Scholarlink

Purpose <p>This study aimed to optimize a Fourier Transform Infrared (FT-IR) based protocol using the IR Biotyper<sup>®</sup> system for <i>Leptospira</i> strains and to provide the first spectral insights supporting its feasibility for serogroup and serovar differentiation.</p> Methods <p>Reference <i>Leptospira</i> strains were cultured in Ellinghausen–McCullough–Johnson–Harris medium. A sample preparation workflow was optimized to accommodate liquid cultures, including chemical and thermal inactivation protocols to ensure biosafety. FT-IR spectra were acquired using the IR Biotyper<sup>®</sup> spectrometer and analyzed by Principal Component Analysis (PCA) in the 1300–800&#xa0;cm⁻¹ spectral region. Whole Genome Sequencing (WGS) was used as the reference approach for taxonomic confirmation. Identification and serovar differentiation from genomic data were performed using the jSpecies, KmerFinder, and Pathogenwatch platforms.</p> Results <p>The sample preparation methods tested produced spectra suitable for analysis while ensuring biosafety. Spectral acquisition showed good reproducibility for most serovars across technical and biological replicates. Analysis of WGS performed using the jSpecies platform successfully differentiated the two major human-pathogenic serovars, Icterohaemorrhagiae and Copenhageni, whereas other bioinformatics tools showed limitations. PCA of FT-IR spectra generated by the IR Biotyper<sup>®</sup> demonstrated promising discriminatory potential among the serovars and serogroups evaluated, including closely related serovars such as Icterohaemorrhagiae and Copenhageni and serovars within the Australis and Sejroe serogroups.</p> Conclusion <p>The IR Biotyper offers a rapid FT-IR approach for <i>Leptospira</i> typing. Under the experimental conditions evaluated, the method allowed differentiation of some serovars. Although preliminary, these findings suggest that FT-IR may represent a promising complementary tool for epidemiological surveillance of leptospirosis.</p>

错误：搜索内容不能为空，请输入英文关键词
错误：关键词超出字数限制，请精简
高级检索

Application of FT-IR spectroscopy using the IR Biotyper® for Leptospira: protocol optimization and first spectral insights

  • 期刊论文

  • 出版模式：

    开放获取

  • 发表日期：

    2026年4月11日

查看全文
  • Amanda Maria Jesus Bertani,
  • Elaine Santos Lima,
  • Natalia da Cunha Nobrega,
  • Marcelo Sansone,
  • Juliana Alves Garcia,
  • Daniela Leite,
  • Gisele Oliveira de Souza,
  • Tamires Santos de Arruda,
  • Ricardo Polatto,
  • Eliete Calo Romero,
  • Tania Sueli de Andrade,
  • Karoline Rodrigues Campos,
  • Marlon B. N. Santos,
  • Claudio Tavares Sacchi,
  • Marcos Bryan Heinemann,
  • Monique Ribeiro Tiba Casas,
  • Carlos Henrique Camargo,
  • Roberta Morozetti Blanco

摘要

Purpose

This study aimed to optimize a Fourier Transform Infrared (FT-IR) based protocol using the IR Biotyper® system for Leptospira strains and to provide the first spectral insights supporting its feasibility for serogroup and serovar differentiation.

Methods

Reference Leptospira strains were cultured in Ellinghausen–McCullough–Johnson–Harris medium. A sample preparation workflow was optimized to accommodate liquid cultures, including chemical and thermal inactivation protocols to ensure biosafety. FT-IR spectra were acquired using the IR Biotyper® spectrometer and analyzed by Principal Component Analysis (PCA) in the 1300–800 cm⁻¹ spectral region. Whole Genome Sequencing (WGS) was used as the reference approach for taxonomic confirmation. Identification and serovar differentiation from genomic data were performed using the jSpecies, KmerFinder, and Pathogenwatch platforms.

Results

The sample preparation methods tested produced spectra suitable for analysis while ensuring biosafety. Spectral acquisition showed good reproducibility for most serovars across technical and biological replicates. Analysis of WGS performed using the jSpecies platform successfully differentiated the two major human-pathogenic serovars, Icterohaemorrhagiae and Copenhageni, whereas other bioinformatics tools showed limitations. PCA of FT-IR spectra generated by the IR Biotyper® demonstrated promising discriminatory potential among the serovars and serogroups evaluated, including closely related serovars such as Icterohaemorrhagiae and Copenhageni and serovars within the Australis and Sejroe serogroups.

Conclusion

The IR Biotyper offers a rapid FT-IR approach for Leptospira typing. Under the experimental conditions evaluated, the method allowed differentiation of some serovars. Although preliminary, these findings suggest that FT-IR may represent a promising complementary tool for epidemiological surveillance of leptospirosis.