Department of Bacteriology, Mycology and Parasitology, Statens Serum Institut, Artillerivej 5, DK-2300 Copenhagen S, Denmark.
Mol Cell Probes. 2010 Dec;24(6):403-6. doi: 10.1016/j.mcp.2010.07.008. Epub 2010 Aug 4.
A method using a single-round PCR coupled to pyrosequencing was developed for the detection and differentiation of members of the Entamoeba complex. The technique was evaluated using DNA isolated directly from faecal specimens and compared with a duplex real-time PCR targeting Entamoeba histolytica and Entamoeba dispar, and a conventional single-round PCR for the detection of Entamoeba moshkovskii. Tetranucleate cysts from 102 faecal specimens from Swedish, Danish and Dutch patients test-positive for the Entamoeba complex by coproscopic examination were identified to species using each of the three methods. Although none of the patients were confirmed to be positive for E. moshkovskii, E. histolytica and E. dispar were identified in 17 and 86 of the samples, respectively, one of the samples containing both species. There was concordance in results between pyrosequencing and the two other methods used. This study showed that PCR and pyrosequencing could be used for the rapid and high throughput identification of Entamoeba species.
一种使用单轮 PCR 结合焦磷酸测序的方法被开发出来，用于检测和区分肠贾第虫复合体的成员。该技术使用直接从粪便标本中分离的 DNA 进行评估，并与靶向溶组织内阿米巴和迪斯帕内阿米巴的双实时 PCR 以及用于检测莫氏内阿米巴的常规单轮 PCR 进行比较。使用这三种方法中的每一种，对来自瑞典、丹麦和荷兰患者的 102 份粪便标本中通过粪便镜检呈肠贾第虫复合体阳性的四核包囊进行了鉴定。虽然没有患者被确认为莫氏内阿米巴阳性，但在 17 份和 86 份样本中分别鉴定出了溶组织内阿米巴和迪斯帕内阿米巴，其中一份样本同时含有两种物种。PCR 和焦磷酸测序的结果与使用的另外两种方法一致。本研究表明，PCR 和焦磷酸测序可用于快速高通量鉴定内阿米巴物种。
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