Pasteurella multocida genome assembly of Indonesian local isolates using long-read Oxford Nanopore Technology sequencing.

Abstract

Background: Pasteurella multocida is a Gram-negative bacterium responsible for various diseases in animals, including hemorrhagic septicemia (HS). Oxford Nanopore Technologies' (ONT) platform streams the sequencing data immediately, allowing for on-the-fly analysis. This capability enables researchers to monitor experiments in real time and make prompt decisions, which is particularly beneficial for clinical diagnostics and outbreak surveillance.

Aim: This study presents the genome assembly of two P. multocida isolates using long-read sequencing technology provided by ONT.

Methods: This method involves passing DNA strands through nanopores and detecting changes in electrical current to determine nucleotide sequences. The long-read capability of ONT technology allows for the sequencing of extensive DNA fragments, which is particularly beneficial for resolving complex genomic regions and achieving high-contiguity genome assemblies.

Results: The study generated high-quality single-contig genomes with sizes of 2.35 and 2.29 Mbp and completeness scores exceeding 99% (BUSCO). These assemblies provide valuable resources for functional analyses, enhancing the understanding of virulence factors and the pathogenesis of HS in Indonesian livestock.

Conclusion: This study successfully assembled the genomes of two P. multocida isolates from Lampung (2,952) and Kupang, Nusa Tenggara Timur, Indonesia, using Oxford Nanopore Technologies' long-read sequencing. The resulting single-contig genomes, measuring 2.35 and 2.29 Mbp, respectively, exhibited high completeness scores (BUSCO: >99%). These high-quality assemblies enhance genomic contiguity, providing valuable resources for functional analyses aimed at understanding the virulence factors and pathogenesis of HS in Indonesian livestock.