A Lateral Flow Assay for the Detection of Leptospira lipL32 Gene Using CRISPR Technology

Area of Science:

• Molecular Biology
• Infectious Disease Diagnostics
• Biotechnology

Background:

• Leptospirosis is frequently misdiagnosed as other febrile illnesses, delaying treatment.
• Accurate, field-deployable diagnostics are crucial for timely leptospirosis management.
• Current quantitative polymerase chain reaction (qPCR) methods are limited by cost and accessibility in rural areas.

Purpose of the Study:

• To develop a precise, low-cost, field-level diagnostic tool for the pathogenic *Leptospira lipL32* gene.
• To overcome the limitations of existing molecular diagnostic techniques for leptospirosis.

Main Methods:

• Development of a CRISPR dFnCas9-based quantitative lateral flow immunoassay.
• Detection of the *Leptospira lipL32* gene using the novel assay.

Main Results:

• The developed assay demonstrated superior performance with a lowest detectable limit of 1 fg/mL.
• The test exhibited high sensitivity and selectivity for *Leptospira* detection.
• Successful application of a low-cost lateral flow device for leptospirosis diagnosis.

Conclusions:

• A novel CRISPR dFnCas9-based assay provides a sensitive and selective method for detecting the *lipL32* gene.
• This low-cost lateral flow immunoassay is suitable for field-level leptospirosis diagnosis.
• The developed diagnostic tool can facilitate prompt therapeutic decisions in resource-limited settings.