Zebardast Nozhat, Haghighi Ali, Yeganeh Farshid, Seyyed Tabaei Seyyed Javad, Gharavi Mohammad Javad, Fallahi Shirzad, Lasjerdi Zohreh, Salehi Nima, Taghipour Niloofar, Kohansal Cobra, Naderi Farideh
Dept. of Parasitology and Mycology, Shahid Beheshti University of Medical Sciences, Tehran, Iran.
Dept. of Immunology, Shahid Beheshti University of Medical Sciences, Tehran, Iran.
Iran J Parasitol. 2014 Oct-Dec;9(4):466-73.
Entamoeba moshkovskii and E. dispar are impossible to differentiate microscopically from the pathogenic species E. histolytica. Multiplex polymerase chain reaction (Multiplex PCR) is a widespread molecular biology technique for amplification of multiple targets in a single PCR experiment.
For detection and differentiation of the three-microscopy indistinguishable Entamoeba species in human, multiplex PCR assay using different DNA extraction methods was studied. A conserved forward primer was derived from the middle of the small-subunit rRNA gene, and reverse primers were designed from signature sequences specific to each of these three Entamoeba species.
A 166-bp PCR product with E. histolytica DNA, a 580-bp product with E. moshkovskii DNA and a 752-bp product with E. dispar DNA were generated in a single-round and multiplex PCR reaction.
We recommend this PCR assay as an accurate, rapid, and effective diagnostic method for the detection and discrimination of these three Entamoeba species in both routine diagnosis of amoebiasis and epidemiological surveys.
莫氏内阿米巴和迪斯帕内阿米巴在显微镜下无法与致病的溶组织内阿米巴区分开来。多重聚合酶链反应（多重PCR）是一种广泛应用的分子生物学技术，可在单个PCR实验中扩增多个靶标。
为检测和区分人类中三种在显微镜下无法区分的内阿米巴物种，研究了使用不同DNA提取方法的多重PCR检测方法。一个保守的正向引物来自小亚基rRNA基因的中部，反向引物则根据这三种内阿米巴物种各自的特征序列设计。
在单轮多重PCR反应中，产生了一条166bp的溶组织内阿米巴DNA的PCR产物、一条580bp的莫氏内阿米巴DNA的产物和一条752bp的迪斯帕内阿米巴DNA的产物。
我们推荐这种PCR检测方法，作为在阿米巴病的常规诊断和流行病学调查中检测和鉴别这三种内阿米巴物种的准确、快速且有效的诊断方法。
