用RAPD标记检测棘阿米巴基因组DNA多态性

本文用ＲＡＰＤ技术就我国首次分离到的棘阿米巴角膜炎病原体Ｗ株（ＥＣＮＵ－９２Ｗ１），同其它４个虫株（Ａｃａｎｔｈａｍｏｅｂａｐｏｌｙｐｈａｇａ，Ａ．ｌｕｇｄｕｎｅｎｓｉｓ，Ａ．ｑｕｉｎａ，Ａ．ｐａｌｅｓｔｉｎｅｎｓｉｓ）进行了分子分类的比较研究，并对各虫株的系统发生进行了初步探讨。虫株间的共享度各有差异，其中Ａ．ｌｕｇｄｕｎｅｎｓｉｓ和Ａ．ｑｕｉｎａ最大，为０．６４５；Ｗ和Ａ．ｌｕｇｄｕｎｅｎｓｉｓ，Ａ．ｑｕｉｎａ分别为０．６３６，０．６００；Ａ．ｐｏｌｙｐｈａｇａ和Ａ．ｌｕｇｄｕｎｅｎｓｉｓ，Ａ．ｑｕｉｎａ及Ｗ分别为０．５７６，０．４７６，０．４７８；Ａ．ｐａｌｅｓｔｉｎｅｎｓｉｓ和Ａ．ｐｏｌｙｐｈａｇａ，Ａ．ｌｕｇｄｕｎｅｎｓｉｓ，Ａ．ｑｕｉｎａ及Ｗ分别为０．２２５，０．２００，０．２７０，０．３０８。结合以往对形态学，同工酶和ｍｔＤＮＡ限制性内切酶分析的结果，进一步为Ａ．ｌｕｇｄｕｎｅｎｓｉｓ－Ａ．ｑｕｉｎａｃｏｍｐｌｅｘ提供了分子证据，并将Ｗ株初步鉴定为此复合体

DETECTION OF ACANTHAMOEBA GENOMIC DNA POLYMORPHISMS USING RAPD MOLECULAR MARKERS *

The pathogenic strain W(ECNU 92W1) was first isolated from an acanthamoeba keratitis patient wearing soft contact lens in China.Genomic DNA polymorphisms within the five Acantahamoeba strains/isolate, A.polyphaga (ATCC30462), A.lugdunensis (ATCC 50240), A.quina (ATCC 50241), A.palestinensis (ATCC 30870) and W were detected using random amplified polymorphic DNA (RAPD) markers with nine different 10 mer primers of arbitrary nucleotide sequence in the present work.The experimental results showed that each strain had its characteristic patterns and there were some differences of the shared random amplified DNA fragments between these strains.The molecular data were provided for considering A.lugdunensis and A.quina as a complex,and the Chinese isolate W was preliminary identified as a member of the A.lugdunensis A.quina complex.Some systematic problems of genus Acanthamoeba were discussed.