随机扩增多态性DNA图谱作为鉴定迪氏棘阿米巴的一种工具。

Random amplified polymorphic DNA profiles as a tool for the identification of Acanthamoeba divionensis.

作者信息

Ortega-Rivas Antonio, Lorenzo-Morales Jacob, Alonso Violeta, Abreu Néstor J, Foronda Pilar, del Castillo Antonio, Valladares Basilio

机构信息

Department of Parasitology, University of La Laguna, Avenida Astrofisico F, Sánchez S/N, ZC: 38.203, La Laguna, Canary Islands, Spain.

出版信息

Curr Microbiol. 2003 Aug;47(2):84-6. doi: 10.1007/s00284-002-3939-7.

DOI:10.1007/s00284-002-3939-7

PMID:14506852

Abstract

In the present study, we demonstrated the Random Amplified Polymorphism DNA (RAPD) diagnostic validity. In our study, we have analyzed RAPD profiles searching for characteristic and useful bands for Acanthamoeba diagnosis at the species level. We found a distinctive 370-bp band in A. divionensis RAPD patterns, using the OPC14 primer (TGCGTGCTTG) (Operon Technologies, Inc., Alameda, CA). The band specificity was confirmed by hybridization, using it as a probe, against all OPC14 amplifications from 10 different Acanthamoeba species. Once we sequenced this band, we used it to design a specific primer pair which showed positive amplification only in A. divionensis isolates.

摘要

在本研究中，我们证明了随机扩增多态性DNA（RAPD）的诊断有效性。在我们的研究中，我们分析了RAPD图谱，以寻找在物种水平上用于棘阿米巴诊断的特征性和有用条带。我们使用OPC14引物（TGCGTGCTTG）（Operon Technologies公司，阿拉米达，加利福尼亚州）在迪氏棘阿米巴的RAPD模式中发现了一条独特的370 bp条带。通过将其用作探针与来自10种不同棘阿米巴物种的所有OPC14扩增产物进行杂交，证实了该条带的特异性。一旦我们对这条带进行了测序，我们就用它设计了一对特异性引物，该引物仅在迪氏棘阿米巴分离株中显示出阳性扩增。