Evaluation of Molecular and Nanotechnology technologies for the diagnosis of Dengue viruses causing acute undifferentiated febrile illness

Abstract

Undifferentiated fever is a common presenting symptom in both adults and children in tropical countries Despite the control measures vector borne diseases contribute to majority of the Acute Febrile Illness cases Complex clinical presentation makes it difficult to distinguish one from another based on clinical history and physical examination alone Dengue is the fastest spreading vector borne viral disease It has been reported from 250 countries and endemic in more than 100 countries including India Dengue diagnostics is largely serology based which includes detection of dengue NS1 antigen IgM and IgG Molecular methods like reverse transcriptase PCR RT PCR are a rapid diagnostic tool but available only in the tertiary care centres The available methods are not suitable for field or bedside testing There is a need for the development of indigenous methods that are rapid and cost effective for the detection of dengue viruses as the disease mostly affects people in the lower socio economic strata Advances in nanobiotechnology are promising for the development of rapid viral diagnostics Functionalized nanoparticles ensure rapid detection with high sensitivity and less sophistication The study was conducted in a tertiary care centre after obtaining Institutional Ethics committee approval A total of 350 serum samples were collected during the period of 2014 to 2017 Clinical details were recorded in specific proforma based on WHO 2009 dengue case definition Following viral RNA extraction QIAamp viral RNA Mini kit a real time duplex dengue RTPCR Qiagen One step RTPCR kit was optimised targeting the 3 UTR region Clinical dengue groups based on the WHO2009 case definition probable dengue dengue with warning signs and severe dengue were analyzed in terms of molecular serological and other laboratory co relates A Quantum Dot based fluorescent assay and a Magneto fluorescent immunoassay using iron oxide nanoparticles were developed for detection of dengue virus antigen and evaluated using the real time dengue RT PCR as the go