A New Isolate of the Genus
Malassezia
Based
on the Sequence Analysis of 26S and ITS1 in
Ribosomal DNA
Hossein Mirhendi, Ph.D.1*, Ali Zia, Ph.D.2, Kuichi Makimura, M.D., Ph.D.3
1. Medical Parasitology and Mycology Department, School of Public Health and Institute of Public Health Researches, Tehran University of Medical Sciences, Tehran, IRAN
2. Azad University, Khorasgan, Isfahan, Iran
3. Institute of Medical Mycology and Genome Research Center, Teikyo University, Tokyo, Japan * Corresponding Address: P.O.Box: 14155-6446, Medical Parasitology and Mycology Department, School of Public Health
and Institute of Public Health Researches, Tehran University of Medical Sciences, Tehran, IRAN Email: mirhendi@tums.ac.ir
Received: 21/May/2008, Accepted: 12/Nov/2008 Abstract
Malassezia species considered to be the etiological agents of pityriasis versicolor and Malassezia follicolitis in humans. Recently, on the basis of molecular data, four new spe-cies were added to the genus. In total, 11 spespe-cies have been described and accepted so far. In this study we describe a new isolate of Malassezia based on the nucleotide se-quence of 26SrDNA and ITS1 regions, as the accepted critical markers for description of the species.
The yeast was isolated from a hamster. Two primer pairs, one for amplification of D1/D2-26Sr DNA and another for the ITS1 region were used in PCR. The PCR products were sequenced and analyzed to compare with other similar sequences which are already de-posited in the GenBank. The 26SrDNA PCR product was also digested with the restriction enzyme CfoI.
Malassezia-specific universal primer pairs successfully amplified the 26srDNA and ITS1 regions of the new isolate, providing a single PCR product of about 580 and 280 base pairs, respectively. After digestion of the 26s PCR product with the enzyme CfoI, a unique and different RFLP pattern was observed. Sequence analysis of D1/D226s and ITS1 re-gions were compared with the same rere-gions in all already described Malassezia species, which implied a different and unique new sequences. The phylogenetic tree of both regions showed that the isolate could be a different Malassezia isolate.
Regarding the new RFLP pattern of D1/D226SrDBA and the unique nucleotide sequence of both D1/D2 26SrDNA and ITS1 regions, we propose the isolate to be a new Malassezia.
Keywords: Malassezia, Ribosomal DNA, Classification
(
Malassezia
)
ﺎﻳﺰﺳﻻﺎﻣ
ﺲﻨﺟ
ﻱﺮﻤﺨﻣ
ﻱﺎﻫ
چﺭﺎﻗ
ﺯﺍ
ﻱﺪﻳﺪﺟ
ﻪﻟﻭﺰﻳﺍ
ﻲﻓﺮﻌﻣ
ﻲﻣﻭﺯﻮﺒﻳﺭ
ﻱ
DNA
ﺭﺩ
ITS1
ﻭ
26S
ﻪﻴﺣﺎﻧ
ﻱﺎﻫﺪﻴﺗﻮﺌﻠﻛﻮﻧ
ﻲﻟﺍﻮﺗ
ﺰﻴﻟﺎﻧﺁ
ﺱﺎﺳﺍ
ﺮﺑ
3 ،،
Ph.D.،،M.D.ﺍﺭﻮﻣﻲﻛﺎﻣﻲﭽﻴﺋﻮﻛ،
2 ،،
Ph.D.ﺎﻴﺿﻲﻠﻋﺪﻤﺤﻣ،*1
،،Ph.D.ﻱﺪﻨﻫﺮﻴﻣﻦﻴﺴﺣﺪﻴﺳ
ﻥﺍﺮﻳﺍ،ﻥﺍﺮﻬﺗ،ﻲﺳﺎﻨﺷچﺭﺎﻗﻭﻲﺳﺎﻨﺷﻞﮕﻧﺍﻩﻭﺮﮔ،ﺖﺷﺍﺪﻬﺑﻩﺪﻜﺸﻧﺍﺩ،ﻥﺍﺮﻬﺗﻲﻜﺷﺰﭘﻡﻮﻠﻋﻩﺎﮕﺸﻧﺍﺩ .1
ﻥﺍﺮﻳﺍ،ﻥﺎﻬﻔﺻﺍ،ﻥﺎﮕﺳﺍﺭﻮﺧﺪﺣﺍﻭ،ﻲﻣﻼﺳﺍﺩﺍﺯﺁﻩﺎﮕﺸﻧﺍﺩ .2
ﻦﭘﺍژ،ﻮﻴﻛﻮﺗ،ﻮﻴﻜﻴﺗﻩﺎﮕﺸﻧﺍﺩﻲﻜﺷﺰﭘﻲﺳﺎﻨﺷچﺭﺎﻗﻮﺘﻴﺘﺴﻧﺍ .3
ﻲﺳﺎﻨﺷچﺭﺎﻗﻭﻲﺳﺎﻨﺷﻞﮕﻧﺍﻩﻭﺮﮔ،ﺖﺷﺍﺪﻬﺑﻩﺪﻜﺸﻧﺍﺩ،ﻥﺍﺮﻬﺗﻲﻜﺷﺰﭘﻡﻮﻠﻋﻩﺎﮕﺸﻧﺍﺩ،14155-6446 :ﻲﺘﺴﭘﻕﻭﺪﻨﺻ،ﻥﺍﺮﻳﺍ،ﻥﺍﺮﻬﺗ :ﻝﻮﺌﺴﻣﻩﺪﻨﺴﻳﻮﻧﺱﺭﺩﺁ *
Email:mirhendi@tums.ac.ir:ﻚﻴﻧﻭﺮﺘﻜﻟﺍﺖﺴﭘ
87/8/22 :ﻪﻟﺎﻘﻣﺵﺮﻳﺬﭘ،87/3/1 :ﻪﻟﺎﻘﻣﺖﻓﺎﻳﺭﺩ
ﻩﺪﻴﻜﭼ
ﻲﻳﺎﻫﺮﻛﺭﺎﻣﻪﻛITS1ﻪﻴﺣﺎﻧﻲﻟﺍﻮﺗﻪﻛD1/D2 26S rDNAﻪﻴﺣﺎﻧﻚﻴﺌﻠﻛﻮﻧﻱﺎﻫﺪﻴﺳﺍﻲﻟﺍﻮﺗﺱﺎﺳﺍﺮﺑﺮﺿﺎﺣﺵﺭﺍﺰﮔ
.ﺪﻳﺎﻤﻧﻰﻣﻰﻓﺮﻌﻣﺍﺭﺲﻨﺟﻦﻳﺍﺯﺍﻱﺪﻳﺪﺟﻪﻟﻭﺰﻳﺍ،ﺖﺳﺎﻫﻪﻧﻮﮔﻕﺍﺮﺘﻓﺍﻱﺍﺮﺑ
D1/ﻪﻌﻄﻗﺖﻬﺟﻲﻜﻳ،ﺮﻤﻳﺍﺮﭘﺖﻔﺟﻭﺩﺯﺍﻩﺩﺎﻔﺘﺳﺍﺎﺑ .ﺪﻳﺩﺮﮔﻯﺯﺎﺳﺍﺪﺟﻝﻮﻬﺠﻣﻯﺎﻳﺰﺳﻻﺎﻣﻰﻧﻮﻠﻛﻭﺪﺷ ﻩﺩﺍﺩﺖﺸﻛﻪﻧﻮﻤﻧ
ﺕﺎﻌﻄﻗﻯﺪﻴﺗﻮﺌﻠﻛﻮﻧﻰﻟﺍﻮﺗﻭﻩﺪﺷﻡﺎﺠﻧﺍPCR،ﻲﻣﻭﺯﻮﺒﻳﺭﻱDNAﻪﺑﻁﻮﺑﺮﻣTS1ﻪﻴﺣﺎﻧﻯﺮﮕﻳﺩﻭ26SﻪﻴﺣﺎﻧﺭﺩﺩﻮﺟﻮﻣD2
ﺎﻫﻢﺴﻴﻧﺎﮔﺭﺍﻭﺮﻜﻴﻣﺮﻳﺎﺳﻭﺎﻫﺎﻳﺰﺳﻻﺎﻣﺮﻳﺎﺳﺭﺩﻰﺣﺍﻮﻧﻥﺎﻤﻫﻯﺎﻫﻰﻟﺍﻮﺗﺎﺑﻭﻪﺘﻓﺮﮔﺭﺍﺮﻗ ﺰﻴﻟﺎﻧﺁﺩﺭﻮﻣﺎﻫﻲﻟﺍﻮﺗ .ﺪﻳﺩﺮﮔﻦﻴﻴﻌﺗﺭﻮﺑﺰﻣ .ﺪﺷﻪﺴﻳﺎﻘﻣ
ﺎﻳﺰﺳﻻﺎﻣﺲﻨﺟﺯﺍﻲﻳﺎﻫﺮﻤﺨﻣﺩﻮﺟﻭﺯﺍﺖﻳﺎﻜﺣﻱﺮﻤﺨﻣﻱﺎﻫﻝﻮﻠﺳﻲﭘﻮﻜﺳﻭﺮﻜﻴﻣﻱژﻮﻟﻮﻓﺭﻮﻣﺰﻴﻧﻭﻪﺘﻓﺎﻳﺪﺷﺭﻱﺎﻫﻲﻧﻮﻠﻛ
.ﺪﻳﺩﺮﮔﻩﺪﻫﺎﺸﻣﺎﻫﺎﻳﺰﺳﻻﺎﻣﺮﻳﺎﺳﺎﺑﻪﺑﺎﺸﻣﻭﺯﺎﺑﺖﻔﺟ 580 ﺩﻭﺪﺣﻥﺯﻭﻪﺑDNAﻪﻌﻄﻗﻚﻳPCRﺶﻨﻛﺍﻭﻱﺍﺮﺟﺍﺎﺑ .ﺖﺷﺍﺩ
ﻱﺎﻫﺎﻳﺰﺳﻻﺎﻣﺯﺍﻡﺍﺪﻛﭻﻴﻫﺎﺑﻪﻛﺪﻣﺁﺖﺳﺩﻪﺑﻰﻳﻮﮕﻟﺍ،RFLPﺕﻻﻮﺼﺤﻣﺯﺭﻮﻓﻭﺮﺘﻜﻟﺍﻭ،CfoIﻢﻳﺰﻧﺁﺎﺑPCRﻝﻮﺼﺤﻣﺵﺮﺑ
ﻩﺪﻫﺎﺸﻣ ﺯﺎﺑﺖﻔﺟ 280ﺩﻭﺪﺣﻥﺯﻭﻪﺑﺪﻧﺎﺑﻚﻳﺰﻴﻧITS1ﻪﺑﻁﻮﺑﺮﻣPCRﻝﻮﺼﺤﻣﺯﺭﻮﻓﻭﺮﺘﻜﻟﺍﺎﺑ .ﺖﺷﺍﺪﻧﺖﻘﺑﺎﻄﻣﻩﺪﺷﻪﺘﺧﺎﻨﺷ
ﻲﻬﺟﻮﺗﻞﺑﺎﻗﻥﺍﺰﻴﻣﻪﺑﺎﻫﺎﻳﺰﺳﻻﺎﻣﺮﻳﺎﺳﺎﺑﻪﻟﻭﺰﻳﺍﻦﻳﺍﻯﺎﻫﺲﻧﺎﻜﺳﻪﻛﺪﺷﺺﺨﺸﻣ ITS1ﻭ26Sﻱﺎﻫﻰﻟﺍﻮﺗﺰﻴﻟﺎﻧﺁﺯﺍﺲﭘ .ﺪﻳﺩﺮﮔ .ﺩﺭﺍﺩﺕﻭﺎﻔﺗ
ﺰﻴﻧﻭD1/D2ﻲﺣﺍﻮﻧﻪﻜﻨﻳﺍﻪﺑﺮﻈﻧﻭﺪﺷﻩﺪﻫﺎﺸﻣ26S rDNAﻪﻴﺣﺎﻧRFLPﺯﺍﻞﺻﺎﺣﺞﻳﺎﺘﻧﺭﺩﻪﻛﻱﺪﻳﺪﺟﻱﻮﮕﻟﺍﻪﺑﻪﺟﻮﺗﺎﺑ
ﻭﺪﻨﺷﺎﺑﻲﻣﺎﻫﺎﻳﺰﺳﻻﺎﻣﻚﻴﻣﻮﻧﻮﺴﻛﺎﺗﻩﺎﮕﻳﺎﺟﻦﻴﻴﻌﺗﻱﺍﺮﺑﻯﺍﻩﺪﺷﻪﺘﻓﺮﻳﺬﭘﻱﺎﻫﺮﻛﺭﺎﻣ rDNAﻰﻧژﺲﻜﻠﭙﻤﻛﺭﺩﺩﻮﺟﻮﻣ ITS1
ﺎﻫﺎﻳﺰﺳﻻﺎﻣﺮﻳﺎﺳﺎﺑﺕﻭﺎﻔﺘﻣﻼﻣﺎﻛﻯﺪﻴﺗﻮﺌﻠﻛﻮﻧﻰﻟﺍﻮﺗﻯﺍﺭﺍﺩﻖﻴﻘﺤﺗﻦﻳﺍﻰﻃﺭﺩﻩﺪﺷﺍﺪﺟﻱﺎﻳﺰﺳﻻﺎﻣﺭﺩﻰﺣﺍﻮﻧﻦﻳﺍﻪﻛﺎﺠﻧﺁﺯﺍ
.ﺩﻮﺷﻲﻣﻲﻓﺮﻌﻣﺪﻳﺪﺟﻚﻴﻣﻮﻧﻮﺴﻛﺎﺗﻭﻰﻜﻴﺘﻧژﺕﺎﺼﺨﺸﻣﺎﺑﻪﻟﻭﺰﻳﺍﻚﻳﻥﺍﻮﻨﻋﻪﺑ ،ﺩﻮﺑ
ﻱﺪﻨﺑﻪﻘﺒﻃ،ﻲﻣﻭﺯﻮﺒﻳﺭDNA،ﺎﻳﺰﺳﻻﺎﻣ:ﻥﺎﮔژﺍﻭﺪﻴﻠﻛ *
244-249 :ﺕﺎﺤﻔﺻ،88 ﻥﺎﺘﺴﺑﺎﺗ،2 ﻩﺭﺎﻤﺷ،ﻢﻫﺩﺯﺎﻳﻝﺎﺳ،ﻪﺘﺧﺎﻳﻰﻜﺷﺰﭘﻪﻣﺎﻨﻠﺼﻓ
ﻪﻣﺪﻘﻣ
ﺭﻮﻠﻓ (Malassezia) ﺎﻳﺰﺳﻻﺎﻣ ﺲﻨﺟ ﻪﺑ ﻪﺘﺴﺑﺍﻭ ﻱﺎﻫﺮﻤﺨﻣ
ﺕﺎﻴﺻﻮﺼﺧ ﺯﺍ .ﺪﺷﺎﺑﻲﻣ ﻡﺮﮔﻥﻮﺧ ﺕﺎﻧﺍﻮﻴﺣ ﻭ ﻥﺎﺴﻧﺍ ﺖﺳﻮﭘ ﻲﻌﻴﺒﻃ
ﺎﻬﻧﺁ ﺮﻴﺜﻜﺗ ﻭ ﺪﺷﺭ ﻪﻣﺯﻻ ﻲﺑﺮﭼ ﻪﺑ ﺯﺎﻴﻧ ﺎﻫﺮﻤﺨﻣ ﻦﻳﺍ ﻩﺪﻤﻋ
ﻱﺩﺎﻋ ﻝﺎﺣ ﺭﺩ ﺎﻫﺎﻳﺰﺳﻻﺎﻣ .(1) ﺖﺳﺍ (Lipophilic Yeasts)
ﺖﻟﺎﺣ ﻲﺻﺎﺧ ﻂﻳﺍﺮﺷ ﺭﺩ ﻲﻟﻭ ﻩﺩﻮﺑ ﺖﺳﻮﭘ ﺭﺍﺯﺁ ﻲﺑ ﻱﺎﻫﻝﺎﺴﻨﻣﻮﻛ ﻲﺷﺎﻧﻱﺭﺎﻤﻴﺑﻦﻳﺮﺗﺭﻮﻬﺸﻣ .ﺩﻮﺷﻲﻣﺍﺯﻱﺭﺎﻤﻴﺑﻭﻪﺘﻓﺮﮔﺩﻮﺧﻪﺑﻲﻤﺟﺎﻬﺗ
ﺮﻟﺎﻜﻴﺳﺭﻭﺲﻳﺯﺎﻳﺮﻴﺘﻴﭘﻥﺍﻮﻨﻋﺖﺤﺗﻲﻌﻳﺎﺷﻲﺘﺳﻮﭘﺖﻧﻮﻔﻋﺎﻫﺎﻳﺰﺳﻻﺎﻣﺯﺍ ﮓﻧﺭﺮﻴﻴﻐﺗﻱﺎﻫﻪﻜﻟﺕﺭﻮﺻﻪﺑﻪﻛﺖﺳﺍ(Pityriasis Versicolor)
ﻭ ﺎﻫﻪﻧﺎﺷ ،ﺖﺸﭘ ،ﻪﻨﻴﺳ ﻲﺣﺍﻮﻧ ﺭﺩ ﺮﺘﺸﻴﺑ ،گﺭﺰﺑ ﻭ ﻚﭼﻮﻛ ﻪﺘﻓﺎﻳ ﻲﻳﺎﻳﺰﺳﻻﺎﻣ ﺖﻴﻟﻮﻜﻴﻟﻮﻓ ﻞﻣﺍﻮﻋ ﺯﺍ ﺎﻫﺎﻳﺰﺳﻻﺎﻣ .ﺩﻮﺷﻲﻣ ﺮﻫﺎﻇ ﻥﺩﺮﮔ
ﻱﺎﻫﻱﺭﺎﻤﻴﺑﺩﺎﺠﻳﺍﺭﺩﺮﻫﺎﻇﻪﺑﻪﻛﻩﺩﻮﺑ (MalasseziaFolliculitis)
،(Seborrheic Dermatitis)ﻱﺍﻩﺭﻮﺷﺖﻴﺗﺎﻣﺭﺩﻞﺜﻣﻱﺮﮕﻳﺩﻲﺘﺳﻮﭘ
(Pseoriasis)ﺲﻳﺯﺎﻳﺭﻮﺴﭘﻭ(Atopic Dermatitis)ﻚﻴﭘﻮﺗﺁﺖﻴﺗﺎﻣﺭﺩ
ﺰﻳﻮﺠﺗﺯﺍﻞﺻﺎﺣﻱﺩﻮﺒﻬﺑﻦﻴﺑﺎﻫﻱﺭﺎﻤﻴﺑﻦﻳﺍﺭﺩ .(2-4) ﺪﻧﺭﺍﺩﺶﻘﻧﺰﻴﻧ
ﻭ ﻮﺳ ﻚﻳﺯﺍ ﺎﻫﺎﻳﺰﺳﻻﺎﻣﺖﻴﻌﻤﺟ ﺶﻫﺎﻛ ﻭ ﻲﭼﺭﺎﻗ ﺪﺿ ﻱﺎﻫﻭﺭﺍﺩ
ﻲﻤﻴﻘﺘﺴﻣ ﻁﺎﺒﺗﺭﺍ ﺮﮕﻳﺩ ﻱﻮﺳ ﺯﺍ ﺖﻴﻌﻤﺟ ﺶﻳﺍﺰﻓﺍﺎﺑ ﻱﺭﺎﻤﻴﺑ ﺖﺸﮔﺮﺑ ﺖﻠﻋﻥﺍﻮﻨﻋ ﻪﺑﺯﻮﻨﻫﺎﻫﺎﻳﺰﺳﻻﺎﻣﺩﻮﺟﻭﻦﻳﺍﺎﺑ .(5) ﺖﺳﺍﻩﺪﺷﻩﺪﻫﺎﺸﻣ
ﻱﺩﺭﺍﻮﻣ،ﺮﻴﺧﺍﺵﺭﺍﺰﮔﺱﺎﺳﺍﺮﺑ .ﺪﻧﺍﻩﺪﺸﻧﻪﺘﻓﺮﻳﺬﭘﺎﻫﻱﺭﺎﻤﻴﺑﻦﻳﺍﻲﻌﻄﻗ
ﻩﮋﻳﻭﻪﺑﺖﻴﻧﻮﻴﺗﺮﭘﻭﻲﻤﺳﻲﺘﭙﺳﻞﺜﻣﻙﺎﻧﺮﻄﺧﻚﻴﻤﺘﺴﻴﺳﻱﺎﻫﺖﻧﻮﻔﻋﺯﺍ ﺪﻳﺪﺷﺕﻼﻜﺸﻣﺭﺎﭼﺩﺍﺭﻲﻋﺎﻓﺩﻢﺘﺴﻴﺳﻪﻛﻦﻴﻐﻟﺎﺑﻭﺱﺭﺎﻧﻥﺍﺩﺍﺯﻮﻧﺭﺩ
.(7 ،6) ﺖﺳﺍﻩﺩﺮﻛ
ﻭ (Basidiomycota) ﺎﺗﻮﻜﻴﻣﻮﻳﺪﻳﺯﺎﺑ ﺔﺧﺎﺷ ﺭﺩ ﺎﻳﺰﺳﻻﺎﻣ ﺲﻨﺟ
.ﺩﺮﻴﮔﻲﻣ ﺭﺍﺮﻗ (Cryptococcaceae) ﻪﺳﺎﻛﻮﻛﻮﺘﭙﻳﺮﻛ ﻩﺩﺍﻮﻧﺎﺧ ﺭﺩ
ﺲﻳﺯﺎﻳﺮﻴﺘﻴﭘﻱﺭﺎﻤﻴﺑﺖﻠﻋﻥﺍﻮﻨﻋﻪﺑ 1846 ﻝﺎﺳﺭﺩﺭﺎﺑﻦﻴﻟﻭﺍﺲﻨﺟﻦﻳﺍ
ﻭ ﺪﺷﻱﺭﺍﺬﮔﻡﺎﻧ Pityrosporum ﻥﺍﻮﻨﻋ ﺖﺤﺗ ﻭ ﻒﺸﻛ ﺮﻟﺎﻜﻴﺳﺭﻭ
ﻦﻳﺍ ﺭﺩP. pachydermatis ﻭ P. ovale ،P.orbicularﻱﺎﻫﻪﻧﻮﮔ
ﻩﺩﺍﺩﻡﺎﻧﺮﻴﻴﻐﺗﺎﻳﺰﺳﻻﺎﻣﻪﺑﻡﻭﺭﻮﭙﺳﻭﺮﻴﺘﻴﭘﺲﭙﺳ .ﺪﻳﺩﺮﮔﻒﻴﺻﻮﺗﺲﻨﺟ
ﻪﻓﺎﺿﺍM. sympodialisﻥﺍﻮﻨﻋﺖﺤﺗﻲﻣﺭﺎﻬﭼﻪﻧﻮﮔ1990 ﻝﺎﺳﺭﺩﻭ .(8) ﺪﻳﺩﺮﮔ
ﺭﺩ ﻩﮋﻳﻭ ﻪﺑ ﻲﻟﻮﻜﻟﻮﻣ ﻱﺎﻫﻱژﻮﻟﻮﻨﻜﺗ ﻪﺑ ﺮﺸﺑ ﻲﺑﺎﻳﺖﺳﺩﻝﺎﺒﻧﺩ ﻪﺑ
ﻭ DNAﺭﺩG+Cﺭﻻﻮﻣﺪﺻﺭﺩ ﻪﻠﻤﺟﺯﺍﻲﻜﻴﺘﻧژﺕﺎﻌﻟﺎﻄﻣﺱﺎﺳﺍﺮﺑ
ﻱژﻮﻟﻮﻳﺰﻴﻓﻭﻱژﻮﻟﻮﻓﺭﻮﻣﺭﺎﻨﻛﺭﺩﻲﻣﻭﺯﻮﺒﻳﺭﻱRNAﻱﺎﻫﻲﻟﺍﻮﺗﺰﻴﻟﺎﻧﺁ M. furfur ،M. pachydermatis ﻞﻣﺎﺷﺎﻳﺰﺳﻻﺎﻣﻪﻧﻮﮔ 7 ،ﺎﻫچﺭﺎﻗ
،M. sympodialis،M. globosa ،M. slooffiae ،M. restricta
ﺱﺎﺳﺍﺮﺑﺮﻴﺧﺍ ﻱﺎﻫﻝﺎﺳ ﺭﺩ .ﺪﺷﻪﺘﻓﺮﻳﺬﭘﻭ ﻒﻴﺻﻮﺗM. obtusa ﻭ
،(10) M. nana ﻥﺍﻮﻨﻋ ﺖﺤﺗ ﺪﻳﺪﺟ ﻪﻧﻮﮔ 4 ﻲﻟﻮﻜﻟﻮﻣ ﻱﺎﻫﻩﺩﺍﺩ (13) M. yamatoensis،(12) M. japonica،(11) M. dermatis
ﻭﻲﻳﺎﺳﺎﻨﺷﺎﻳﺰﺳﻻﺎﻣﻪﻧﻮﮔ 11 ﻞﻛﺭﺩﻪﻛﺪﺷﻪﻓﺎﺿﺍﺎﻫﺎﻳﺰﺳﻻﺎﻣﻊﻤﺟﻪﺑ .ﺖﺳﺍﻩﺪﺷﻪﺘﻓﺮﻳﺬﭘ
26Sﻪﻴﺣﺎﻧ ﻚﻴﺌﻠﻛﻮﻧﻱﺎﻫﺪﻴﺳﺍﻲﻟﺍﻮﺗ ﺱﺎﺳﺍﺮﺑﺮﺿﺎﺣ ﺵﺭﺍﺰﮔ
ﻲﻟﺍﻮﺗﺰﻴﻧﻭ (9-13) ﺖﺳﺍﺪﻳﺪﺟﻱﺎﻫﻪﻧﻮﮔﻒﻴﺻﻮﺗﻱﺎﻨﺒﻣﻪﻛrDNA
ﺮﻛﺭﺎﻣﺰﻴﻧﻥﺁﻪﻛInternal Transcribed Spacer 1 (ITS1)1ﻪﻴﺣﺎﻧ
ﺍﺭ ﺲﻨﺟ ﻦﻳﺍ ﺯﺍ ﻱﺪﻳﺪﺟ ﻪﻟﻭﺰﻳﺍ ،ﺖﺳﺎﻫﻪﻧﻮﮔ ﻕﺍﺮﺘﻓﺍ ﻱﺍﺮﺑ ﻲﻤﻬﻣ
ﻪﻛﺪﻳﺪﺟﻪﻟﻭﺰﻳﺍﻦﻳﺍﺺﺧﺎﺷﻰﻟﻮﻜﻟﻮﻣﻱﺎﻫﺕﻭﺎﻔﺗ .ﺪﻳﺎﻤﻧﻰﻣﻰﻓﺮﻌﻣ
ﻭﻞﻴﻤﻜﺗﺯﺍﺲﭘﺩﻮﺷﻲﻣﻱﺯﺎﺳﺍﺪﺟﻥﺍﺮﻳﺍﺯﺍﻥﺎﻬﺟﺭﺩﺭﺎﺑﻦﻴﻟﻭﺍﻱﺍﺮﺑ ﻱﺍﻪﻧﻮﮔﻑﺮﻌﻣﻚﻳژﻮﻟﻮﻳﺰﻴﻓﻭﻚﻳژﻮﻟﻮﻓﺭﻮﻣﺕﺎﻋﻼﻃﺍﺮﻳﺎﺳﻒﻴﺻﻮﺗ
.ﺩﻮﺷﻲﻣﺪﻳﺪﺟ
ﺩﺭﻮﻣ
ﺵﺭﺍﺰﮔ
ﺮﻤﺨﻣﻱﺯﺎﺳﺍﺪﺟ
ﻡﺎﺠﻧﺍ ﻰﻠﻫﺍﺕﺎﻧﺍﻮﻴﺣ ﻲﻳﺎﻳﺰﺳﻻﺎﻣ ﺭﻮﻠﻓ ﺮﺑﻪﻛ ﻱﺍﻪﻌﻟﺎﻄﻣ ﻲﻃ ﺭﺩ
MLANﻪﺘﻓﺎﻳﺮﻴﻴﻐﺗ ﻂﻴﺤﻣﻱﻭﺭ ﺏﺁﻮﺳ ﻭ ﻪﺘﺳﻮﭘ ﻱﺎﻫﻪﻧﻮﻤﻧ،ﺪﺷﻲﻣ
،(ﺪﺻﺭﺩ1) ﻥﻮﺘﭙﭘ ﻞﻣﺎﺷ (14) (Leeming and Notman)
ﻱﻭﺎﮔ ﻱﻭﺍﺮﻔﺻ ،(ﺪﺻﺭﺩ0/2) ﺮﻤﺨﻣ ﻩﺭﺎﺼﻋ ،(ﺪﺻﺭﺩ1) ﺰﻛﻮﻠﮔ
ﺕﺍﺭﺎﺋﺎﺘﺳﺍﻮﻧﻮﻣ ﻝﻭﺮﺴﻴﻠﮔ ،(ﺪﺻﺭﺩ1) ﻝﻭﺮﺴﻴﻠﮔ ،(ﺪﺻﺭﺩ0/8) (ﺪﺻﺭﺩ2) ﻥﻮﺘﻳﺯ ﻦﻏﻭﺭ ،(ﺪﺻﺭﺩ0/5) 60 ﻦﻴﻳﻮﺗ ،(ﺪﺻﺭﺩ0/05)
ﻲﻜﻳ ﻯﺎﻫﻪﻧﻮﻤﻧ ﻦﻴﺑ ﺯﺍ .ﺪﺷ ﻩﺩﺍﺩ ﺖﺸﻛ (ﺪﺻﺭﺩ1/5) ﺭﺎﮔﺁ ﻭ
ﻱﻮﮕﻟﺍ ﻪﻛ ﺪﺷ ﺍﺪﺟ ﻱﺮﻤﺨﻣ ،ﻪﻌﻟﺎﻄﻣ ﺩﺭﻮﻣ ﻯﺎﻫﺮﺘﺴﻣﺎﻫ ﺯﺍ
ﻥﺁ(Restriction Fragment Length Polymorphism)ﻭRFLP
ﻭﺖﺷﺍﺩﺕﻭﺎﻔﺗ،(15) ﺪﻧﺍﻩﺪﺷﻲﻳﺎﺳﺎﻨﺷﻝﺎﺣﻪﺑﺎﺗﻪﻛﻲﻳﺎﻫﺎﻳﺰﺳﻻﺎﻣﺎﺑ .ﺪﻳﺩﺮﮔﻥﺁﻱﻭﺭﺮﺑﺮﺘﺸﻴﺑﺕﺎﻌﻟﺎﻄﻣﺐﺟﻮﻣﺎﻫﺕﻭﺎﻔﺗﻦﻴﻤﻫ
ﻱﺮﻤﺨﻣﻲﻧﻮﻠﻛﺯﺍﻲﻣﻮﻧژﻱDNAﺝﺍﺮﺨﺘﺳﺍ
پﻮﻟ ﻚﻳ (16) ﺪﺷ ﻩﺩﺍﺩ ﺢﻴﺿﻮﺗ ﻪﻛ ﻲﺷﻭﺭ ﻖﺑﺎﻄﻣ
ﻲﻧﻮﻠﻛﺯﺍ (ﺐﻌﻜﻣ ﺮﺘﻣﻲﻠﻴﻣ 5 ﺩﻭﺪﺣ) ﻱژﻮﻟﻮﻳﺮﺘﻛﺎﺑ
ﻩﺩﺮﻛ ﻞﻘﺘﻨﻣ ﻑﺭﺪﻨﭘﺍ ﻱﺮﺘﻴﻟﻲﻠﻴﻣ 1/5 ﺏﻮﻴﺗ ﻪﺑ ﻭ ﻪﺘﺷﺍﺩﺮﺑ
ﺲﻳﺮﺗ ﺭﻻﻮﻣﻲﻠﻴﻣ 100 ﻞﻣﺎﺷ ﺰﻴﻟ ﺮﻓﺎﺑ ﺮﺘﻴﻟﻭﺮﻜﻴﻣ 300 ﻭ ﺪﺻﺭﺩ2 ،SDS ﺪﺻﺭﺩ1 ،NaClﺭﻻﻮﻣﻲﻠﻴﻣ 100 ،EDTAﻭ(pH 8)
ﺩﻭﺪﺣﻭ (1:1) ﻡﺮﻓﻭﺮﻠﻛﻞﻨﻓﻁﻮﻠﺨﻣﺮﺘﻴﻟﻭﺮﻜﻴﻣ 300 ،Triton X-100
ﺮﺘﻣﻲﻠﻴﻣ 1 ﺮﻄﻗﻪﺑ(glass-beads)ﻱﺍﻪﺸﻴﺷﻱﺎﻫﻝﺮﭘﺮﺘﻴﻟﻭﺮﻜﻴﻣ 200
ﻩﺎﮔﻥﺁ .ﺪﺷﻩﺩﺍﺩﻥﺎﻜﺗﺕﺪﺷﻪﺑﺖﺳﺩﺎﺑﻪﻘﻴﻗﺩ 3 ﺕﺪﻣﻭﺪﻳﺩﺮﮔﻪﻓﺎﺿﺍ
ﺪﻳﺪﺟ ﺏﻮﻴﺗ ﻪﺑ ﻲﻳﻭﺭ ﻊﻳﺎﻣ ﻭ ژﻮﻴﻔﻳﺮﺘﻧﺎﺳ 5000 ﺭﻭﺩ ﺭﺩ ﻪﻘﻴﻗﺩ 5
ﺩﺪﺠﻣژﻮﻴﻔﻳﺮﺘﻧﺎﺳﺯﺍﺲﭘﺪﺷﻪﻓﺎﺿﺍﻡﺮﻓﻭﺮﻠﻛﺮﺘﻴﻟﻭﺮﻜﻴﻣ 400 ﻭﻞﻘﺘﻨﻣ
ﺩﺮﺳﻝﻮﻧﺎﭘﻭﺮﭘﻭﺰﻳﺍﻢﺠﺣ 1 ﺭﺍﺪﻘﻣﺪﻳﺪﺟﺏﻮﻴﺗﻪﺑﻲﻳﻭﺭﻊﻳﺎﻣﻝﺎﻘﺘﻧﺍﻭ
ﺯﺍﺲﭘﻭﻩﺩﺮﻛﻪﻓﺎﺿﺍ (pH= 5/2) ﺭﻻﻮﻣ 3 ﻢﻳﺪﺳﺕﺎﺘﺳﺍﻢﺠﺣ 0/1 ﻭ
ﻪﻘﻴﻗﺩ 15 ﺕﺪﻣ ،ﺩﺍﺮﮔﻲﺘﻧﺎﺳ ﻪﺟﺭﺩ -20 ﺭﺩ ﻱﺭﺍﺪﻬﮕﻧ ﻪﻘﻴﻗﺩ 10
ﻱﻮﺷﻭﺖﺴﺷﺯﺍ ﺲﭘﺖﻳﺎﻬﻧ ﺭﺩﻭ ﻩﺪﺷژﻮﻴﻔﻳﺮﺘﻧﺎﺳ 10000 ﺭﻭﺩﺭﺩ
ﺪﻳﺩﺮﮔﻞﺣﺏﺁﺮﺘﻴﻟﻭﺮﻜﻴﻣ 50 ﺭﺩﻪﺟﺭﺩ 70 ﻞﻜﻟﺍﺎﺑﻪﻠﺻﺎﺣﺏﻮﺳﺭ .ﺪﺷﻱﺭﺍﺪﻬﮕﻧﺩﺍﺮﮔﻲﺘﻧﺎﺳﻪﺟﺭﺩ -20 ﺭﺰﻳﺮﻓﺭﺩﻡﺯﻻﻥﺎﻣﺯﺎﺗﻭ (PCR)ﺯﺍﺮﻤﻴﻠﭘﻱﺍﻩﺮﻴﺠﻧﺯﺶﻨﻛﺍﻭ
ﺮﻤﻳﺍﺮﭘ ﺖﻔﺟ ﻩﺪﺷ ﻡﺎﺠﻧﺍ PCR ﺮﻤﻳﺍﺮﭘ ﺖﻔﺟ ﻭﺩ ﺯﺍ ﻩﺩﺎﻔﺘﺳﺍ ﺎﺑ
ﻭ (Mal R: 5'-ATTACGCCAGCATCCTAAG-3'3) ﻝﻭﺍ
Mal F: 5'-TAACAAGGATCCCCCTAGTA-3'3)
ﺖﻬﺟ ،(15) ﺩﻮﺑ ﻩﺪﺷ ﻲﺣﺍﺮﻃ ﻒﻟﻮﻣ ﻲﻠﺒﻗ ﺕﺎﻌﻟﺎﻄﻣ ﺭﺩ ﻪﻛ
ﻪﺑ ﻁﻮﺑﺮﻣ 26S ﻪﻴﺣﺎﻧﺭﺩ ﺩﻮﺟﻮﻣ D1-D2 ﻪﻌﻄﻗ ﺖﻳﻮﻘﺗ
ﺮﻤﻳﺍﺮﭘ ﺖﻔﺟﻭ ﺖﻓﺮﮔ ﺭﺍﺮﻗ ﻩﺩﺎﻔﺘﺳﺍ ﺩﺭﻮﻣ ﻲﻣﻭﺯﻮﺒﻳﺭ ﻱDNA
ﻭ 18SF1: 5'-AGGTTTCCGTAGGTGAACCT-3') ﻡﻭﺩ
ﺯﺍ ﻪﻛ (58SR1: 5'-TTCGCTGCGTTCTTCATCGA-3')
ﻪﻴﺣﺎﻧ ﺖﻳﻮﻘﺗ ﺖﻬﺟ ﺪﺷﻪﺘﻓﺮﮔ (17) ﻥﺍﺭﺎﻜﻤﻫ ﻭ ﺍﺭﻮﻤﻴﻛﺎﻣ ﺵﺭﺍﺰﮔ
ﻪﺑﻲﻣﻭﺯﻮﺒﻳﺭﻱDNAﻪﺑﻁﻮﺑﺮﻣ18Sﻭ 5.8SﺕﺎﻌﻄﻗﻦﻴﺑﺝﺭﺪﻨﻣITS1
ﻩﺪﺷﺝﺍﺮﺨﺘﺳﺍﻱDNAﺯﺍﺮﺘﻴﻟﻭﺮﻜﻴﻣﻚﻳPCRﻡﺎﺠﻧﺍﻱﺍﺮﺑ .ﺖﻓﺭﺭﺎﻛ
ﻡﺍﺪﻛﺮﻫﺯﺍ (ﺭﻻﻮﻣﻭﺮﻜﻴﻣ 0/5) ﺮﺘﻴﻟﻭﺮﻜﻴﻣ 1 ،PCRﺮﻓﺎﺑﺮﺘﻴﻟﻭﺮﻜﻴﻣ 5 ﻪﺑ
-Taq DNA polymerﻢﻳﺰﻧﺁ (ﺪﺣﺍﻭ 2/5) ﺮﺘﻴﻟﻭﺮﻜﻴﻣ 0/5 ،ﺎﻫﺮﻤﻳﺍﺮﭘﺯﺍ
Tﻱﺎﻫﺪﻴﺗﻮﺌﻠﻛﻮﻧ) dNTPs (ﺭﻻﻮﻣﻭﺮﻜﻴﻣ200) ﺮﺘﻴﻟﻭﺮﻜﻴﻣ 0/5 ﻭase 50ﻪﺑﻞﻳﺮﺘﺳﺍﺮﻄﻘﻣﺏﺁﺎﺑﻲﻳﺎﻬﻧﻢﺠﺣﻭﺪﻳﺩﺮﮔﻪﻓﺎﺿﺍ (Aﻭ ،C ،G
95 ﻞﻣﺎﺷPCRﺶﻨﻛﺍﻭﻭﺩﺮﻫﺭﺩPCRﻪﻣﺎﻧﺮﺑ .ﺪﺷﻩﺪﻧﺎﺳﺭﺮﺘﻴﻟﻭﺮﻜﻴﻣ
45 ﺕﺪﻣﻪﺑﻪﺟﺭﺩ 94 ،ﻞﻜﻴﺳﻚﻳﻪﻘﻴﻗﺩ 5 ﺕﺪﻣﻪﺑﺩﺍﺮﮔﻲﺘﻧﺎﺳﻪﺟﺭﺩ
30 ،ﻪﻘﻴﻗﺩ 1 ﺕﺪﻣﻪﺑﻪﺟﺭﺩ 72 ﻭﻪﻘﻴﻗﺩﻚﻳﺕﺪﻣﻪﺑﻪﺟﺭﺩ 56 ،ﻪﻴﻧﺎﺛ
.ﺩﻮﺑﻞﻜﻴﺳﻚﻳﻪﻘﻴﻗﺩ 7 ﺕﺪﻣﻪﺑﻪﺟﺭﺩ 72 ﻭﻞﻜﻴﺳ
RFLP
ﻩﺩﺎﻔﺘﺳﺍ ﺎﺑ ،ﻩﺪﺷ ﺍﺪﺟ ﻱﺎﻳﺰﺳﻻﺎﻣ ﻪﻧﻮﮔ ﻪﻴﻟﻭﺍ ﻲﻳﺎﺳﺎﻨﺷ ﺭﻮﻈﻨﻣ ﻪﺑ ﻢﻀﻫ ﻪﺑ ﻡﺍﺪﻗﺍ ،(15) ﺖﺳﺍ ﻩﺪﺷ ﺵﺭﺍﺰﮔ ﻞﺒﻗ ﺯﺍ ﻪﻛ ﻲﻠﻴﻓﻭﺮﭘ ﺯﺍ
ﻪﺑ .ﺪﻳﺩﺮﮔ 26S ﻪﻴﺣﺎﻧ ﻪﺑ ﻁﻮﺑﺮﻣ PCR ﺕﻻﻮﺼﺤﻣ ﻱﺯﺎﺌﻠﻛﻮﻧﻭﺪﻧﺍ
ﻢﻳﺰﻧﺁ ﺮﺘﻴﻟﻭﺮﻜﻴﻣ 1 ﺎﺑ PCRﻝﻮﺼﺤﻣ ﺯﺍﺮﺘﻴﻟﻭﺮﻜﻴﻣ 10 ﻪﺻﻼﺧﺭﻮﻃ
ﻲﻳﺎﻬﻧﻢﺠﺣﺎﺗﺮﻄﻘﻣﺏﺁﻭﻁﻮﻠﺨﻣﻪﻃﻮﺑﺮﻣﺮﻓﺎﺑﺮﺘﻴﻟﻭﺮﻜﻴﻣ 2/5 ﻭCfoI
ﻪﺟﺭﺩ 37 ﺕﺭﺍﺮﺣﺭﺩ ﺖﻋﺎﺳ 2ﺕﺪﻣﻭ ﻩﺪﻳﺩﺮﮔﻪﻓﺎﺿﺍ ﺮﺘﻴﻟﻭﺮﻜﻴﻣ25 .ﺪﺷﻪﺑﻮﻜﻧﺍﺩﺍﺮﮔﻲﺘﻧﺎﺳ
ﺯﺭﻮﻓﻭﺮﺘﻜﻟﺍ
ﻚﻳ ﺕﺪﻣ ﻪﺑ ﺪﺻﺭﺩ 1/5 ﺯﺭﺎﮔﺁ ﻝژ ﻱﻭﺭPCR ﺕﻻﻮﺼﺤﻣ
ﺖﻋﺎﺳ 2 ﺕﺪﻣ ﻪﺑ ﺯﺭﺎﮔﺁ ﻝژ ﻱﻭﺭ RFLP ﺕﻻﻮﺼﺤﻣ ﻭ ﺖﻋﺎﺳ
ﺭﻻﻮﻣ 0/09 ،ﺲﻳﺮﺗ ﺭﻻﻮﻣ 0/09) TBE ﺮﻓﺎﺑ .ﺪﻳﺩﺮﮔ ﺯﺭﻮﻓﻭﺮﺘﻜﻟﺍ
ﺖﺧﺎﺳ ﺖﻬﺟ (pH= 3/8 ،EDTA ﺭﻻﻮﻣﻲﻠﻴﻣ 2 ﻭ ﻚﻳﺭﻮﺑ ﺪﻴﺳﺍ
ﻥﺎﻳﺮﺟ ﻚﻤﻛ ﻪﺑ ﺎﻫﻪﻧﻮﻤﻧ .ﺖﻓﺭ ﺭﺎﻛ ﻪﺑ ﻚﻧﺎﺗ ﻥﺩﺮﻛ ﺮﭘ ﻭ ﻝژ
ﻝﻮﻃ ﺮﺘﻣﻲﺘﻧﺎﺳ ﺮﻫﻱﺍﺯﺍ ﻪﺑ ﺖﻟﻭ 5-10 ژﺎﺘﻟﻭ ﺎﺑ ﻢﻴﻘﺘﺴﻣ ﻪﺘﻴﺴﻳﺮﺘﻜﻟﺍ
ﻪﺑﺪﻴﻣﻭﺮﺑﻡﻮﻳﺪﻴﺗﺍﮓﻧﺭﺎﺑ ﻪﻘﻴﻗﺩ 20 ﺕﺪﻣﻭﺪﻳﺩﺮﮔﺯﺭﻮﻓﻭﺮﺘﻜﻟﺍﻝژ 312 ﺝﻮﻣﻝﻮﻃﺎﺑﻭﻱﺰﻴﻣﺁﮓﻧﺭﺮﺘﻴﻟﻲﻠﻴﻣﺭﺩﻡﺮﮔﻭﺮﻜﻴﻣ 0/5 ﺖﻈﻠﻏ
ﻝﺎﺘﻴﺠﻳﺩﻖﻳﺮﻃ ﻪﺑﻭﻩﺪﻫﺎﺸﻣﺭﻮﺗﺎﻨﻴﻣﻮﻟﺍﺲﻧﺍﺮﺗﺯﺍﻩﺩﺎﻔﺘﺳﺍﺎﺑﻭﺮﺘﻣﻮﻧﺎﻧ
.ﺪﺷﻱﺭﺍﺩﺮﺑﺲﻜﻋ
(Sequencing) ﻲﻟﺍﻮﺗﻦﻴﻴﻌﺗ
ﻱﺍﺮﺑﻭﺖﺸﮔﺮﺑﻭﺖﻓﺭﺮﻤﻳﺍﺮﭘﻭﺩﺯﺍ26Sﻪﻴﺣﺎﻧﻲﻟﺍﻮﺗﻦﻴﻴﻌﺗﻱﺍﺮﺑ
ﺎﺑﺎﻫﻪﻧﻮﻤﻧ .ﺪﺷﻩﺩﺎﻔﺘﺳﺍﺖﻓﺭﺮﻤﻳﺍﺮﭘﺯﺍﻂﻘﻓ ITS1ﻪﻴﺣﺎﻧﻲﻟﺍﻮﺗﻦﻴﻴﻌﺗ .ﺪﻳﺩﺮﮔﻲﻟﺍﻮﺗﻦﻴﻴﻌﺗ Prism-ABIﻢﺘﺴﻴﺳﺯﺍﻩﺩﺎﻔﺘﺳﺍ
ﺎﻫﻩﺩﺍﺩﻱﺮﺗﻮﻴﭙﻣﺎﻛﺰﻴﻟﺎﻧﺁ
ﺎﺑ Blast ﺭﺍﺰﻓﺍﻡﺮﻧ ﺯﺍ ﻩﺩﺎﻔﺘﺳﺍ ﺎﺑ ﻲﻟﺍﻮﺗ ﻦﻴﻴﻌﺗ ﻱﺎﻫﻩﺩﺍﺩ
ﺮﻳﺎﺳ ﻭ ﺎﻫﺎﻳﺰﺳﻻﺎﻣ ﻪﺑ ﻁﻮﺑﺮﻣ ﻥژ ﻚﻧﺎﺑ ﺭﺩ ﺝﺭﺪﻨﻣ ﻱﺎﻫﻩﺩﺍﺩﺮﻳﺎﺳ ﻁﻮﺑﺮﻣ ﻱﺎﻫﺲﻧﺎﻜﺳ ﻪﺴﻳﺎﻘﻣ ﻱﺍﺮﺑ .ﺪﺷ ﻪﺴﻳﺎﻘﻣ ﺎﻫﻢﺴﻴﻧﺎﮔﺭﺍ ﻭﺮﻜﻴﻣ
ClastalW ﻭ DNASIS ﻱﺎﻫﺭﺍﺰﻓﺍﻡﺮﻧ ﺯﺍ ﻒﻠﺘﺨﻣﻱﺎﻫﺎﻳﺰﺳﻻﺎﻣﻪﺑ
.ﺪﺷ ﻩﺩﺎﻔﺘﺳﺍﺰﻴﻧ
ﺎﻫ
ﻪﺘﻓﺎﻳ
(1 ﻞﻜﺷ) MLANﺭﺍﺩﻲﺑﺮﭼﻂﻴﺤﻣﻱﻭﺭﻪﺘﻓﺎﻳﺪﺷﺭﻱﺎﻫﻲﻧﻮﻠﻛ
(2 ﻞﻜﺷ) ﻲﻧﻮﻠﻛﻱﺮﻤﺨﻣﻱﺎﻫﻝﻮﻠﺳﻲﭘﻮﻜﺳﻭﺮﻜﻴﻣﻱژﻮﻟﻮﻓﺭﻮﻣﺰﻴﻧﻭ
.ﺩﻮﺑﺎﻳﺰﺳﻻﺎﻣﺲﻨﺟﺯﺍﻲﻳﺎﻫﺮﻤﺨﻣﺩﻮﺟﻭﺯﺍﺖﻳﺎﻜﺣ
ﺮﻴﻴﻐﺗﻂﻴﺤﻣﻯﻭﺭﻪﺘﻓﺎﻳﺪﺷﺭﺎﻳﺰﺳﻻﺎﻣﺪﻳﺪﺟﻪﻟﻭﺰﻳﺍﻯﺎﻫﻰﻧﻮﻠﻛ :1 ﻞﻜﺷ Leeming and Notman ﻪﺘﻓﺎﻳ
ﺪﻳﺪﺟ ﻪﻟﻭﺰﻳﺍ ﻱﺮﻤﺨﻣ ﻱﺎﻫﻝﻮﻠﺳ ﻲﭘﻮﻜﺳﻭﺮﻜﻴﻣ ﻱژﻮﻟﻮﻓﺭﻮﻣ :2 ﻞﻜﺷ ﺎﻳﺰﺳﻻﺎﻣ
ﻒﻳﺩﺭ) 26SrDNA D1/D2ﻰﺣﺍﻮﻧPCRﺕﻻﻮﺼﺤﻣﺯﺭﻮﻓﻭﺮﺘﻜﻟﺍ :3 ﻞﻜﺷ ﻲﻟﻮﻜﻟﻮﻣﺮﻛﺭﺎﻣ :Mﻒﻳﺩﺭ .ﺪﻳﺪﺟﻪﻟﻭﺰﻳﺍﻪﺑﻁﻮﺑﺮﻣ (2 ﻒﻳﺩﺭ) ITS1ﻭ (1
500 bp
300 bp
.ﻱﺯﺎﺑﺖﻔﺟ 100
ﺯﺍﺲﭘﺪﺻﺭﺩ 2 ﺯﺭﺎﮔﺁﻝژﻯﻭﺭPCRﺕﻻﻮﺼﺤﻣﺯﺭﻮﻓﻭﺮﺘﻜﻟﺍ :4 ﻞﻜﺷ
،ﺍﺯﻮﺑﻮﻠﮔﺎﻳﺰﺳﻻﺎﻣ :ﺐﻴﺗﺮﺗﻪﺑ 12 ﺎﺗ 1 ﻯﺎﻫﻚﻫﺎﭼ .CfoIﻢﻳﺰﻧﺁﺎﺑﺵﺮﺑ .ﻡ،ﻪﻴﻓﻮﻠﺳﺍ .ﻡ،ﺎﺳﻮﺘﺑﺍ .ﻡ ،ﺲﻴﻟﺎﻳﺩﻮﭙﻤﺳ .ﻡ ،ﺭﻮﻓﺭﻮﻓ .ﻡ ،ﺎﺘﻜﻳﺮﺘﺳﺭ .ﻡ ،ﺲﻴﺴﻨﺋﻮﺗﺎﻣﺎﻳ .ﻡ ،ﺎﻧﺎﻧ .ﻡ ،ﺲﻴﺗﺎﻣﺭﺩ ﻰﻛﺎﭘ .ﻡ ،ﺲﻴﺗﺎﻣﺭﺩ .ﻡ ،ﺎﻜﻴﻧﻮﭘﺍژ
.ﻯﺯﺎﺑﺖﻔﺟ 100 ﻰﻟﻮﻜﻟﻮﻣﺮﻛﺭﺎﻣMﻚﻫﺎﭼﻭﻩﺪﺷﻒﺸﻛﻩﺯﺎﺗﻯﺎﻳﺰﺳﻻﺎﻣ
ﻪﺑ (12 ﻒﻳﺩﺭ) ﺪﻳﺪﺟﻯﺎﻳﺰﺳﻻﺎﻣﻚﻴﺗﺭﻮﻓﻭﺮﺘﻜﻟﺍﻯﻮﮕﻟﺍﻪﻛﺩﻮﺷﻰﻣﻪﻈﺣﻼﻣ .ﺩﺭﺍﺩﺕﻭﺎﻔﺗﻪﻴﻘﺑﺎﺑﺡﻮﺿﻭ
ﺮﺘﺸﻴﺑﺵﺎﻜﻨﻛﺚﻋﺎﺑﻭﺩﻮﺑﻪﻟﻭﺰﻳﺍﻥﺩﻮﺑﺪﻳﺪﺟﻞﻴﻟﺩﻦﻴﻟﻭﺍﻉﻮﺿﻮﻣﻦﻳﺍ
:ﺩﻮﺷﻰﻣﻩﺪﻳﺩﺮﻳﺯﺭﺩﺭﻮﺑﺰﻣﻰﻟﺍﻮﺗ .ﺪﻳﺩﺮﮔﻞﻣﺎﻛﻲﻟﺍﻮﺗﻦﻴﻴﻌﺗﺖﻳﺎﻬﻧﺭﺩﻭ
A N A N T T G N A G C T G G C G C C T C C G G C G T C C G C G T G TA AT C T C G A G A C G T G T T T T C C G T G C G G C A C T G T- GGACAAGTTCTCTGGAAAGGGACATCGTAGAGGGT- GAAAATCCCGTACTTGCCACGGCTGAACCGTGCTTTGCGA- TACACGCTCTAAGAGTCGAGTTGTTTGGGATTGCAGCT- CAAAGTGGGTGGTAGACTCCATCTAAAGCTAAATATCG- GGGAGAGACCGATAGCGAACAAGTACCGTGAGGGAAA- GATGAAAAGCACTTTGGAAAGAGAGTTAAAAGTACGT- GAAATTGTCGAAAGGGAAGCGCTTGGAGTTAGCCAT- GCCGCTCGAGATTCAGCCTTACGGTGTACTTCTCGGGTAG- CAAGTCAGCATTGGTTTGTGCCGCTGGAGAAGGGCGT-G CAAGTCAGCATTGGTTTGTGCCGCTGGAGAAGGGCGT-G CAAGTCAGCATTGGTTTGTGCCGCTGGAGAAGGGCGT-G CAAGTCAGCATTGGTTTGTGCCGCTGGAGAAGGGCGT-G A AT CAAGTCAGCATTGGTTTGTGCCGCTGGAGAAGGGCGT-G T CAAGTCAGCATTGGTTTGTGCCGCTGGAGAAGGGCGT-G CAAGTCAGCATTGGTTTGTGCCGCTGGAGAAGGGCGT-G C A C T T C CAAGTCAGCATTGGTTTGTGCCGCTGGAGAAGGGCGT-G CAAGTCAGCATTGGTTTGTGCCGCTGGAGAAGGGCGT-G T CAAGTCAGCATTGGTTTGTGCCGCTGGAGAAGGGCGT-G T CAAGTCAGCATTGGTTTGTGCCGCTGGAGAAGGGCGT-G T TATA CAAGTCAGCATTGGTTTGTGCCGCTGGAGAAGGGCGT-G C C C C ATG C T ATG ATG ATA C A ATG C ATG ATG C ATG C ATG ATG AT C A A ATG ATG A A C ATG C A ATG - CACGCCTTGTGGCGGGTCTTCGGACACCTTCGTGCTTAG-GANNNNNNNCGTAATA
ﺎﻫﺎﻳﺰﺳﻻﺎﻣﺮﻳﺎﺳﺎﺑﻪﻟﻭﺰﻳﺍﻦﻳﺍﻪﻛﺪﺷﺺﺨﺸﻣﺎﻫﻰﻟﺍﻮﺗﺰﻴﻟﺎﻧﺁﺯﺍﺲﭘ ﻚﻴﻧژﻮﻠﻴﻓ ﺖﺧﺭﺩ 5 ﻞﻜﺷ .ﺩﺭﺍﺩ ﺕﻭﺎﻔﺗ ﻲﻬﺟﻮﺗ ﻞﺑﺎﻗ ﻥﺍﺰﻴﻣ ﻪﺑ
ﺭﺩ 26S ﻪﻴﺣﺎﻧ ﻲﻟﺍﻮﺗ ﻪﺴﻳﺎﻘﻣ ﺯﺍ ﺲﭘ ﻪﻛ ﺍﺭ (Phylogentic tree)
،ﺖﺳﺍ ﻩﺪﺷ ﻢﺳﺭ DNAsis ﺭﺍﺰﻓﺍﻡﺮﻧﻚﻤﻛ ﻪﺑ ﻒﻠﺘﺨﻣ ﻱﺎﻫﺎﻳﺰﺳﻻﺎﻣ
.ﺪﻫﺩﻰﻣﻥﺎﺸﻧ
ﻪﻌﻟﺎﻄﻣﻦﻳﺍﻲﻃﻩﺪﺷﻒﺸﻛﺪﻳﺪﺟﻱﺎﻳﺰﺳﻻﺎﻣﺖﺧﺭﺩﻦﻳﺍﻖﺑﺎﻄﻣ ﻝﻮﺼﺣﺭﻮﻈﻨﻣﻪﺑ .ﺩﺭﺍﺩﺭﺍﺮﻗﻪﺧﺎﺷﻚﻳﻱﻭﺭM. sloofieaﺭﺎﻨﻛﺭﺩ
ﺎﺑ ITS1ﻪﻴﺣﺎﻧﻩﺪﺷﺍﺪﺟﻪﻟﻭﺰﻳﺍﻥﺩﻮﺑﺪﻳﺪﺟﺎﺑﻪﻄﺑﺍﺭﺭﺩﺮﺘﺸﻴﺑﻥﺎﻨﻴﻤﻃﺍ .ﺖﻓﺮﮔﺕﺭﻮﺻﻲﻟﺍﻮﺗﻦﻴﻴﻌﺗﻭ PCR،ﻪﻃﻮﺑﺮﻣﺮﻤﻳﺍﺮﭘﺖﻔﺟﺯﺍﻩﺩﺎﻔﺘﺳﺍ
280 ﺩﻭﺪﺣ ﻥﺯﻭ ﻪﺑ ﺪﻧﺎﺑ ﻚﻳ PCR ﻝﻮﺼﺤﻣ ﺯﺭﻮﻓﻭﺮﺘﻜﻟﺍ ﺯﺍ ﺲﭘ
ﺝﺭﺩﺮﻳﺯﺭﺩITS1ﻪﻴﺣﺎﻧﻰﻟﺍﻮﺗ .(3 ﻞﻜﺷ) ﺪﻳﺩﺮﮔﻩﺪﻫﺎﺸﻣ ﺯﺎﺑﺖﻔﺟ :ﺩﻮﺷﻰﻣ
G N G C T G N G G N A G AT G T G A G AT G G C G G G C T T T C - GAGCCCGCACGCACTGTCCACCTCACCTCCACACAGATC- CAAACCCCTGTGCACACGCTCTCTCTGGGCGCGCG- GAAATGCCGCAAGCCTGGGGGGAAGAATGAAACACC TA G GAAATGCCGCAAGCCTGGGGGGAAGAATGAAACACC GAAATGCCGCAAGCCTGGGGGGAAGAATGAAACACC A G T T G A AT G A A GAAATGCCGCAAGCCTGGGGGGAAGAATGAAACACC G T TA G T G T G T T G G AT GAAATGCCGCAAGCCTGGGGGGAAGAATGAAACACC G - TAACGACCAACGAAGAAAAACACAACTTTCGACAACG-GATCTCTAGGTTCTCCCATCGATGAAGA
ﺪﻳﺪﺟﻯﺎﻳﺰﺳﻻﺎﻣﻩﺎﮕﻳﺎﺟﻙﺭﺩﻯﺍﺮﺑﺍﺭ ﻚﻴﻧژﻮﻠﻴﻓﺖﺧﺭﺩ 6 ﻞﻜﺷ
ﻥﺎﺸﻧ ،ﻒﻠﺘﺨﻣ ﻱﺎﻫﺎﻳﺰﺳﻻﺎﻣ ﺭﺩ ITS ﻪﻴﺣﺎﻧ ﻲﻟﺍﻮﺗ ﻪﺴﻳﺎﻘﻣ ﻪﺑ ﻪﺟﻮﺗ ﺎﺑ .ﺪﻫﺩﻲﻣ
500 bp
ﺚﺤﺑ
ﺖﺳﻮﭘ ﻲﺑﺮﭼ ﻱﺎﻫﺮﻤﺨﻣ ﻱﺪﻨﺑﻪﻘﺒﻃ ﻙﻼﻣ ﻪﻛ 1995 ﻝﺎﺳ ﺯﺍ
ﻚﻳژﻮﻟﻮﻓﺮﻣﻭﻚﻳژﻮﻟﻮﻳﺰﻴﻓﻱﺎﻫﺺﺧﺎﺷﺯﺍﺎﻳﺰﺳﻻﺎﻣﺲﻨﺟﻪﺑﻖﻠﻌﺘﻣ ﻪﻌﺳﻮﺗﻪﻧﻮﮔ 7 ﻪﺑﺎﻳﺰﺳﻻﺎﻣﻪﻧﻮﮔ 3 ﻭﺩﻮﻤﻧﺮﻴﻴﻐﺗﻲﭙﻴﺗﻮﻧژﻱﺎﻫﺮﻛﺭﺎﻣﻪﺑ 11 ﻊﻤﺟﺭﺩﻭﻩﺪﺷﻪﻓﺎﺿﺍﺎﻬﻧﺁﻪﺑﺪﻳﺪﺟﻪﻧﻮﮔ 4 ﻥﻮﻨﻛﺎﺗ .(9) ﺩﺮﻛﺍﺪﻴﭘ
.(10-13) ﺖﺳﺍﻩﺪﺷﻪﺘﻓﺮﻳﺬﭘﻭﻒﻴﺻﻮﺗﺎﻳﺰﺳﻻﺎﻣﻪﻧﻮﮔ
ﻱﺎﻫﺶﺨﺑ ﻱﺍﺭﺍﺩ (rDNA) ﻲﻣﻭﺯﻮﺒﻳﺭ ﻱDNA ﻲﻧژ ﺲﻜﻠﭙﻤﻛ
ﺖﺳﺍ (Variable) ﺮﻴﻐﺘﻣ ﻭ (Conserved) ﻩﺪﺷﺖﻈﻓﺎﺤﻣ ﻒﻠﺘﺨﻣ
ﻲﻳﺎﺳﺎﻨﺷ ﻱﺍﺮﺑ ﺪﻧﺍﻮﺗﻲﻣ ﻲﻘﻴﻘﺤﺗ ﺎﻳ ﻲﺼﻴﺨﺸﺗ ﻑﺍﺪﻫﺍ ﻪﺑ ﻪﺘﺴﺑ ﻪﻛ ﻪﻧﻮﮔﺮﻳﺯ ﻭ ﺲﻨﺟ ﻕﻮﻓ ﺎﻳ ﻪﻧﻮﮔﻭ ﺲﻨﺟ ﺪﺣ ﺭﺩ ﺎﻫﻢﺴﻴﻧﺎﮔﺭﺍﻭﺮﻜﻴﻣ ﻦﻴﻴﻌﺗ ﻱﺍﺮﺑ ﻲﺒﺳﺎﻨﻣ ﻱﺎﻫﺮﻛﺭﺎﻣ ،ﻲﺣﺍﻮﻧ ﻦﻳﺍ .(18) ﺩﻭﺭ ﺭﺎﻛ ﻪﺑ .ﺪﻨﺷﺎﺑﻲﻣ ﺎﻫﺎﻳﺰﺳﻻﺎﻣ ﻪﻠﻤﺟ ﺯﺍ ﺎﻫچﺭﺎﻗ ﺮﺘﺸﻴﺑ ﻚﻴﻣﻮﻧﻮﺴﻛﺎﺗ ﻩﺎﮕﻳﺎﺟ
ﻪﻴﺣﺎﻧ ﻦﻴﻤﻫ ﻑﺩﺍﺮﺗ ﺯﺍ ﻩﺩﺎﻔﺘﺳﺍ ﺎﺑ ﺭﺎﺑ ﻦﻴﻟﻭﺍ (9) ﺕﻮﻠﻳﻮﮔ ﻪﭽﻧﺎﻨﭼ
ﺪﻳﺪﺟﻪﻧﻮﮔ 4 ﺲﭙﺳ ﺩﻮﻤﻧﺩﺪﺠﻣ ﻱﺪﻨﺑﻪﻘﺒﻃﺍﺭ ﺎﻳﺰﺳﻻﺎﻣﻱﺎﻫﻪﻧﻮﮔ
ﻭ M. nana ،M. dermatis ،M. japonica ﻞﻣﺎﺷ ﺎﻳﺰﺳﻻﺎﻣ
ﻭ D1-D2-26SﻲﺣﺍﻮﻧﻲﻟﺍﻮﺗﻦﻴﻴﻌﺗﺯﺍﻩﺩﺎﻔﺘﺳﺍﺎﺑyM. yamatoensis
.(10-14) ﺩﺮﻛﻲﻓﺮﻌﻣ ITS1ﺎﻀﻌﺑ
ﺮﺑﻰﻨﺘﺒﻣPCR-RFLPﺖﺳﺍﺭﺮﺳ ﻭﻩﺩﺎﺳﻢﺘﺴﻴﺳﻚﻳﺶﻴﭘ ﺯﺍ
ﺎﻳﺰﺳﻻﺎﻣﻪﻧﻮﮔ 11 ﻲﻳﺎﺳﺎﻨﺷﻱﺍﺮﺑD1-D2 26S rDNAﻪﻴﺣﺎﻧﻰﻟﺍﻮﺗ
ﻱﺍﺮﺑﻢﺘﺴﻴﺳﻦﻳﺍ ﻪﻌﺳﻮﺗ ﻥﺎﻜﻣﺍ ﺵﺭﺍﺰﮔ ﻥﺎﻤﻫﺭﺩ ﻭ ﻢﻳﺩﻮﻤﻧ ﻲﻓﺮﻌﻣ
.(15) ﻢﻳﺩﺮﻛ ﺡﺮﻄﻣ ﺍﺭ ﺪﻳﺪﺟ ﻱﺎﻫﻪﻧﻮﮔ ﺖﻳﻮﻫ ﻦﻴﻴﻌﺗ ﻭ ﻲﻳﺎﺳﺎﻨﺷ
،ﺩﻮﻤﻧﻲﻓﺮﻌﻣﺎﻣﻪﺑﺍﺭﺪﻳﺪﺟﻪﻧﻮﮔﻲﺑﻮﺧﻪﺑﺭﻮﺑﺰﻣﻢﺘﺴﻴﺳﻪﻧﺎﺘﺨﺒﺷﻮﺧ ﻭﺖﺳﻮﭘﺭﺩﺩﻮﺟﻮﻣﺞﻳﺍﺭﻱﺎﻫﻪﻧﻮﮔﻲﻳﺎﺳﺎﻨﺷﻦﻴﺣﺭﺩﻪﻛﻱﺭﻮﻃﻪﺑ ﺲﭘﻭRFLPﺯﺍﻯﺍﻩﺯﺎﺗﻱﻮﮕﻟﺍﺎﺑﺩﺭﻮﺧﺮﺑﻦﻴﻟﻭﺍﺭﺩ،ﺕﺎﻧﺍﻮﻴﺣﻁﺎﺨﻣ
ﺎﻫﺎﻳﺰﺳﻻﺎﻣ ﺮﻳﺎﺳﺎﺑ ﻮﮕﻟﺍ ﻥﺁ ﻥﺩﻮﺑﺕﻭﺎﻔﺘﻣ ﺯﺍﻥﺎﻨﻴﻤﻃﺍ ﻭ ﻲﻫﺎﮔﺁ ﺯﺍ ﺎﺑ ﻪﻠﺌﺴﻣ ﻦﻳﺍ .ﺪﺷﺡﺮﻄﻣ ﺪﻳﺪﺟ ﻯﺍﻪﻟﻭﺰﻳﺍ ﻒﺸﻛ ﻥﺎﻜﻣﺍ ،(4 ﻞﻜﺷ)
ﻪﻴﺣﺎﻧﺰﻴﻧﻭ 26S rDNAﻪﻴﺣﺎﻧﺭﺩﺩﻮﺟﻮﻣ D1-D2ﻪﻴﺣﺎﻧﻲﻟﺍﻮﺗﻦﻴﻴﻌﺗ
ﻩﺩﺎﻔﺘﺳﺍﺎﺑ26Sﻰﻟﺍﻮﺗﺰﻴﻟﺎﻧﺁﺎﺑ .ﺪﻳﺩﺮﮔﺖﻳﻮﻘﺗﻭﺪﻴﻳﺎﺗﻲﺑﻮﺧﻪﺑITS1
ﻪﺑ RFLP ﺕﻻﻮﺼﺤﻣ ﻩﺯﺍﺪﻧﺍﻪﻛﺪﺷﻡﻮﻠﻌﻣDNASIS ﺭﺍﺰﻓﺍﻡﺮﻧ ﺯﺍ
ﻭﻪﻳﺎﭘﺭﺎﻴﻌﻣ .ﺪﺷﺎﺑﻰﻣﺯﺎﺑﺖﻔﺟ 61 ﻭ 70 ،143 ،297 ﻝﺩﺎﻌﻣﻖﻴﻗﺩﺭﻮﻃ
ﻪﻌﻄﻗﻑﺩﺍﺮﺗﺎﻳﺰﺳﻻﺎﻣﻯﺎﻫﻪﻧﻮﮔﻰﻣﻮﻧﻮﺴﻛﺎﺗﻭﻯﺪﻨﺑﻪﻘﺒﻃﻩﺪﺷﻪﺘﻓﺮﻳﺬﭘ ﻪﻧﻮﻤﻧﻞﻴﻟﺩﻦﻴﻤﻫﻪﺑ،(9-13) ﺪﺷﺎﺑﻰﻣ26S rDNAﻪﻴﺣﺎﻧﺭﺩD1/D2
ﺩﻮﺟﻭﻦﻳﺍﺎﺑ .ﺩﻮﺷﻰﻣﻰﻘﻠﺗﺪﻳﺪﺟﻯﺍﻪﻟﻭﺰﻳﺍ،ﻖﻴﻘﺤﺗﻦﻳﺍﻰﻃﻩﺪﺷﺍﺪﺟ
(Species) ﻪﻧﻮﮔﻥﺍﻮﻨﻋﻪﺑﻢﺴﻴﻧﺎﮔﺭﺍ ﻚﻳﻯﺭﺍﺬﮔﻡﺎﻧﻭﻰﻓﺮﻌﻣﻯﺍﺮﺑ
ﻚﻳژﻮﻟﻮﻳﺰﻴﻓ ﻭ ﻚﻳژﻮﻟﻮﻓﺭﻮﻣ ﺕﺎﺼﺨﺸﻣ ﻖﻴﻗﺩ ﺭﻮﻃ ﻪﺑ ﺖﺳﺍ ﻡﺯﻻ
ﻯﺭﺍﺪﻬﮕﻧ ﻰﻠﻠﻤﻟﺍﻦﻴﺑ ﺰﻛﺍﺮﻣ ﻪﺑ ﻩﺎﮕﻧﺁ ﻭ ﻩﺪﺷﻒﻴﺻﻮﺗ ﻭ ﻰﺳﺭﺮﺑ ﻥﺁ ﻡﺎﺠﻧﺍﻝﺎﺣﺮﻫﺭﺩﺕﺎﻌﻟﺎﻄﻣﻦﻳﺍ .ﺩﺩﺮﮔﺖﺒﺛﻭﻝﺎﺳﺭﺍﺎﻫﻢﺴﻴﻧﺎﮔﺭﺍﻭﺮﻜﻴﻣ .ﺖﺳﺍﺮﺘﺸﻴﺑﻯﺎﻫﻥﻮﻣﺯﺁﻭﻥﺎﻣﺯﻡﺰﻠﺘﺴﻣﻭﻩﺩﻮﺑ
PCR-RFLPﺯﺍﻞﺻﺎﺣﺞﻳﺎﺘﻧﺭﺩﻪﻛﻱﺪﻳﺪﺟﻱﻮﮕﻟﺍﻪﺑﻪﺟﻮﺗﺎﺑ
ﺲﻜﻠﭙﻤﻛ ﺭﺩ ﺩﻮﺟﻮﻣ ITS1 ﺰﻴﻧ ﻭ D1-D2 ﻲﺣﺍﻮﻧ ﻭ ﺪﺷ ﻩﺪﻫﺎﺸﻣ
ﻚﻴﻣﻮﻧﻮﺴﻛﺎﺗ ﻩﺎﮕﻳﺎﺟ ﻦﻴﻴﻌﺗ ﻱﺍﺮﺑ ﻲﺒﺳﺎﻨﻣ ﻱﺎﻫﺮﻛﺭﺎﻣ ،rDNA ﻰﻧژ
ﻱﺎﻳﺰﺳﻻﺎﻣﺭﺩﻰﺣﺍﻮﻧﻦﻳﺍ،ﻖﻴﻘﺤﺗﻦﻳﺍﻲﻃﺭﺩﻭﺪﻨﺷﺎﺑﻲﻣﺎﻫﺎﻳﺰﺳﻻﺎﻣ ﺩﺮﻓﻪﺑﺮﺼﺤﻨﻣﻯﺪﻴﺗﻮﺌﻠﻛﻮﻧﻰﻟﺍﻮﺗﻯﺍﺭﺍﺩﻯﺪﻨﻫﻪﭽﻛﻮﺧﺯﺍﻩﺪﺷﺍﺪﺟ ﻰﻓﺮﻌﻣ .ﺩﻮﺷﻲﻣﻲﻓﺮﻌﻣﺪﻳﺪﺟﻯﺍﻪﻟﻭﺰﻳﺍﻚﻳﻥﺍﻮﻨﻋﻪﺑﻪﻧﻮﻤﻧﻦﻳﺍ،ﻩﺩﻮﺑ
ﻒﻴﺻﻮﺗﻡﺰﻠﺘﺴﻣ،(Species) ﻪﻧﻮﮔﻥﺍﻮﻨﻋﻪﺑﻪﻟﻭﺰﻳﺍﻦﻳﺍﻯﺭﺍﺬﮔﻡﺎﻧﻭ
ﻪﺑﻥﺁﺖﺒﺛﻭﻝﺎﺳﺭﺍﺰﻴﻧﻭﻥﺁﻚﻳژﻮﻟﻮﻳﺰﻴﻓﻭﻚﻳژﻮﻟﻮﻓﺭﻮﻣﺕﺎﺼﺨﺸﻣ
.ﺖﺳﺎﻫﻢﺴﻴﻧﺎﮔﺭﺍﻭﺮﻜﻴﻣ ﻯﺭﺍﺪﻬﮕﻧﻰﻠﻠﻤﻟﺍﻦﻴﺑﺰﻛﺍﺮﻣ
ﺮﻜﺸﺗ
ﻭ
ﺮﻳﺪﻘﺗ
ﻲﻜﺷﺰﭘ ﻡﻮﻠﻋ ﻩﺎﮕﺸﻧﺍﺩ ﺕﺍﺭﺎﺒﺘﻋﺍ ﺯﺍ ﻩﺩﺎﻔﺘﺳﺍ ﺎﺑ ﻪﻌﻟﺎﻄﻣ ﻦﻳﺍ ﻱﺎﻫﻚﻤﻛ ﺯﺍ .ﺖﺳﺍ ﻩﺪﺷ ﻡﺎﺠﻧﺍ ﻲﻣﻼﺳﺍ ﺩﺍﺯﺁ ﻩﺎﮕﺸﻧﺍﺩ ﻭ ﻥﺍﺮﻬﺗ
ﻱﺭﺍﺰﮕﺳﺎﭙﺳﻪﻧﺎﻤﻴﻤﺻ ﻱﺮﻔﻌﺟﺎﺿﺭﻱﺎﻗﺁﻭﺪﻧﺯﻲﻟﻼﺟﺮﻓﻮﻠﻴﻧ ﻢﻧﺎﺧ
.ﺩﺩﺮﮔﻲﻣ
New Malassezia AJ249956M.sloofiae AF063214M.funfur
AB125263M.yamatoerisis AB100599M.japonica
AJ249950M.restricta
AF064024M.sympodialis AB075224M.nana Phylogram
AB070361M.dermatis AJ249951M.globosa AF063215M.pachydermatis
AJ249954M.obtusa
ﺎﻬﻧﺁﻦﻴﺑﺭﺩﺪﻳﺪﺟﻱﺎﻳﺰﺳﻻﺎﻣﻩﺎﮕﻳﺎﺟﻭﺎﻳﺰﺳﻻﺎﻣﻒﻠﺘﺨﻣﻱﺎﻫﻪﻧﻮﮔD1-D2ﻪﻴﺣﺎﻧﻡﺍﺮﮔﻮﻠﻳﺎﻓ :5 ﻞﻜﺷ
ﺎﻬﻧﺁﻦﻴﺑﺭﺩﺪﻳﺪﺟﻱﺎﻳﺰﺳﻻﺎﻣﻩﺎﮕﻳﺎﺟﻭﺎﻳﺰﺳﻻﺎﻣﻒﻠﺘﺨﻣﻱﺎﻫﻪﻧﻮﮔITS1ﻪﻴﺣﺎﻧﻡﺍﺮﮔﻮﻠﻳﺎﻓ :6 ﻞﻜﺷ
New Malassezia AB019348M.Slooffiae
AB019337M.pachydermatis
AB19336M.obtusa
AB075225M.nana
AB070356M.dermatis AB019345M.sympodialis AB125266M.yamatoensis
AB019329M.furfur
AB019340M.restricta
AB19342M.globosa AB19336M.obtusa
References
1. Ashbee HR. Update on the genus Malassezia, Med Mycol. 2007; 45(4): 287-303.
2. Chen TA, Hill PB. The biology of Malassezia organ-isms and their ability to induce immune responses and skin disease. Vet Dermatol. 2005; 16(1): 4-26.
3. Gupta AK, Batra R, Boekhout T. Skin diseases associ-ated with Malassezia species. J Am Acad of Dermatol. 2004; 51(5): 785-789.
4. Crespo Erchiga V, Florencio VD. Malassezia species in skin diseases. Curr Opin Infect Dis. 2002; 15(2): 133-142.
5. Faergemann J. Atopic dermatitis and fungi. Clin Micro-biol Rev. 2002; 15: 545-563.
6. Chryssanthou E, Broberger U, Petrini B. Chryssan-thou E, Broberger U, Petrini B. Malassezia pachyder-matis fungemia in a neonatal intensive care unit. Acta Paediatr. 2001; 90(3): 323-327.
7. Ashbee HR, Evans EG. Immunology of diseases as-sociated with Malassezia species. Clin Microbi Rev. 2002; 15(1): 1-27.
8. Simmons RB, Guého E. A new species of Malassezia. Mycol Res. 199b; 94: 1146-1149.
9. Guillot J, Guého E. The diversity of Malassezia yeasts confirmed by rRNA sequence and nuclear DNA compari-sons. Antonie Van Leeuwenhoek. 1995; 67(3): 297-314. 10. Hirai A, Kano R, Makimura K. Malassezia nana sp. nov., a novel lipid-dependent yeast species isolated from animals. Int J Sys Evol Microbiol. 2004; 54: 623-662. 11. Sugita T, Takashima M, Shinoda T. New yeast spe-cies, Malassezia dermatis, isolated from patients with at-opic dermatitis. J Clin Microbiol. 2003; 40(4): 1363-1367.
12. Sugita T, Takashima M, Kodama M. Description of a New Yeast Species, Malassezia japonica, and its detec-tion in patients with atopic dermatitis and healthy sub-jects. J Clin Microbiol. 2003; 41(10): 4695-4699. 13. Sugita T, Tajima M, Takashima M. A new yeast, Malassezia yamatoensis, Isolated from a Patient with Seborrheic Dermatitis, and Its Distribution in patients and Healthy Subjects. Microbiol Immunol. 2004; 48(8): 579-583.
14. Leeming JP, Notman FH, Holland KT. The distribu-tion and ecology of Malassezia furfur and cutaneous bacteria on human skin. J Appl Bacteriol. 1989; 67(1): 47-52.
15. Mirhendi H, Makimura K, Zomorodian K, Yamada T, Sugita T, Yamaguchi H. A simple PCR-RFLP method for identification and differentiation of 11 Malassezia spe-cies. J Microbiol Methods. 2005; 61(2): 281-284. 16. Yamada Y, Makimura K, Merhendi H, Ueda K, Nishi-yama Y, Yamaguchi H, et al. Comparison of different methods for extraction of mitochondrial DNA from hu-man pathogenic yeasts. Jpn J Infect Dis. 2002; 55(4): 122-125.
17. Makimura K, Tamura Y, Kudo M, Uchida K, Saito H, Yamaguchi H. Species identification and strain typing of Malassezia species stock strains and clinical isolates based on the DNA sequences of nuclear ribosomal in-ternal transcribed spacer 1 regions. J Med Microbiol. 2000; 49(1): 29-35.
