1.Characterization of Pseudallescheria boydii and Scedosporium apiospermum by Random Amplification of Polymorphic DNA Assay
Jingsi ZENG ; Fukushima KAZUTAKA ; Yuechen ZHENG ; Takizawa KAYOKO ; Nishimura KAZUKO
Chinese Journal of Dermatology 2003;0(08):-
Objective To investigate the DNA polymorphism of Pseudallescheria boydii and Scedosporium apiospermum and to analyze the relationship between random amplification of polymorphic DNA (RAPD) patterns and geographic origins.Methods The genomic DNAs of 13 Pseudallescheria boydii strains and 18 Scedosporium apiospermum strains isolated from 5 countries were amplified with RAPD technique.Results All strains tested were classified into 31 patterns by combination of the results obtained with 3 primers.The cluster tendency was identified based on species difference,namely,P.boydii or S.apiospermum strains,however,no such cluster tendency was revealed based on geographic origins of the most of strains,by dendrogram analysis.Conclusions The infraspecific variability of P.boydii and S.apiospermum is considerable.The cluster tendency of RAPD profiles is of consistency with morphological properties of P.boydii and S.apiospermum to some degree,however,is of no correlation with geographic origins of the pathogenic strains.
2.A DNA Genotyping Study of Candida Isolates from Mothers and Their Neonates
Liyan XI ; Fukushima KAZUTAKA ; Takizawa KAYOKO ; Changming LU ; Liqing CEN ; Nishimura KAZUKO
Chinese Journal of Dermatology 2003;0(08):-
Objectives To identify the route and time of transmission by Candida species from mothers' vagina to their neonates' mouth.Methods Specimens for fungal cultures were obtained from vaginal discharge of mothers just before delivery and also from the mouth of their offspring just after birth.Eleven mother-infant pairs were investigated.Candida species was identified based on morphology,biochemical analysis,and sequencing of D1/D2 domain of the large subunit of ribosomal DNA (LSUrDNA).Electrophoretic karyotyping (EK) and random amplified polymorphic DNA analysis (RAPD) were performed to search for DNA homology.Results Candida isolates (16 strains) from 8 mother-infant pairs were identified as Candida albicans by 100% homology of their D1/D2 sequences with reference strain C.albicans Y-12983 (GenBank access No.U45776).Similarly,4 strains from two mother-infant pairs and 2 isolates from the other pair were identified as Candida glabrata and Candida krusei,respectively,by 100% homology in sequences alignment of the domains with reference strains,C.glabrata Y-65(U44808) and C.krusei Y-5396 (U76347).The same EK profiles were found for each C.albicans or C.krusei strain pair from both mother and her neonate.Although different EK bands with various molecular size were generated for each C.glabrata isolate pair,they were still considered to be homologous based on the fact that main EK bands were identical.Each isolate pair from mother and her infant presented almost the same RAPD profile,except for one pair,isolates F7n and F7m,which showed minor diverse DNA bands.Conclusion Eleven Candida isolates from neonates have identical molecular characteristics with their mother's isolates.Vertical transmission may be the main pathway of Candida spp.from mothers to their neonates.
3.Identification of Trichophyton tonsurans by Random Amplified Polymorphic DNA.
Jeong Aee KIM ; Norma Buarque DE GUSMAO ; Kaoru OKADA ; Galba Maria DE CAMPOS TAKAKI ; Kazutaka FUKUSHIMA ; Kazuko NISHIMURA ; Makoto MIYAJI
Annals of Dermatology 1999;11(3):135-141
BACKGROUND: T. tonsurans is an anthropophilic dermatophyte mostly causing tinea capitis and tinea corporis. In East Asian countries, it has rarely been isolated until now. However, it is necessary for researchers in Asian countries to be more accustomed to T. tonsurans than before because of frequent international sports exchanges nowadays. OBJECTIVES: This study was performed to identify T. tonsurans by random amplified polymorphic DNA (RAPD) analysis. METHODS: Fifteen strains which were tentatively identified as T. tonsurans in Brazil were identified again by several conventional mycological tests and RAPD analysis. RESULTS: Among 15 Brazilian strains, 3 were identified as T. tonsurans, 8 T. mentagrophytes, 3 T. nJmwn and 1 T. raubitschekii by conventional mycological tests. This result was examined again by RAPD analysis. CONCLUSION: RAPD analysis is considered a rapid and reliable method for identification of T. tonsurans if the procedure is carefully standardized with adequate-primers.
Arthrodermataceae
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Asian Continental Ancestry Group
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Brazil
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DNA*
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Fungi
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Humans
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Methods
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Sports
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Tinea
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Tinea Capitis
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Trichophyton*
4.Identification of Trichosporon spp. strains by sequencing D1/D2 region and sub-typing by sequencing ribosomal intergenic spacer region of ribosomal DNA.
Jingsi, ZENG ; Cristina Maria, DE SOUZA MOTTA ; Kazutaka, FUKUSHIMA ; Kayoko, TAKIZAWA ; Oliane, MARIA CORREIA MAGALHES ; Rejane Pereira, NEVES ; Kazuko, NISHIMURA
Journal of Huazhong University of Science and Technology (Medical Sciences) 2009;29(5):655-8
To re-identify and further group 25 isolates of Trichosporon spp. identified morphologically previously, sequences of D1/D2 region of large subunit (LSU) of ribosomal DNA (rDNA) of 25 tested strains for identification and those of ribosomal intergenic space 1 (IGS1) region of 11 strains for subgrouping were detected. The identifications of tested strains were changed except 6 strains. According to the alignment of the IGS1 region, 6 T. asahii isolates tested fell into 4 groups and 5 T. faecale isolates into 3 groups. Polymorphism of 2 T. japonicum isolates was found in 10 positions. With the alignments obtained in this research compared with the relative GenBank entries, it was found that T. asahii, T. faecale and T. japonicum species were divided into 7, 3 and 2 subtypes respectively. Morphological and biophysical methods are not sufficient for Trichosporon spp. identification. Sequencing becomes necessary for Trichosporon diagnosis. There is obvious diversity within a species.
5.Identification of Trichosporon spp. Strains by Sequencing D1/D2 Region and Sub-typing by Sequencing Ribosomal Intergenic Spacer Region of Ribosomal DNA
ZENG JINGSI ; Cristina Maria de Souza Motta ; Fukushima KAZUTAKA ; Takizawa KAYOKO ; Oliane Maria Correia Magalhes ; Neves Pereira REJANE ; Nishimura KAZUKO
Journal of Huazhong University of Science and Technology (Medical Sciences) 2009;29(5):655-658
To re-identify and further group 25 isolates of Trichosporon spp. identified morphologically previously, sequences of D1/D2 region of large subunit (LSU) of ribosomal DNA (rDNA) of 25 tested strains for identification and those of ribosomal intergenic space 1 (IGS1) region of 11 strains for sub-grouping were detected. The identifications of tested strains were changed except 6 strains. According to the alignment of the IGS1 region, 6 T. asahii isolates tested fell into 4 groups and 5 T. faecale isolates into 3 groups. Polymorphism of 2 T.japonicum isolates was found in 10 positions. With the alignments obtained in this research compared with the relative GenBank entries, it was found that T. asahii, T.faecale and T.japonicum species were divided into 7, 3 and 2 subtypes respectively. Morphological and biophysical methods are not sufficient for Trichosporon spp. identification. Sequencing becomes neces-sary for Trichosporon diagnosis. There is obvious diversity within a species.