Vivek Verma

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.

A novel approach has been developed for the detection of 56 kDa tissue-specific antigen (TSA) gene of Orientia tsutsugamushi a causative agent of scrub typhus disease. The approach was developed by immobilization of 5′ NH2 labeled ssDNA probe selective to 56 kDa TSA gene, to the surface of AuNPs/CNF modified screen-printed electrode. An electrochemical response was recorded with single stranded genomic DNA (ssDNA) of O. tsutsugamushi isolated from patient sample, using cyclic voltammetry and electrochemical impedance spectroscopy. The electrode surface was characterized by Field-Emission Scanning electron microscope (FE-SEM), Fourier Transform Infrared Spectroscopy (FTIR) and Raman Spectroscopy at each step of fabrication. The DNA biosensor shows optimum response within 50–60 s at room temperature (25 ± 3 °C). The sensor shows higher sensitivity [7849 (µA/cm²)/ng DNA], fast response time (60 s), wider linear range (0.04–2.6 ng) with limit of detection of 0.02 ng/µl of ssDNA sample.

Scrub typhus is a mite-borne, acute febrile illness caused by the bacterium Orientia tsutsugamushi. It is a re-emerging infectious disease of the tsutsugamushi triangle. Scrub typhus is transmitted through bites of contaminated chiggers (larval stage). Diagnosis of scrub typhus is challenging as its symptoms mimic with other acute febrile illnesses. Several methods are effectual for diagnosis of scrub typhus that includes enzyme-linked immunosorbent assay (ELISA), immunofluorescence assay (IFA), immunochromatographic test (ICT), Weil–Felix, polymerase chain reaction (PCR) and loop-mediated isothermal amplification (LAMP). Weil–Felix test was initially used for the diagnosis of scrub typhus in underdeveloped countries but not preferred due to a lack of both specificity and sensitivity. Other immuno-based methods like IFA and ELISA are most outrank for detection of scrub typhus due to their higher sensitivity and specificity, but not vigorous to lay bare the infection at early stages and need the convalescent sampling for verification of positive samples. On another deed, PCR based methods becoming acceptable over era due to its dexterity of early-stage diagnosis with higher specificity and sensitivity but lack its applicability in circumstances of scrub typhus due to the variegated genetic makeup of Orientia tsutsugamushi among its serotypes. The present review focused on various detection methods along with their advantages and disadvantages used in the diagnosis of scrub typhus. A comparison between available methods of diagnosis with challenges in the detection of scrub typhus is also summarized.