1. Tarbiat Modares University 2. Tarbiat Modares University
Paper based nucleic acid testing (nat) has attracted significant interest in point of care diagnostic field. lateral flow nucleic acid biosensor(lfnab) has the advantages of good biocompatibility, cost effectiveness, simplicity, biodegradability and being portable ,thus lfnabs have been very popular in biomedical, food, agriculture, environmental sciences and so forth. this technique is based on biochemical interaction of probe dna-target dna hybridization.
In this work, we present the development of a chemiluminometric lateral flow biosensor in which, detection is performed by using a conventional digital camera. a sandwich-type hybridization assay was applied using two sequence-specific oligonucleotides. e.coli atcc25922 is chosen as a model, and ribosomal rna was chosen as the sensor target because it exists in high copy numbers in bacterial cells. the detection oligonucleotide probe is biotinylated and streptavidin- horse raddish peroxidase conjugate is used as a reporter. during lateral flow, only 16s rrna target hybridized to detection oligonucleotide were trapped and detected by capture oligonucleotide on test line.
With the assay described here as little as 〖10〗^(5 ) cfu bacteria was detected.
The proposed chemiluminometric lateral flow assay provides a rapid, low cost, and sensitive tool for e.coli detection. it shows great promise for point-of-care diagnosis of urinary tract infection.