Abstract
Leptospirosis is an underestimated tropical disease caused by the pathogenic Leptospira species and responsible for several serious health problems. Here, we aimed to develop an ultrasensitive DNA biosensor for the rapid and on-site detection of the Loa22 gene of Leptospira interrogans using a gold nanoparticle–carbon nanofiber composite (AuN/CNF)-based screen-printed electrode. Cyclic voltammetry and electrochemical impedance were performed for electrochemical analysis. The sensitivity of the sensor was 5431.74 μA/cm2/ng with a LOD (detection limit) of 0.0077 ng/μL using cyclic voltammetry. The developed DNA biosensor was found highly specific to the Loa22 gene of L. interrogans, with a storage stability at 4 °C for 180 days and a 6% loss of the initial response. This DNA-based sensor only takes 30 min for rapid detection of the pathogen, with a higher specificity and sensitivity. The promising results obtained suggest the application of the developed sensor as a point of care device for the diagnosis of leptospirosis.
Highlights
Leptospirosis is an acute bacterial septicemic disease caused by the pathogenic spirochaetes of the genus Leptospira [1]
Several methods are available for the detection of leptospirosis that include polymerase chain reaction (PCR), multiplex loopmediated isothermal amplification (m-LAMP), Immunoglobulin-M (IgM)-based enzyme-linked immunosorbent assay (IgM ELISA), and the microscopic agglutination test (MAT) [3,4]
The electrode was characterized at a different phase of the sensor development, including the bare electrode and ssDNA probe-modified electrode, and after hybridization of the DNA sensor (DNA Probe) with single stranded genomic DNA (ssGDNA) of L. interrogans
Summary
Leptospirosis is an acute bacterial septicemic disease caused by the pathogenic spirochaetes of the genus Leptospira [1]. Laboratory-based tests are the most important and effective way to diagnose the disease, as its signs and symptoms are very common and similar to other diseases/disorders. Several methods are available for the detection of leptospirosis that include polymerase chain reaction (PCR), multiplex loopmediated isothermal amplification (m-LAMP), Immunoglobulin-M (IgM)-based enzyme-linked immunosorbent assay (IgM ELISA), and the microscopic agglutination test (MAT) [3,4]. Among all the above-mentioned, the MAT is considered the gold standard test for the detection of Leptospira serovars [5]. These traditional methods are time-consuming, expensive, and not able to lay bare the infection at the early stages
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