Related Experiment Video
Updated: May 24, 2026
Polymerase Chain Reaction and Dot-Blot Hybridization for Leptospira Detection in Water Samples
Published on: June 14, 2024
Detection and quantification of Leptospira interrogans in hamster and rat kidney samples: immunofluorescent imprints
Adenizar D Chagas-Junior1, Caroline L R da Silva, Luciane Marieta Soares
1Gonçalo Moniz Institute, Oswaldo Cruz Foundation, Ministry of Health, Salvador, Bahia, Brazil.
Abstract:
A major limitation in the clinical management and experimental research of leptospirosis is the poor performance of the available methods for the direct detection of leptospires. In this study, we compared real-time PCR (qPCR), targeting the lipL32 gene, with the immunofluorescent imprint method (IM) for the detection and quantification of leptospires in kidney samples from the rat and hamster experimental models of leptospirosis. Using a virulent strain of Leptospira interrogans serovar Copenhageni, a chronic infection was established in the rat model, which were euthanized 28 days post-infection, while the hamster model simulated an acute infection and the hamsters were euthanized eight days after inoculation. Leptospires in the kidney samples were detected using culture isolation, qPCR and the IM, and quantified using qPCR and the IM. In both the acute and chronic infection models, the correlation between quantification by qPCR and the IM was found to be positive and statistically significant (P<0.05). Therefore, this study demonstrates that the IM is a viable alternative for not only the detection but also the quantification of leptospires, particularly when the use of qPCR is not feasible.
Insights
The immunofluorescent imprint method (IM) effectively detects and quantifies leptospires, offering a viable alternative to real-time PCR (qPCR) for leptospirosis research and clinical management.
Area of Science:
- Infectious Diseases
- Microbiology
- Diagnostic Methods
Background:
- Leptospirosis diagnosis is hindered by poor performance of current direct detection methods.
- Accurate detection and quantification of leptospires are crucial for clinical management and research.
Purpose of the Study:
- To compare the efficacy of real-time PCR (qPCR) and the immunofluorescent imprint method (IM) for detecting and quantifying leptospires.
- To evaluate these methods in both acute and chronic experimental leptospirosis models.
Main Methods:
- Kidney samples from infected rats (chronic) and hamsters (acute) were analyzed.
- Leptospira interrogans serovar Copenhageni was used to establish infections.
- Detection and quantification were performed using culture isolation, qPCR targeting the lipL32 gene, and IM.
Main Results:
- A significant positive correlation was observed between qPCR and IM quantification in both acute and chronic infection models (P<0.05).
- IM demonstrated reliable performance for both detection and quantification of leptospires.
Conclusions:
- The immunofluorescent imprint method (IM) is a viable alternative for leptospire detection and quantification.
- IM is particularly useful when real-time PCR (qPCR) is not feasible for leptospirosis diagnosis.
More Related Videos
11:50High-throughput Parallel Sequencing to Measure Fitness of Leptospira interrogans Transposon Insertion Mutants During Golden Syrian Hamster Infection
Published on: December 18, 2017
08:51A Modular Workflow for Quantitative, Structural and Functional Analysis of Leptospira Biofilms
Published on: December 19, 2025