Nowzari N, Djadid N Dinparast, Rahbari S, Yakchali B, Kazemi B, Jula G Moazeni
Veterinary Organization of Iran, P.O. Box 14155-6349, Tehran, Iran.
Vet Parasitol. 2005 Mar 10;128(1-2):59-64. doi: 10.1016/j.vetpar.2004.11.015. Epub 2004 Dec 25.
Eimeria species from poultry breeder farms without previous exposure to anticoccidial vaccines in five distinct geographical regions of Iran were examined for genetic relatedness by the random amplified polymorphic DNA (RAPD) assay. Eight different oligonucleotide decamers with arbitrary DNA sequences were tested as primers to amplify DNA from five isolates of each E. acervulina, E. tenella, and E. maxima. Depending on the species/isolate-primer combination, between 1 and 14 DNA fragments ranging in size from 240 to 3000 bp were amplified. The two isolates originated from Northeast and North parts of Iran showed minor differences and two isolates originated from Northeast and Southwest of Iran showed major differences in their amplified DNA patterns. The intra-specific similarity coefficient within five isolates of each species of, E. acervulina, E. tenella, E. maxima was 74, 82 and 72%, respectively. The distance indices observed between species were greater than those found between isolates (80-90%) with all examined primers. The inferred phylogenetic tree on the fingerprinting of all species revealed that the RAPD-PCR can easily differentiate within and between species and could be a useful and valuable tool in future epidemiological studies, designing and developing of vaccines against avian coccidosis, here in Iran and neighboring countries.
采用随机扩增多态性DNA(RAPD)分析法,对来自伊朗五个不同地理区域、此前未接触过抗球虫疫苗的家禽种鸡场的艾美耳球虫进行遗传相关性检测。测试了八种具有任意DNA序列的不同寡核苷酸十聚体作为引物,以扩增来自堆型艾美耳球虫、柔嫩艾美耳球虫和巨型艾美耳球虫各五个分离株的DNA。根据物种/分离株 - 引物组合的不同,扩增出1至14个大小在240至3000 bp之间的DNA片段。源自伊朗东北部和北部的两个分离株在扩增的DNA图谱上显示出微小差异,而源自伊朗东北部和西南部的两个分离株则显示出较大差异。堆型艾美耳球虫、柔嫩艾美耳球虫、巨型艾美耳球虫各五个分离株的种内相似系数分别为74%、82%和72%。在所检测的所有引物中,观察到的种间距离指数大于分离株间的距离指数(80 - 90%)。根据所有物种指纹图谱推断的系统发育树表明,RAPD-PCR能够轻松区分种内和种间差异,并且在伊朗及周边国家未来的流行病学研究、抗禽球虫病疫苗的设计与开发中可能是一种有用且有价值的工具。