例を経験したので報告する。67歳女。6年前より成人スティル病、間質性肺炎でプレドニン、免疫抑制剤内服治療中であった。帯状疱疹後神経痛に対して某ペインクリニックで左腰部に神経ブロック注射を施行。その後、注射部位に一致して出現した軽度圧痛のある3つの皮下結節を主訴に当科を紹介受診。1つを切除生検。術中、結節内部より緑色の膿汁の排出を認めた。病理組織学的には皮下組織に著明な好中球浸潤があり、膿瘍を形成していた。PAS染色、Grocott染色に染まる菌糸を認め、組織、膿汁の真菌培養にてAspergillus spp.が分離され、全身検索では内臓病変なし。原発性皮膚アスペルギルス症と診断した。残りの2つを全摘の上、イトラコナゾールの内服治療を開始するも皮下膿瘍が再発。分離培養された真菌のリポゾームRNA遺伝子のITS領域の塩基配列の解析でAspergillus ustusと同定し、その後、37℃での発育およびβ-tubulin遺伝子の塩基配列によりA.calidoustusと同定した。文献上のA.ustusおよびA.calidoustusの薬剤感受性に従い、テルビナフィン内服に変更しデブリードマンを予定していたが、間質性肺炎の急性増悪のために永眠された。神経ブロック部位に生じた原発性皮膚アスペルギルス症はまれであるが、免疫不全患者では十分な注意が必要である。(著者抄録)

We report a lymphocutaneous type of nocardiosis caused by Nocardia vinacea. A 62-year-old woman with polymyositis presented with some erythematous swellings and subcutaneous abscesses on her right middle finger and the dorsum of her hand, which had persisted for 2 weeks. Culturing of the excised nodule and pus revealed orange to orange-tan colonies with scanty whitish aerial mycelia. The isolate was identified as N. vinacea on the basis of its biochemical and chemotaxonomic characteristics and the results of molecular biological analysis. In our case, oral minocycline (MINO) and trimethoprim-sulfamethoxazole (TMP-SMX) for 7 weeks did not improve the clinical manifestation, even though in vitro susceptibility testing of the isolate predicted its susceptibility to MINO and TMP-SMX. Treatment with partial surgical excision followed by TMP-SMX and meropenem administration was effective. This is the first reported case of a lymphocutaneous type of nocardiosis caused by N. vinacea.

A Gram-staining-positive bacterium, designated AS-0823(T), which formed spiral spore chains on the aerial mycelium, was isolated from the intestinal tract of Armadillidium vulgare, a small terrestrial crustacean commonly found on the ground around houses in Japan. 16S rRNA gene sequence analysis showed that the isolate belonged to the genus Streptomyces and was most closely related to Streptomyces longisporus ISP 5166(T) (98.6% 16S rRNA gene sequence similarity), Streptomyces curacoi NBRC 12761(T) (98.4%) and Streptomyces griseoruber NBRC 12873(T) (98.4%). The affiliation of strain AS-0823(T) to the genus Streptomyces was supported by chemotaxonomic data: iso-C(16:0), anteiso-C(15:0), C(16:0), iso(15:0) and anteiso-C(17:0) as the major cellular fatty acids, LL-diaminopimelic acid as the characteristic diamino acid in the peptidoglycan and the absence of mycolic acids. DNA DNA hybridization and physiological and biochemical analysis supported the differentiation of strain AS-0823(T) from S. longisporus JCM 4395(T). Therefore, strain AS-0823(T) represents a novel species, for which the name Streptomyces coacervatus sp. nov. is proposed. The type strain is AS-0823(T) (=IFM 11055(T) = DSM 41983(T) = JCM 17138(T)).

We describe a disseminated fungal infection by Conidiobolus lamprauges in a patient with malignant lymphoma. Histopathology and mycological studies were performed, along with molecular analyses. This is the first record of this species causing human disease and the fifth reported disseminated infection by a Conidiobolus sp. in humans.

Two new indole diterpenes, 17-hydroxyeujindole (1) and 17-oxoeujindole (2), were isolated from the extract of Eupenicillium javanicum IFM 59075 (UC62). The structures containing the absolute configuration were determined by spectroscopic methods.

Owner : NLM<br /> Status : In-Process<br /> PubModel : Print<br /> Language : eng<br /> Pagination : 129-38

This study aimed to examine the genotype distribution of Candida albicans and the major genotypes involved in superficial candidiasis. The genotypes of C. albicans isolated from the infection sites of patients with superficial candidiasis(referred to as infection isolates) were analyzed by fragment analysis using 4microsatellite markers(HIS3, CDC3, CAI and CAIII). Genotypes of the infection isolates were compared with those of C. albicans isolated from oral mucosa of non-candidiasis patients (referred to as oral isolates). Isolates of C. albicans showed 4major genotypes for HIS3/CAI(,a-for 148: 148/23: 23, "b"-for 148: 160/33: 41, "c"-for 148: 164/32: 41 and, "d"-for 152: 152/18: 27). The genotypes, "a", "b" and, "d" were commonly found in oral (4.7, 8.8 and 7.6%, respectively) and infection (6.6, 9.2 and 15.4%, respectively) isolates. No isolates of genotype, "c" were isolated from infection sites. The genotype, "a" was found in the isolates from patients with genitalia candidiasis. Genotyping of multiple isolates from an individual patient showed that C. albicans from infection sites was genetically homogenous as compared with that of oral isolates, even in the same patient with candidiasis.

We report a case of primary cutaneous Aspergillus caldioustus infection caused by nerve block therapy. A 67-year-old Japanese woman had been treated with oral predonisolon and tacrolimus for adult-onset Still disease and interstitial pneumonia. She presented with a 2-month-history of the lesions on the left back. A biopsy specimen from the skin lesion revealed granulomatous inflammation with hyphae. Culture of the pus and the skin specimen confirmed the diagnosis of cutaneous Aspergillus infection. The sequence of β- tubulin gene was analyzed to confirm the mycological diagnosis and the causative agent was identified as A. caldioustus. The patient was treated with surgical removal of the lesions and oral 200 mg/day itraconazole but she died of infectious interstitial pneumonia due to Pneumocystis jiroveci and Cytomegalovirus infection Percutaneous infection may have been responsible for the incidence of localized infection. There was no evidence of systemic aspergillosis. A. caldioustus is an emerging opportunistic fungal pathogen in immunocompromised patients. Immunocompromised patients who have persistent traumatic atypical skin lesion need to be ruled out of such rare fungus infection. An opportunistic infection in Immunocompromised patients can be life-threatening and prompt treatment based on accurate diagnosis is important.

Fonsecaea species are major etiologic agents of Chromoblastomycosis(CBM). By genetic analysis, the genus Fonsecaea has recently been revised and classified into F. pedorosoi, F. monophora and F. nubica. Here we report a severe chronic case of CBM caused by F. monophora. A 55-year-old Filipino male developed progressive skin lesions on the left lateral ankle in 1973, when he worked at a coconut plantation in the Philippines. In 1999, he received medical treatments for enlarged, multiple lesions on the left lower limb. When he moved to Japan in 2005, the lesions were remarkably improved and he discontinued taking the medicine. On our first examination in October 2008, a large, reddish, cicatricial plaque was observed on the left lower aspect of his leg. Several tumorous lesions surrounded the plaque, indicating that the therapies performed before had been insufficient. In addition, there were many patchy scars scattered on the thigh and the upper part of the lower leg. The diagnosis of CBM was made by the presence of muriform cells. Black, pulverulent colonies were yielded in culture of skin scrapings and tissues. Although the fungus could not be identified by microscopic morphology, r-RNA ITS sequence analysis enabled identification of Fonsecaea monophora. The patient responded well to oral voriconazole combined with local thermotherapy using pocket warmers. The tumoral masses subsided in 6 months, leaving pink scars with negative fungal culture. Voriconazole treatment was continued for 18 months. It seems that drugs are insufficiently delivered in the cicatricial lesions because of the paucity of blood flow, suggesting that a long-term follow-up is necessary for such a case.

Three new quinazolinobenzodiazepine derivatives, novobenzomalvins A (1), B (2), and C (3), have been isolated as fibronectin expression regulators from Aspergillus novofumigatus CBS117520. The structures of 1 to 3 were established by spectroscopic and physicochemical analysis, and chemical investigation including the total synthesis of 1. Treatment with novo-benzomalvins A (1), B (2), C (3), and N-methylnovobenzomalvin A (5) increased the expression of fibronectin in normal human neonatal dermal fibroblast cells. © Ishikawa et al.

A new cyclic tripeptide, novofumigatamide (1) has been isolated from Aspergillus novofumigatus CBS117520. The relative structure of I was established on the basis of spectroscopic and physico-chemical data and chemical investigations. The absolute structure of 1 was determined by using a Marfey&apos;s method. Compound I showed non-specific antifungal activities against some human pathogenic fungi and did not inhibit cell proliferation for any tumor cells.

Two new species, Neosartorya shendaweii and N. tsunodae, isolated from soil in Xinjing, China and in Pernambuco, Brazil, are described and illustrated. The first species is characterized by its ascospores with two widely separated equatorial crests and tuberculate to verrucose convex surfaces. This species has affinities with several known species of the genus, bearing ascospores with a similar ornamentation, but can be distinguished from these species by other morphological characteristics such as smaller cleistothecia and conidiophores, spathulate vesicles and rather ellipsoidal conidia. The second species is characterized by its unique ascospores with two low equatorial crests, an evident furrow as a deep depression, and finely reticulate convex surfaces. The validation of these new species is supported further by analyses of the beta-tubulin, calmodulin and actin gene sequences.

Two new diketopiperazine metabolites, novoamauromine (1) and ent-cycloechinulin (2) have been isolated from Aspergillus novofumigatus CBS117520. The structures of 1 and 2 were established on the basis of spectroscopic and chemical investigation, including a detailed comparison of the spectroscopic and physico-chemical data of amauromine (3) and cycloechinulin (4).

Owner : NLM<br /> Status : MEDLINE<br /> PubModel : Print<br /> Language : eng<br /> Pagination : 109-16

Antifungal susceptibility tests were performed for 38 Candida albicans strains isolated from oral cavities of 43 Uighurian AIDS patients. Results showed that six isolates were resistant to fluconazole; one showed low susceptibility. We attempted to examine these strains molecular-epidemiologically, but 25S rDNA genotyping was insufficient for their discrimination. To estimate whether the origins of resistant strains were identical, we developed a new combination method of C. albicans tandem repeating units (ALTS)/RFLP and micro-temperature gradient gel electrophoresis (micro-TGGE). This new method was able to distinguish all seven strains. A suspected nosocomial infection was ruled out.

Heat-resistant fungi, genera Byssochlamys, Talaromyces, Neosartorya, and Hamigera, contribute significantly to the spoilage of heat-processed acidic foods, due to the formation of heat-resistant ascospores. Here, we first evaluated the differences in the β-tubulin gene between Byssochlamys and Hamigera and developed specific primers to identify the Byssochlamys species fulva, nivea, and spectabilis, and Hamigera. Using primers designed for B. fulva and B. nivea (B1F/1R), specific PCRr products were detected for B. fulva and B. nivea, as well as B, langunculariae and B. zollerniae, two closely related species. Similarly, the Pae4F/4R-1 and H2F/2R primers produced specific PCRr products for B. spectabilis and Hamigera, respectively. Using these three primer sets, strains involved in acidic food spoilage and environmental contamination were not detected. The detection limits of all primer sets were 1 ng of DNA by PCRr and 10 pg of DNA by nested PCRr. Each PCRr assay was specific, even if the sample was contaminated 1,000-fold by other fungal DNA. Thus, this method has proved to possess an extremely high degree of specificity. Copyright © International Association for Food Protection.

Aspergillosis is an important mycosis caused primarily by Aspergillus fumigatus and its relatives. The genus Emericella is a teleomorph related to the Aspergillus section Nidulantes. The typical anamorphic stage species in this genus is Aspergillus nidulans, which is sometimes a significant agent in chronic granulomatous disease (CGD) patients. The mortality rate of osteomyelitis in CGD patients due to A. nidulans (E. nidulans) is very high compared to that due to A. fumigatus. Moreover, two Emericella species (E. nidulans and E. quadrilineata) from clinical specimens exhibit different sensitivities against several antifungal drugs. In aspergillosis, correct species identification is important for antifungal therapy. We attempted to develop rapid and specific molecular discrimination by polymerase chain reaction (PCR) and loop-mediated isothermal amplification (LAMP) methods in the principal pathogenic Emericella species, and succeeded in establishing species-specific primers corresponding to the hydrophobin gene. These primers discriminate E. nidulans and E. quadrilineata rapidly and specifically. These methods and primers make it possible to diagnose etiological agents in aspergillosis quickly and easily.

Antifungal susceptibility tests were performed for 38 Candida albicans strains isolated from oral cavities of 43 Uighurian AIDS patients. Results showed that six isolates were resistant to fluconazole; one showed low susceptibility. We attempted to examine these strains molecular-epidemiologically, but 25S rDNA genotyping was insufficient for their discrimination. To estimate whether the origins of resistant strains were identical, we developed a new combination method of C. albicans tandem repeating units (ALTS)/RFLP and micro-temperature gradient gel electrophoresis (micro-TGGE). This new method was able to distinguish all seven strains. A suspected nosocomial infection was ruled out.