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物种特异性重复DNA序列的自动识别及其在检测微生物中的应用。

Automatic identification of species-specific repetitive DNA sequences and their utilization for detecting microbial organisms.

作者信息

Koressaar Triinu, Jõers Kai, Remm Maido

机构信息

Department of Bioinformatics, Institute of Molecular and Cell Biology, University of Tartu, Tartu, Estonia.

出版信息

Bioinformatics. 2009 Jun 1;25(11):1349-55. doi: 10.1093/bioinformatics/btp241. Epub 2009 Apr 8.

Abstract

MOTIVATION

The concentration of pathogen DNA in biological samples is often very low. Therefore, the sensitivity of diagnostic tests is always a critical factor.

RESULTS

We have developed a novel computational method that identifies species-specific repeats from microbial organisms and automatically designs species-specific PCR primers for these repeats. We tested the methodology on 30 randomly chosen microbial species and we demonstrate that species-specific repeats longer than 300 bp exist in all these genomes. We also used our methodology to design species-specific PCR primers for 86 repeats from five medically relevant microbial species. These PCR primers were tested experimentally. We demonstrate that using species-specific repeats as a PCR template region can increase the sensitivity of PCR in diagnostic tests.

AVAILABILITY AND IMPLEMENTATION

A web version of the method called MultiMPrimer3 was implemented and is freely available at (http://bioinfo.ut.ee/multimprimer3/).

摘要

动机

生物样本中病原体DNA的浓度通常非常低。因此,诊断测试的灵敏度始终是一个关键因素。

结果

我们开发了一种新颖的计算方法,该方法可从微生物中识别物种特异性重复序列,并自动为这些重复序列设计物种特异性PCR引物。我们在30个随机选择的微生物物种上测试了该方法,结果表明所有这些基因组中都存在长度超过300 bp的物种特异性重复序列。我们还使用我们的方法为来自五个医学相关微生物物种的86个重复序列设计了物种特异性PCR引物。对这些PCR引物进行了实验测试。我们证明,使用物种特异性重复序列作为PCR模板区域可以提高诊断测试中PCR的灵敏度。

可用性和实现方式

实现了该方法的网络版本,称为MultiMPrimer3,可在(http://bioinfo.ut.ee/multimprimer3/)上免费获取。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/4bc1/2682524/111afb6e5d33/btp241f1.jpg

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