{"title":"primspecpcr:用于物种特异性DNA引物设计和特异性测试的Python工具包","authors":"Adam Kuzdraliński","doi":"10.1016/j.softx.2025.102249","DOIUrl":null,"url":null,"abstract":"<div><div>PrimeSpecPCR is an open-source Python toolkit that automates the workflow of species-specific primer design (comprising forward primer, reverse primer, and probe) and validation. The software implements a modular architecture comprising four main components: (1) automated retrieval of genetic sequences from NCBI databases based on taxonomy identifiers; (2) multiple sequence alignment using MAFFT to generate consensus sequences; (3) thermodynamically optimized primer and probe design via Primer3-py; and (4) multi-tiered specificity testing against the NCBI GenBank database. The toolkit features a user-friendly graphical interface and customizable parameters for quantitative PCR (qPCR) applications. PrimeSpecPCR accelerates primer development through parallel processing, automatic caching of intermediate results, and production of interactive HTML reports that visualize specificity profiles across taxonomic groups, while minimizing human errors and ensuring reproducibility of results. This toolkit reduces the time-intensive, labour-demanding processes conventionally required for designing species-specific molecular assays.</div></div>","PeriodicalId":21905,"journal":{"name":"SoftwareX","volume":"31 ","pages":"Article 102249"},"PeriodicalIF":2.4000,"publicationDate":"2025-06-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":"{\"title\":\"PrimeSpecPCR: Python toolkit for species-specific DNA primer design and specificity testing\",\"authors\":\"Adam Kuzdraliński\",\"doi\":\"10.1016/j.softx.2025.102249\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<div><div>PrimeSpecPCR is an open-source Python toolkit that automates the workflow of species-specific primer design (comprising forward primer, reverse primer, and probe) and validation. The software implements a modular architecture comprising four main components: (1) automated retrieval of genetic sequences from NCBI databases based on taxonomy identifiers; (2) multiple sequence alignment using MAFFT to generate consensus sequences; (3) thermodynamically optimized primer and probe design via Primer3-py; and (4) multi-tiered specificity testing against the NCBI GenBank database. The toolkit features a user-friendly graphical interface and customizable parameters for quantitative PCR (qPCR) applications. PrimeSpecPCR accelerates primer development through parallel processing, automatic caching of intermediate results, and production of interactive HTML reports that visualize specificity profiles across taxonomic groups, while minimizing human errors and ensuring reproducibility of results. This toolkit reduces the time-intensive, labour-demanding processes conventionally required for designing species-specific molecular assays.</div></div>\",\"PeriodicalId\":21905,\"journal\":{\"name\":\"SoftwareX\",\"volume\":\"31 \",\"pages\":\"Article 102249\"},\"PeriodicalIF\":2.4000,\"publicationDate\":\"2025-06-28\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"SoftwareX\",\"FirstCategoryId\":\"94\",\"ListUrlMain\":\"https://www.sciencedirect.com/science/article/pii/S235271102500216X\",\"RegionNum\":4,\"RegionCategory\":\"计算机科学\",\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"Q2\",\"JCRName\":\"COMPUTER SCIENCE, SOFTWARE ENGINEERING\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"SoftwareX","FirstCategoryId":"94","ListUrlMain":"https://www.sciencedirect.com/science/article/pii/S235271102500216X","RegionNum":4,"RegionCategory":"计算机科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q2","JCRName":"COMPUTER SCIENCE, SOFTWARE ENGINEERING","Score":null,"Total":0}
PrimeSpecPCR: Python toolkit for species-specific DNA primer design and specificity testing
PrimeSpecPCR is an open-source Python toolkit that automates the workflow of species-specific primer design (comprising forward primer, reverse primer, and probe) and validation. The software implements a modular architecture comprising four main components: (1) automated retrieval of genetic sequences from NCBI databases based on taxonomy identifiers; (2) multiple sequence alignment using MAFFT to generate consensus sequences; (3) thermodynamically optimized primer and probe design via Primer3-py; and (4) multi-tiered specificity testing against the NCBI GenBank database. The toolkit features a user-friendly graphical interface and customizable parameters for quantitative PCR (qPCR) applications. PrimeSpecPCR accelerates primer development through parallel processing, automatic caching of intermediate results, and production of interactive HTML reports that visualize specificity profiles across taxonomic groups, while minimizing human errors and ensuring reproducibility of results. This toolkit reduces the time-intensive, labour-demanding processes conventionally required for designing species-specific molecular assays.
期刊介绍:
SoftwareX aims to acknowledge the impact of software on today''s research practice, and on new scientific discoveries in almost all research domains. SoftwareX also aims to stress the importance of the software developers who are, in part, responsible for this impact. To this end, SoftwareX aims to support publication of research software in such a way that: The software is given a stamp of scientific relevance, and provided with a peer-reviewed recognition of scientific impact; The software developers are given the credits they deserve; The software is citable, allowing traditional metrics of scientific excellence to apply; The academic career paths of software developers are supported rather than hindered; The software is publicly available for inspection, validation, and re-use. Above all, SoftwareX aims to inform researchers about software applications, tools and libraries with a (proven) potential to impact the process of scientific discovery in various domains. The journal is multidisciplinary and accepts submissions from within and across subject domains such as those represented within the broad thematic areas below: Mathematical and Physical Sciences; Environmental Sciences; Medical and Biological Sciences; Humanities, Arts and Social Sciences. Originating from these broad thematic areas, the journal also welcomes submissions of software that works in cross cutting thematic areas, such as citizen science, cybersecurity, digital economy, energy, global resource stewardship, health and wellbeing, etcetera. SoftwareX specifically aims to accept submissions representing domain-independent software that may impact more than one research domain.