{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,2,21]],"date-time":"2025-02-21T14:08:53Z","timestamp":1740146933226,"version":"3.37.3"},"reference-count":4,"publisher":"Public Library of Science (PLoS)","issue":"12","license":[{"start":{"date-parts":[[2024,12,9]],"date-time":"2024-12-09T00:00:00Z","timestamp":1733702400000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/100006492","name":"Division of Intramural Research, National Institute of Allergy and Infectious Diseases","doi-asserted-by":"publisher","award":["R01AI177637"],"award-info":[{"award-number":["R01AI177637"]}],"id":[{"id":"10.13039\/100006492","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["www.plosone.org"],"crossmark-restriction":false},"short-container-title":["PLoS ONE"],"abstract":"<jats:p>Identifying pathogens, resistance-conferring mutations, and strain types through targeted amplicon sequencing is an important tool. However, due to the limitations of short read sequencing, many applications require the division of limited clinical samples. Here, we present stilPCR (single-tube Illumina long read PCR), which allows the generation of hemi-nested amplicons in a single tube, with Illumina indexes and adapters, effectively increasing the Illumina read length without increasing the input requirements of reagents or sample. We have successfully utilized stilPCR on clinical sputum from tuberculosis patients to detect drug resistance mutations.<\/jats:p>","DOI":"10.1371\/journal.pone.0314265","type":"journal-article","created":{"date-parts":[[2024,12,9]],"date-time":"2024-12-09T18:30:31Z","timestamp":1733769031000},"page":"e0314265","update-policy":"https:\/\/doi.org\/10.1371\/journal.pone.corrections_policy","source":"Crossref","is-referenced-by-count":0,"title":["stilPCR increases the effective sequencing length of Illumina targeted next-generation sequencing"],"prefix":"10.1371","volume":"19","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-7893-8416","authenticated-orcid":true,"given":"Jason D.","family":"Limberis","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Roland J.","family":"Nagel","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Soumitesh","family":"Chakravorty","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Dena","family":"Block","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Scott","family":"Dewell","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Alina","family":"Nalyvayko","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Zach","family":"Howard","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Grant","family":"Theron","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Rouxjeane","family":"Venter","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"John Z.","family":"Metcalfe","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"340","published-online":{"date-parts":[[2024,12,9]]},"reference":[{"key":"pone.0314265.ref001","article-title":"SemiQuant\/stilPCR: v1.0.1","author":"JD Limberis","year":"2024","journal-title":"Zenodo"},{"key":"pone.0314265.ref002","doi-asserted-by":"crossref","first-page":"422","DOI":"10.5588\/ijtldopen.24.0124","article-title":"Cartridge-based sequencing for bedaquiline resistance detection from sputum.","volume":"1","author":"JD Limberis","year":"2024","journal-title":"IJTLD OPEN"},{"key":"pone.0314265.ref003","doi-asserted-by":"crossref","first-page":"e604","DOI":"10.1016\/S2666-5247(21)00175-0","article-title":"Genetic variants and their association with phenotypic resistance to bedaquiline in Mycobacterium tuberculosis: a systematic review and individual isolate data analysis","volume":"2","author":"N Ismail","year":"2021","journal-title":"Lancet Microbe"},{"journal-title":"Catalogue of Mutations in Mycobacterium Tuberculosis Complex and Their Association with Drug Resistance Second Edition","year":"2023","key":"pone.0314265.ref004"}],"container-title":["PLOS ONE"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/dx.plos.org\/10.1371\/journal.pone.0314265","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2024,12,9]],"date-time":"2024-12-09T18:30:34Z","timestamp":1733769034000},"score":1,"resource":{"primary":{"URL":"https:\/\/dx.plos.org\/10.1371\/journal.pone.0314265"}},"subtitle":[],"editor":[{"given":"Nils","family":"Pilotte","sequence":"first","affiliation":[],"role":[{"role":"editor","vocabulary":"crossref"}]}],"short-title":[],"issued":{"date-parts":[[2024,12,9]]},"references-count":4,"journal-issue":{"issue":"12","published-online":{"date-parts":[[2024,12,9]]}},"URL":"https:\/\/doi.org\/10.1371\/journal.pone.0314265","relation":{},"ISSN":["1932-6203"],"issn-type":[{"type":"electronic","value":"1932-6203"}],"subject":[],"published":{"date-parts":[[2024,12,9]]}}}