Ayyanathan K, Datta S
Astra Research Centre India, Malleswaram, Bangalore, India.
Mol Cell Probes. 1996 Aug;10(4):273-8. doi: 10.1006/mcpr.1996.0036.
DNA-based detection systems are being rapidly adapted for diagnostic technologies. The currently available non-radioactive DNA techniques for malaria detection are primarily based on probes which are species-specific. The major requirement of a cross-species (generic) diagnostics is the identification of an universal probe which is long enough to be used in either PCR amplification or solid state capture of the hybrids, the two principal techniques currently used in the field of non-radioactive DNA detection. This paper describes a DNA sequence (pARC 178), originally obtained from P. falciparum genomic library, which is also present in other malarial species. Southern-blot analysis indicates that this sequence is derived from a repetitive DNA element. This DNA fragment encodes for a small RNA species of approximately 120 bases. PCR primers based on this sequence amplifies the expected size (157 bp) product from the genomic DNA isolated from P. falciparum, P. vivax, P. berghei, P. yoelii, P. vinckei and P. chabaudi thus confirming its generic nature. The utility of this probe is further demonstrated by its capability in successfully detecting P. falciparum and P. vivax infections in clinical samples when used in a PCR assay.
基于DNA的检测系统正在迅速应用于诊断技术。目前可用于疟疾检测的非放射性DNA技术主要基于物种特异性探针。跨物种(通用)诊断的主要要求是鉴定一种通用探针,其长度要足够长,以便用于PCR扩增或杂交体的固态捕获,这是目前非放射性DNA检测领域使用的两种主要技术。本文描述了一个DNA序列(pARC 178),最初从恶性疟原虫基因组文库中获得,其他疟原虫物种中也存在该序列。Southern印迹分析表明该序列源自一个重复DNA元件。这个DNA片段编码一种约120个碱基的小RNA物种。基于该序列的PCR引物能从恶性疟原虫、间日疟原虫、伯氏疟原虫、约氏疟原虫、文氏疟原虫和查巴迪疟原虫分离的基因组DNA中扩增出预期大小(157 bp)的产物,从而证实了其通用性。当用于PCR检测时,该探针在成功检测临床样本中的恶性疟原虫和间日疟原虫感染方面的能力进一步证明了其效用。