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Erimin: A Pipeline to Identify Bacterial Strain Specific Primers

Title: Erimin: A Pipeline to Identify Bacterial Strain Specific Primers
Authors: Margaritis Tsifintaris; Paraskevi Koutra; Pavlos Tsiartas; Panagiotis Repanas; Sotirios Touliopoulos; Grigorios Nelios; Anastasia Anastasiadou; Georgia Tamouridou; Anastasios Nikolaou; Ilias Tsochantaridis
Source: DNA ; Volume 6 ; Issue 1 ; Pages: 11
Publisher Information: Multidisciplinary Digital Publishing Institute
Publication Year: 2026
Collection: MDPI Open Access Publishing
Subject Terms: strain-specific primers; comparative genomics; PCR assay development; Whole-Genome Sequencing (WGS); bioinformatics pipeline
Description: Background/Objectives: Strain-level detection of bacteria is essential for applications such as diagnostics, food safety, and microbial monitoring. While 16S rRNA gene sequencing provides genus- or species-level resolution, it cannot reliably discriminate closely related strains. Whole-genome sequencing (WGS) offers high-resolution strain differentiation but remains impractical for routine detection due to cost and analytical complexity. This study aims to enable the translation of WGS data into accurate and cost-effective strain-specific PCR assays. Methods: We developed Erimin, a modular, shell-based bioinformatics pipeline for the automated identification of strain-specific genomic regions from short-read WGS data. Erimin systematically analyzes all available reference genomes for a given bacterial species in combination with sequencing data from a target strain. The workflow integrates reference-based read alignment, extraction of unmapped reads, de novo assembly, contig filtering and validation, genome annotation, and in silico PCR primer design and specificity evaluation. Results: Erimin was applied to Lactiplantibacillus pentosus whole-genome sequencing data to identify genomic regions specific to strain L33 through comparative analysis against a comprehensive set of reference genome assemblies representing multiple Lactiplantibacillus species. These regions were used for in silico PCR primer design and computational specificity assessment against non-target bacterial genomes, supporting discrimination of closely related strains. Conclusions: Erimin provides a structured computational approach for identifying strain-specific genomic regions from WGS data and for supporting the in silico design of PCR primers. This framework facilitates strain-level discrimination using targeted molecular assays.
Document Type: text
File Description: application/pdf
Language: English
Relation: https://dx.doi.org/10.3390/dna6010011
DOI: 10.3390/dna6010011
Availability: https://doi.org/10.3390/dna6010011
Rights: https://creativecommons.org/licenses/by/4.0/
Accession Number: edsbas.5E1A387C
Database: BASE