{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,8,29]],"date-time":"2026-08-29T09:25:55Z","timestamp":1787995555589,"version":"build-2784847793"},"reference-count":39,"publisher":"American Chemical Society (ACS)","issue":"31","license":[{"start":{"date-parts":[[2026,7,29]],"date-time":"2026-07-29T00:00:00Z","timestamp":1785283200000},"content-version":"stm-asf","delay-in-days":0,"URL":"https:\/\/doi.org\/10.15223\/policy-029"},{"start":{"date-parts":[[2026,7,29]],"date-time":"2026-07-29T00:00:00Z","timestamp":1785283200000},"content-version":"stm-asf","delay-in-days":0,"URL":"https:\/\/doi.org\/10.15223\/policy-037"},{"start":{"date-parts":[[2026,7,29]],"date-time":"2026-07-29T00:00:00Z","timestamp":1785283200000},"content-version":"stm-asf","delay-in-days":0,"URL":"https:\/\/doi.org\/10.15223\/policy-045"}],"funder":[{"DOI":"10.13039\/501100001809","name":"National Natural Science Foundation of China","doi-asserted-by":"publisher","award":["22204109"],"award-info":[{"award-number":["22204109"]}],"id":[{"id":"10.13039\/501100001809","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100001809","name":"National Natural Science Foundation of China","doi-asserted-by":"publisher","award":["22325403"],"award-info":[{"award-number":["22325403"]}],"id":[{"id":"10.13039\/501100001809","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100004829","name":"Science and Technology Department of Sichuan Province","doi-asserted-by":"publisher","award":["2025NSFTD0001"],"award-info":[{"award-number":["2025NSFTD0001"]}],"id":[{"id":"10.13039\/501100004829","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"published-print":{"date-parts":[[2026,8,11]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                  <jats:p>Quantitative PCR (qPCR) is the gold standard for molecular diagnostics. Colorimetric readout can greatly simplify PCR-based diagnostics and make them appealing for decentralized testing. Current colorimetric PCR schemes mostly target the dsDNA amplicons. It is expected that pyrophosphate (Py), the inherent product of PCR (in fact, all nucleic acid amplification schemes), stands as a simpler colorimetric target, due to its versatile complexing with various metal ions\/indicators. However, Py-based colorimetric PCR has not been reported yet, partially because it will receive serious competitive complexation from Mg2+ in PCR. Herein, we introduced the Sc3+-hydroxynaphthol blue (Sc-HNB) complex to facilitate Py-based colorimetric PCRs. Because of the higher stability of Sc-HNB over Mg-HNB and Sc-Py over Mg-Py, Py generated during PCR cycles can directly displace HNB from the Sc-HNB complex in a Mg2+-independent manner, yielding a distinct color change. The proposed strategy features simplicity and low cost and can be coupled with PCR in both real-time and end-point manner, with sensitivity for target gene detection comparable to conventional qPCR. Because of the robustness of the Py-displacement reactions, the proposed colorimetric PCR could work with even water bath heating and was compatible with the existing PCR kits. To further validate its feasibility and accuracy, human papillomavirus (HPV) detection in vaginal swab samples was carried out, with results fully consistent with those obtained by qPCR. All of these results indicate that the Py-displacement-based colorimetric PCR can be a promising tool for decentralized nucleic acid testing.<\/jats:p>","DOI":"10.1021\/acs.analchem.6c01682","type":"journal-article","created":{"date-parts":[[2026,8,4]],"date-time":"2026-08-04T13:15:35Z","timestamp":1785849335000},"page":"22792-22799","source":"Crossref","is-referenced-by-count":1,"title":["Colorimetric\nPCR Based on Pyrophosphate Displacement"],"prefix":"10.1021","volume":"98","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-0332-0918","authenticated-orcid":true,"given":"Xianming","family":"Li","sequence":"first","affiliation":[{"name":"Sichuan University , , , ,","place":["Chengdu, Sichuan, China, 610064"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Qiushan","family":"Gao","sequence":"additional","affiliation":[{"name":"Sichuan University , , , ,","place":["Chengdu, Sichuan, China, 610064"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yanying","family":"Wang","sequence":"additional","affiliation":[{"name":"Sichuan University , , , ,","place":["Chengdu, Sichuan, China, 610064"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xinyu","family":"Zhang","sequence":"additional","affiliation":[{"name":"Sichuan University , , , ,","place":["Chengdu, Sichuan, China, 610064"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9128-9027","authenticated-orcid":true,"given":"Peng","family":"Wu","sequence":"additional","affiliation":[{"name":"Sichuan University , , , ,","place":["Chengdu, Sichuan, China, 610064"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"316","published-online":{"date-parts":[[2026,7,29]]},"reference":[{"key":"2026081109094266300_cit1","doi-asserted-by":"publisher","DOI":"10.1128\/JCM.01107-21","article-title":"A Real-World Comparison of SARS-CoV-2 Rapid Antigen\nTesting versus PCR Testing in Florida","volume":"59","author":"Allan-Blitz","year":"2021","journal-title":"J. Clin.\nMicrobiol."},{"key":"2026081109094266300_cit2","doi-asserted-by":"publisher","first-page":"126","DOI":"10.1126\/science.abe9187","article-title":"COVID-19 testing:\nOne size does not fit all","volume":"371","author":"Mina","year":"2021","journal-title":"Science"},{"key":"2026081109094266300_cit3","doi-asserted-by":"publisher","DOI":"10.1128\/JCM.01695-20","article-title":"Challenges\nand Controversies to Testing for COVID-19","volume":"58","author":"Binnicker","year":"2020","journal-title":"J.\nClin. Microbiol."},{"key":"2026081109094266300_cit4","doi-asserted-by":"publisher","DOI":"10.1038\/s41467-022-30862-y","article-title":"Rapid, adaptable and sensitive Cas13-based COVID-19\ndiagnostics using ADESSO","volume":"13","author":"Casati","year":"2022","journal-title":"Nat. Commun."},{"key":"2026081109094266300_cit5","doi-asserted-by":"publisher","DOI":"10.1038\/s41467-022-30640-w","article-title":"Combining rapid antigen testing and syndromic surveillance\nimproves\ncommunity-based COVID-19 detection in a low-income country","volume":"13","author":"Chadwick","year":"2022","journal-title":"Nat. Commun."},{"key":"2026081109094266300_cit6","doi-asserted-by":"publisher","DOI":"10.1038\/s41467-022-33460-0","article-title":"Impact of antigen test target failure\nand testing strategies on the transmission of SARS-CoV-2 variants","volume":"13","author":"Del Vecchio","year":"2022","journal-title":"Nat. Commun."},{"key":"2026081109094266300_cit7","doi-asserted-by":"publisher","DOI":"10.1056\/NEJMp2025631","article-title":"Rethinking\nCovid-19 Test Sensitivity\n- A Strategy for Containment","volume":"383","author":"Mina","year":"2020","journal-title":"N. Engl. J. Med."},{"key":"2026081109094266300_cit8","doi-asserted-by":"publisher","first-page":"932","DOI":"10.1038\/s41551-022-00889-z","article-title":"Simplified Cas13-based assays for the fast identification of SARS-CoV-2\nand its variants","volume":"6","author":"Arizti-Sanz","year":"2022","journal-title":"Nat. Biomed. Eng."},{"key":"2026081109094266300_cit9","doi-asserted-by":"publisher","DOI":"10.1038\/s41467-021-21996-6","article-title":"An engineered\nCRISPR-Cas12a variant and DNA-RNA hybrid guides enable robust and\nrapid COVID-19 testing","volume":"12","author":"Ooi","year":"2021","journal-title":"Nat. Commun."},{"key":"2026081109094266300_cit10","doi-asserted-by":"publisher","first-page":"5727","DOI":"10.1021\/acs.analchem.4c01130","article-title":"Integrating Recombinase\nPolymerase Amplification and Photosensitization Colorimetric Detection\nin One Tube for Fast Screening of C. sakazakii in Formula Milk Powder","volume":"96","author":"Wang","year":"2024","journal-title":"Anal. Chem."},{"key":"2026081109094266300_cit11","doi-asserted-by":"publisher","first-page":"1766","DOI":"10.1021\/acs.analchem.5c06890","article-title":"Lateral Flow\nStrip-Compatible\nNucleic Acid Testing for Facile Diagnosis of Infectious Pet Diseases","volume":"98","author":"Zheng","year":"2026","journal-title":"Anal. 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U.S.A."},{"key":"2026081109094266300_cit15","doi-asserted-by":"publisher","first-page":"113","DOI":"10.1016\/S0928-0197(97)00020-2","article-title":"A colorimetric\nPCR-enzyme immunoassay\nto identify hantaviruses","volume":"8","author":"Dekonenko","year":"1997","journal-title":"Clin. Diagn. Virol."},{"key":"2026081109094266300_cit16","doi-asserted-by":"publisher","first-page":"75","DOI":"10.1016\/j.aca.2014.05.032","article-title":"A novel colorimetric\nPCR-based biosensor for detection and quantification of hepatitis\nB virus","volume":"840","author":"Yang","year":"2014","journal-title":"Anal. Chim. Acta"},{"key":"2026081109094266300_cit17","doi-asserted-by":"publisher","first-page":"3141","DOI":"10.1038\/s41596-021-00546-w","article-title":"RNA-extraction-free\nnano-amplified colorimetric test for point-of-care clinical diagnosis\nof COVID-19","volume":"16","author":"Alafeef","year":"2021","journal-title":"Nat. Protoc."},{"key":"2026081109094266300_cit18","doi-asserted-by":"publisher","first-page":"2157","DOI":"10.1002\/anie.201511010","article-title":"A Universal Polymerase\nChain Reaction Developer","volume":"55","author":"Valentini","year":"2016","journal-title":"Angew. Chem., Int.\nEd."},{"key":"2026081109094266300_cit19","doi-asserted-by":"publisher","first-page":"10958","DOI":"10.1021\/ja048749n","article-title":"Label-Free Colorimetric\nDetection of Specific Sequences in Genomic DNA Amplified by the Polymerase\nChain Reaction","volume":"126","author":"Li","year":"2004","journal-title":"J. Am. Chem. Soc."},{"key":"2026081109094266300_cit20","doi-asserted-by":"publisher","first-page":"6456","DOI":"10.1021\/acs.analchem.9b05591","article-title":"Colorimetric\nPolymerase Chain Reaction Enabled by a\nFast Light-Activated Substrate Chromogenic Detection Platform","volume":"92","author":"Dong","year":"2020","journal-title":"Anal. 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Rev."},{"key":"2026081109094266300_cit25","doi-asserted-by":"publisher","first-page":"6624","DOI":"10.1016\/j.tet.2017.10.012","article-title":"A highly selective\ncolorimetric chemosensor for sequential detection of Fe3+ and pyrophosphate\nin aqueous solution","volume":"73","author":"Jang","year":"2017","journal-title":"Tetrahedron"},{"key":"2026081109094266300_cit26","doi-asserted-by":"publisher","first-page":"443","DOI":"10.1016\/j.dyepig.2019.03.058","article-title":"Dual colorimetric\nsensor for picric acid and pyrophosphate:\nPractical application for molecular logic gates","volume":"166","author":"Kumar","year":"2019","journal-title":"Dyes Pigm."},{"key":"2026081109094266300_cit27","doi-asserted-by":"publisher","first-page":"167","DOI":"10.2144\/000113072","article-title":"Colorimetric\nDetection of Loop-Mediated Isothermal Amplification Reaction by Using\nHydroxy Naphthol Blue","volume":"46","author":"Goto","year":"2009","journal-title":"Biotechniques"},{"key":"2026081109094266300_cit28","doi-asserted-by":"publisher","first-page":"293","DOI":"10.1016\/j.bios.2015.08.052","article-title":"Centrifugal\nloop-mediated isothermal\namplification microdevice for rapid, multiplex and colorimetric foodborne\npathogen detection","volume":"75","author":"Oh","year":"2016","journal-title":"Biosens. Bioelectron."},{"key":"2026081109094266300_cit29","doi-asserted-by":"publisher","first-page":"881","DOI":"10.1007\/s00216-016-0084-x","article-title":"Use of a novel metal indicator to judge loop-mediated\nisothermal amplification for detecting the 35S promoter","volume":"409","author":"Wang","year":"2017","journal-title":"Anal. Bioanal. Chem."},{"key":"2026081109094266300_cit30","doi-asserted-by":"publisher","first-page":"1495","DOI":"10.1002\/smll.201302348","article-title":"Ultrasensitive\nand Closed-Tube Colorimetric\nLoop-Mediated Isothermal Amplification Assay Using Carboxyl-Modified\nGold Nanoparticles","volume":"10","author":"Wong","year":"2014","journal-title":"Small"},{"key":"2026081109094266300_cit31","doi-asserted-by":"publisher","DOI":"10.1016\/j.ab.2023.115371","article-title":"Pyrophosphate detection\nmethod using 5-Br-PAPS to detect nucleic acid amplification - Application\nto LAMP method","volume":"684","author":"Hokazono","year":"2024","journal-title":"Anal. Biochem."},{"key":"2026081109094266300_cit32","doi-asserted-by":"publisher","first-page":"298","DOI":"10.1016\/j.cca.2020.10.026","article-title":"A highly sensitive\nand rapid enzymatic method using a biochemical automated analyzer\nto detect inorganic pyrophosphate generated by nucleic acid sequence-based\namplification","volume":"511","author":"Isobe","year":"2020","journal-title":"Clin. Chim. Acta"},{"key":"2026081109094266300_cit33","doi-asserted-by":"publisher","first-page":"1831","DOI":"10.1021\/jacs.4c13768","article-title":"Kinetic and Affinity Profiling Rare\nEarth Metals Using a DNA Aptamer","volume":"147","author":"Wang","year":"2025","journal-title":"J. Am. Chem.\nSoc."},{"key":"2026081109094266300_cit34","doi-asserted-by":"publisher","first-page":"271","DOI":"10.1016\/0890-8508(92)90002-F","article-title":"Amplification of\nan invA gene sequence of Salmonella typhimurium by polymerase chain\nreaction as a specific method of detection of Salmonella","volume":"6","author":"Rahn","year":"1992","journal-title":"Mol. Cell. Probes"},{"key":"2026081109094266300_cit35","doi-asserted-by":"publisher","first-page":"3019","DOI":"10.1016\/j.watres.2009.04.017","article-title":"Analytical comparison\nof nine PCR\nprimer sets designed to detect the presence of Escherichia coli\/Shigella in water samples","volume":"43","author":"Maheux","year":"2009","journal-title":"Water Res."},{"key":"2026081109094266300_cit36","first-page":"119","article-title":"BK virus DNA detection by real-time polymerase chain\nreaction in clinical specimens","volume":"30","author":"Marchetti","year":"2007","journal-title":"New Microbiol."},{"key":"2026081109094266300_cit37","doi-asserted-by":"publisher","first-page":"621","DOI":"10.1002\/ijc.22527","article-title":"Human papillomavirus\ntype distribution\nin invasive cervical cancer and high-grade cervical lesions: A meta-analysis\nupdate","volume":"121","author":"Smith","year":"2007","journal-title":"Int. J. Cancer"},{"key":"2026081109094266300_cit38","doi-asserted-by":"publisher","first-page":"3793","DOI":"10.1002\/jmv.25962","article-title":"Development of a sensitive\nand specific nanoparticle-assisted PCR assay for detecting HPV-16\nand HPV-18 DNA","volume":"92","author":"Ma","year":"2020","journal-title":"J. Med. Virol."},{"key":"2026081109094266300_cit39","doi-asserted-by":"publisher","first-page":"16533","DOI":"10.1021\/acs.analchem.5c02603","article-title":"Visible Light-Driven\nDirect Colorimetric Detection of Nitrite with 3,3\u2032,5,5\u2032-Tetramethylbenzidine","volume":"97","author":"Li","year":"2025","journal-title":"Anal. Chem."}],"container-title":["Analytical Chemistry"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/pubs.acs.org\/doi\/pdf\/10.1021\/acs.analchem.6c01682","content-type":"application\/pdf","content-version":"vor","intended-application":"unspecified"},{"URL":"https:\/\/pubs.acs.org\/ancham\/article-pdf\/98\/31\/22792\/66324117\/acs.analchem.6c01682.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"syndication"},{"URL":"https:\/\/pubs.acs.org\/ancham\/article-pdf\/98\/31\/22792\/66324117\/acs.analchem.6c01682.pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,8,11]],"date-time":"2026-08-11T13:24:51Z","timestamp":1786454691000},"score":1,"resource":{"primary":{"URL":"https:\/\/pubs.acs.org\/ancham\/article\/98\/31\/22792\/5236352\/Colorimetric-PCR-Based-on-Pyrophosphate"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,7,29]]},"references-count":39,"journal-issue":{"issue":"31","published-online":{"date-parts":[[2026,7,29]]},"published-print":{"date-parts":[[2026,8,11]]}},"URL":"https:\/\/doi.org\/10.1021\/acs.analchem.6c01682","relation":{},"ISSN":["0003-2700","1520-6882"],"issn-type":[{"value":"0003-2700","type":"print"},{"value":"1520-6882","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,7,29]]}}}