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 posit...
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| Veröffentlicht in: | Journal of microbiological methods 2024-11, Vol.226, p.107040, Article 107040 |
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| container_title | Journal of microbiological methods |
| container_volume | 226 |
| creator | Maekawa, Alexandre S. Santos, Luciene S. Velho, Paulo E.N.F. Drummond, Marina R. |
| description | 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. |
| doi_str_mv | 10.1016/j.mimet.2024.107040 |
| format | Article |
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Created with BioRender.com. [Display omitted]
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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.</description><subject>Bartonella henselae</subject><subject>Contamination detection</subject><subject>False-positive</subject><subject>Molecular diagnostics</subject><subject>Nested PCR</subject><subject>Oligonucleotide</subject><issn>0167-7012</issn><issn>1872-8359</issn><issn>1872-8359</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2024</creationdate><recordtype>article</recordtype><recordid>eNp9kMtKAzEUhoMoWqtPIEiWbqbmMrcsXEjxBoIidh0yyRlNmUnqJFPo2xvb6tJVDuH7z598CF1QMqOEltfLWW97iDNGWJ5uKpKTAzShdcWymhfiEE0SVWUVoewEnYawJIQWPK-P0QkXrCJVQSfocxEA-xYrHDYufkK0GvvOfng36g58tAZw9NhABB1xq7qEr3yw0a4hYK3GAAY3G6wHH0KmvYuqt05F6x22DjsIMQGv87czdLRNn-_PKVrc373PH7Pnl4en-e1zphkXMWuawtQla4RplFBQcsY5pFm10OaUkFy0DZiCi1q1FEQllFK8ZaIkAIrxmk_R1W7vavBfY6qXvQ0auk458GOQnJK84GUhaEL5Dt0-foBWrgbbq2EjKZE_iuVSbhXLH8VypzilLvcFY9OD-cv8Ok3AzQ6A9M21hUEGbcFpMHZIEqXx9t-Cb-CDkAc</recordid><startdate>20241101</startdate><enddate>20241101</enddate><creator>Maekawa, Alexandre S.</creator><creator>Santos, Luciene S.</creator><creator>Velho, Paulo E.N.F.</creator><creator>Drummond, Marina R.</creator><general>Elsevier B.V</general><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>7X8</scope></search><sort><creationdate>20241101</creationdate><title>Use of a synthetic oligonucleotide to detect false positives caused by cross-contamination in nested PCR</title><author>Maekawa, Alexandre S. ; Santos, Luciene S. ; Velho, Paulo E.N.F. ; Drummond, Marina R.</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c239t-bb5d862b9dba9ae63233edbaafef410049fbed5398af1e979aaa3f2960eea2383</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2024</creationdate><topic>Bartonella henselae</topic><topic>Contamination detection</topic><topic>False-positive</topic><topic>Molecular diagnostics</topic><topic>Nested PCR</topic><topic>Oligonucleotide</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Maekawa, Alexandre S.</creatorcontrib><creatorcontrib>Santos, Luciene S.</creatorcontrib><creatorcontrib>Velho, Paulo E.N.F.</creatorcontrib><creatorcontrib>Drummond, Marina R.</creatorcontrib><collection>PubMed</collection><collection>CrossRef</collection><collection>MEDLINE - Academic</collection><jtitle>Journal of microbiological methods</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Maekawa, Alexandre S.</au><au>Santos, Luciene S.</au><au>Velho, Paulo E.N.F.</au><au>Drummond, Marina R.</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Use of a synthetic oligonucleotide to detect false positives caused by cross-contamination in nested PCR</atitle><jtitle>Journal of microbiological methods</jtitle><addtitle>J Microbiol Methods</addtitle><date>2024-11-01</date><risdate>2024</risdate><volume>226</volume><spage>107040</spage><pages>107040-</pages><artnum>107040</artnum><issn>0167-7012</issn><issn>1872-8359</issn><eissn>1872-8359</eissn><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.
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.</abstract><cop>Netherlands</cop><pub>Elsevier B.V</pub><pmid>39270751</pmid><doi>10.1016/j.mimet.2024.107040</doi></addata></record> |
| fulltext | fulltext |
| identifier | ISSN: 0167-7012 |
| ispartof | Journal of microbiological methods, 2024-11, Vol.226, p.107040, Article 107040 |
| issn | 0167-7012 1872-8359 1872-8359 |
| language | eng |
| recordid | cdi_proquest_miscellaneous_3104536591 |
| source | Elsevier ScienceDirect Journals |
| subjects | Bartonella henselae Contamination detection False-positive Molecular diagnostics Nested PCR Oligonucleotide |
| title | Use of a synthetic oligonucleotide to detect false positives caused by cross-contamination in nested PCR |
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