Polymerase Chain Reaction and Dot-Blot Hybridization for <i>Leptospira</i> Detection in Water Samples

Eduardo Delgadillo-Tellez; Beatriz Zavaleta-Villa; Daniel Atilano-L&#243;pez; Rigoberto Hern&#225;ndez-Castro; Ang&#233;lica Olivo-D&#237;az; Erika Margarita Carrillo-Casas

doi:10.3791/65435

Journal

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Biology

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Polymerase Chain Reaction and Dot-Blot Hybridization for Leptospira Detection in Water Samples

JoVE Journal
Biology
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JoVE Journal Biology

Polymerase Chain Reaction and Dot-Blot Hybridization for Leptospira Detection in Water Samples

Polymerase Chain Reaction and Dot-Blot Hybridization for Leptospira Detection in Water Samples

983 Views

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06:05 min

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June 14, 2024

DOI:

10.3791/65435-v

DOI:

10.3791/65435-v

•

06:05 min

June 14, 2024

•

16 Views

Eduardo Delgadillo-Tellez¹, Beatriz Zavaleta-Villa¹, Daniel Atilano-López², Rigoberto Hernández-Castro³, Angélica Olivo-Díaz¹, Erika Margarita Carrillo-Casas¹

¹Department of Molecular Biology and Histocompatibility,General Hospital “Dr. Manuel Gea González”, ²Diagnostic Laboratory-Bacteriology, Leptospira Section, Department of Microbiology and Immunology, Veterinary Medicine Faculty,UNAM, ³Department of Ecology of Pathogens,General Hospital “Dr. Manuel Gea González”

Transcript

This protocol is designed to identify the presence of Leptospira DNA in water samples. The dot-blot technique is utilized for this purpose, which enables accurate detection and analysis of the DNA. The dot-blot is an easy and versatile non-isotopic technique and may help to answer key questions about the environmental distribution of Leptospira.

This method has the potential to assess huge water bodies and aid in disease surveillance.

Summary

In this study, a dot-blot application was designed to detect Leptospira from the three main clades in water samples. This method allows for the identification of minimal DNA quantities specifically targeted by a digoxigenin-labeled probe, easily detected by an anti-digoxigenin antibody. This approach is a valuable and satisfactory tool for screening purposes.