Kalantari Narges, Bayani Masomeh, Ghaffari Salman
Cellular and Molecular Biology Research Center (CMBRC), Babol University of Medical Sciences, Babol, Iran. ; Department of Laboratory Sciences, Faculty of Para-Medicine, Babol University of Medical Sciences, Babol, Iran.
Infectious Diseases and Tropical Medicine Research Center, Babol University of Medical Sciences, Babol, Iran.
Int J Mol Cell Med. 2013 Summer;2(3):125-30.
Sarcocystis is a genus of cyst-forming parasites infecting both animals and human. This study aimed to isolate coccidian tissue cysts from muscle of infected animals by a simple method in addition to molecular identification of Sarcocystis cruzi from the samples. The samples were obtained from commercial source in Babol, Northern Iran. Five grams of calf muscle was cut into 2-3 pieces in 30 ml of phosphate-buffered saline containing 0.1% Tween 80 and homogenized with IKA T25, DIGITAL ULTRA-TURRAX. The homogenate was filtrated twice and used for microscopy examination and molecular analysis. Polymerase chain reaction (PCR) and partial sequence analysis of the 18S ribosomal gene were used to identify the Sarcocystis species. Giemsa stain of the filtrated calf muscle samples showed that the sample had ellipse to around tissue cysts contained crescent-shaped bodies. The PCR of the 18SrDNA yielded an 1100 bp DNA band on agarose gel and sequence analysis of the DNA confirmed the isolate as S. cruzi. The sequence was deposited in GenBank by Accession No.KC508514. This is the first molecular identification of an isolate of S. cruzi in Iran.
肉孢子虫是一种形成包囊的寄生虫属，可感染动物和人类。本研究旨在通过一种简单方法从受感染动物的肌肉中分离出球虫组织包囊，并对样本中的克鲁斯肉孢子虫进行分子鉴定。样本取自伊朗北部巴博勒的商业来源。将5克小牛肌肉切成2 - 3块，放入30毫升含0.1%吐温80的磷酸盐缓冲盐水中，用IKA T25数字超高速均质器进行均质处理。匀浆过滤两次后用于显微镜检查和分子分析。采用聚合酶链反应（PCR）和18S核糖体基因部分序列分析来鉴定肉孢子虫种类。对过滤后的小牛肌肉样本进行吉姆萨染色显示，样本中有椭圆形至圆形的组织包囊，内含月牙形物体。18SrDNA的PCR在琼脂糖凝胶上产生了一条1100 bp的DNA条带，DNA序列分析证实该分离株为克鲁斯肉孢子虫。该序列已以登录号KC508514存入GenBank。这是伊朗首次对克鲁斯肉孢子虫分离株进行分子鉴定。
