{"id":48,"date":"2013-07-17T17:19:51","date_gmt":"2013-07-17T21:19:51","guid":{"rendered":"https:\/\/sites.bu.edu\/biomol\/?page_id=48"},"modified":"2013-07-18T16:58:42","modified_gmt":"2013-07-18T20:58:42","slug":"pna-rca-fish","status":"publish","type":"page","link":"https:\/\/sites.bu.edu\/biomol\/projects\/pna-rca-fish\/","title":{"rendered":"PNA-RCA-FISH"},"content":{"rendered":"<p>We developed a robust DNA-targeted assay based on PNA technology that makes it possible to detect signature sequences on bacterial genomic DNA under non-denaturing, isothermal conditions within fixed cells.\u00a0This method provides us with single copy sensitivity and single base specificity due to a combination of three techniques: First, a peptide nucleic acid (PNA) probe locally opens a chosen target site, which allows a padlock DNA probe to access the site and become ligated. Second, rolling circle amplification (RCA) generates thousands of single-stranded copies of the target sequence. Finally, fluorescent\u00a0<i>in situ<\/i> hybridization (FISH) is used to visualize the amplified DNA.<\/p>\n<p>We have successfully used this approach to fluorescently detect single-copy DNA sequences in bacterial genomes and discriminate between similar sites in various bacteria based on single-nucleotide differences. In a collaborative effort with Boston Medical Center, we employed this technique to develope a novel diagnostic that can detect <i>Staphylococcus aureus <\/i>and simultaneously distinguish between methicillin-sensitive (MSSA) and methicillin-resistant (MRSA) strains.<\/p>\n","protected":false},"excerpt":{"rendered":"<p>We developed a robust DNA-targeted assay based on PNA technology that makes it possible to detect signature sequences on bacterial genomic DNA under non-denaturing, isothermal conditions within fixed cells.\u00a0This method provides us with single copy sensitivity and single base specificity due to a combination of three techniques: First, a peptide nucleic acid (PNA) probe locally [&hellip;]<\/p>\n","protected":false},"author":7350,"featured_media":0,"parent":104,"menu_order":3,"comment_status":"closed","ping_status":"closed","template":"","meta":[],"_links":{"self":[{"href":"https:\/\/sites.bu.edu\/biomol\/wp-json\/wp\/v2\/pages\/48"}],"collection":[{"href":"https:\/\/sites.bu.edu\/biomol\/wp-json\/wp\/v2\/pages"}],"about":[{"href":"https:\/\/sites.bu.edu\/biomol\/wp-json\/wp\/v2\/types\/page"}],"author":[{"embeddable":true,"href":"https:\/\/sites.bu.edu\/biomol\/wp-json\/wp\/v2\/users\/7350"}],"replies":[{"embeddable":true,"href":"https:\/\/sites.bu.edu\/biomol\/wp-json\/wp\/v2\/comments?post=48"}],"version-history":[{"count":11,"href":"https:\/\/sites.bu.edu\/biomol\/wp-json\/wp\/v2\/pages\/48\/revisions"}],"predecessor-version":[{"id":52,"href":"https:\/\/sites.bu.edu\/biomol\/wp-json\/wp\/v2\/pages\/48\/revisions\/52"}],"up":[{"embeddable":true,"href":"https:\/\/sites.bu.edu\/biomol\/wp-json\/wp\/v2\/pages\/104"}],"wp:attachment":[{"href":"https:\/\/sites.bu.edu\/biomol\/wp-json\/wp\/v2\/media?parent=48"}],"curies":[{"name":"wp","href":"https:\/\/api.w.org\/{rel}","templated":true}]}}