Use of a synthetic oligonucleotide to detect false positives caused by cross-contamination in nested PCR

IF 1.7 4区 生物学 Q4 BIOCHEMICAL RESEARCH METHODS Journal of microbiological methods Pub Date : 2024-09-11 DOI:10.1016/j.mimet.2024.107040
Alexandre S. Maekawa , Luciene S. Santos , Paulo E.N.F. Velho , Marina R. Drummond
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Abstract

Nested PCR is a useful tool for identifying low-abundance target sequences of pathogens and avoiding false negatives. However, it carries an increased risk of cross-contamination, especially with its positive control. Here, we propose using customized synthetic oligonucleotides to detect false positives due to cross-contamination.

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使用合成寡核苷酸检测嵌套 PCR 中交叉污染导致的假阳性结果
巢式 PCR 是鉴定病原体低丰度目标序列和避免假阴性的有效工具。然而,它也增加了交叉污染的风险,尤其是阳性对照。在此,我们建议使用定制的合成寡核苷酸来检测交叉污染导致的假阳性。
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来源期刊
Journal of microbiological methods
Journal of microbiological methods 生物-生化研究方法
CiteScore
4.30
自引率
4.50%
发文量
151
审稿时长
29 days
期刊介绍: The Journal of Microbiological Methods publishes scholarly and original articles, notes and review articles. These articles must include novel and/or state-of-the-art methods, or significant improvements to existing methods. Novel and innovative applications of current methods that are validated and useful will also be published. JMM strives for scholarship, innovation and excellence. This demands scientific rigour, the best available methods and technologies, correctly replicated experiments/tests, the inclusion of proper controls, calibrations, and the correct statistical analysis. The presentation of the data must support the interpretation of the method/approach. All aspects of microbiology are covered, except virology. These include agricultural microbiology, applied and environmental microbiology, bioassays, bioinformatics, biotechnology, biochemical microbiology, clinical microbiology, diagnostics, food monitoring and quality control microbiology, microbial genetics and genomics, geomicrobiology, microbiome methods regardless of habitat, high through-put sequencing methods and analysis, microbial pathogenesis and host responses, metabolomics, metagenomics, metaproteomics, microbial ecology and diversity, microbial physiology, microbial ultra-structure, microscopic and imaging methods, molecular microbiology, mycology, novel mathematical microbiology and modelling, parasitology, plant-microbe interactions, protein markers/profiles, proteomics, pyrosequencing, public health microbiology, radioisotopes applied to microbiology, robotics applied to microbiological methods,rumen microbiology, microbiological methods for space missions and extreme environments, sampling methods and samplers, soil and sediment microbiology, transcriptomics, veterinary microbiology, sero-diagnostics and typing/identification.
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