Sarkari Shahriari Bahador, Biranvand Elahe, Sadjjadi Seyed Mahmoud, Rahimi Hamidreza
Basic Sciences in Infectious Diseases Research Center, Shiraz University of Medical Sciences, Shiraz, Iran.
Dept. of Parasitology and Mycology, School of Medicine, Shiraz University of Medical Sciences, Shiraz, Iran.
Iran J Parasitol. 2013 Oct;8(4):545-51.
Antigen B (AgB) is frequently used for immuno-diagnosis of human cystic echinococcosis (CE). Echinococcus granulosus AgBs show a high degree of genetic variability in different hosts or in different CE endemic areas. The present study aimed to evaluate the genetic polymorphisms of encoding antigen B2 gene (AgB2) among different Iranian isolates of E. granulosus.
A total of 50 CE isolates were collected from human, sheep, cattle, goat and camels, 10 isolates from each intermediate host of E. granulosus. Total genomic DNA from either protoscolices or germinal layer was extracted from each cyst and PCR-RFLP followed by DNA sequencing was used to evaluate sequence variation and polymorphism of AgB2 in the isolates.
After the PCR amplification, using AgB2 primers, an almost 400 bp band was amplified in all of the isolates. The PCR products were digested with Alu1 restriction endonuclease. After restriction enzyme digestion with Alu1' sheep and human isolates gave a similar pattern of RFLP with the gene size of approximately 140 and 240bp and camel and goat isolates gave a similar pattern, but different from sheep and human, with the gene size of approximately 150 and 250bp. Sequence analysis showed the most genetic similarity of AgB2 between human and sheep isolates.
Findings of this study revealed the differences in the sequences of AgB2 within and between the Iranian isolates of E. granulosus. These differences may affect the performance of any diagnostic test which uses AgB.
抗原B(AgB)常用于人类囊性棘球蚴病(CE)的免疫诊断。细粒棘球绦虫AgB在不同宿主或不同CE流行地区表现出高度的遗传变异性。本研究旨在评估伊朗不同细粒棘球绦虫分离株中编码抗原B2基因(AgB2)的遗传多态性。
共收集了50株CE分离株,分别来自人、绵羊、牛、山羊和骆驼,每种细粒棘球绦虫中间宿主各10株。从每个囊肿的原头节或生发层中提取总基因组DNA,采用PCR-RFLP并结合DNA测序来评估分离株中AgB2的序列变异和多态性。
使用AgB2引物进行PCR扩增后,所有分离株中均扩增出一条约400 bp的条带。PCR产物用Alu1限制性内切酶消化。用Alu1进行酶切后'绵羊和人分离株呈现相似的RFLP模式,基因大小约为140和240bp,骆驼和山羊分离株呈现相似模式,但与绵羊和人不同,基因大小约为150和250bp。序列分析显示人分离株和绵羊分离株之间AgB2的遗传相似性最高。
本研究结果揭示了伊朗细粒棘球绦虫分离株内部和之间AgB2序列的差异。这些差异可能会影响任何使用AgB的诊断测试的性能。