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

• Published in: Sensors (Basel, Switzerland) Vol. 23; no. 14; p. 6544
• Main Authors: Natarajan, Satheesh, Joseph, Jayaraj, Vinayagamurthy, Balamurugan, Estrela, Pedro
• Format: Journal Article
• Language: English
• Published: Switzerland MDPI AG 20-07-2023; MDPI
• Subjects: Antibodies; Communication; CRISPR; dFnCas9; Disease; Enzymes; Genetic testing; Glycerol; Immunoassay; lateral flow device; Leptospira; Leptospirosis; Proteins; United States > US; dFnCas9; CRISPR; lateral flow device; Leptospira
• ISSN: 1424-8220; 1424-8220
• DOI: 10.3390/s23146544

The clinical manifestation of leptospirosis is often misdiagnosed as other febrile illnesses such as dengue. Therefore, there is an urgent need for a precise diagnostic tool at the field level to detect the pathogenic
gene at the molecular level for prompt therapeutic decisions. Quantitative polymerase chain reaction (qPCR) is widely used as the primary diagnostic tool, but its applicability is limited by high equipment cost and the lack of availability in every hospital, especially in rural areas where leptospirosis mainly occurs. Here, we report the development of a CRISPR dFnCas9-based quantitative lateral flow immunoassay to detect the
gene. The developed assay showed superior performance regarding the lowest detectable limit of 1 fg/mL. The test is highly sensitive and selective, showing that leptospirosis diagnosis can be achieved with a low-cost lateral flow device.