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

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. Created with BioRender.com. [Display omitted] •Alternative nested PCR positive control to minimize false positives.•New method to detect cross-contamination in nucleic acid amplification procedures.•Customized oligonucleotide for nested PCR to detect Bartonella Henselae.