PNA-immobilized paper for detection of pcr amplicons

Abstract

DNA detection is very important in diverse fields leading to a continuous increase in the research and development of various DNA sensors. This study focuses on the development of a paper-based DNA sensor by using pyrrolidinyl PNA as a recognition probe. The PNA-immobilized DNA sensor was designed to contain several PNA probes in the same device as a self-check system to increase the accuracy and facilitate the data collection and interpretation. The PNA-immobilized paper was then utilized for the detection of the DNA target, followed by the visualization of the captured target DNA via an enzyme-catalyzed colorimetric reaction. In this work, the developed DNA sensor was applied in the diagnosis of HbE which is a common form of thalassemia, and the identification of Bacillus cereus contamination in rice samples. The results showed that the developed sensor can be successfully used for highly specific detection of the mutated gene responsible for HbE thalassemia with a detection limit of 0.1 ng DNA template. For the detection of B. cereus, the sensor showed very high specificity for B. cereus among several related bacteria species with a low detection limit. Due to the excellent specificity of the pyrrolidinyl PNA probe, the DNA detection can be performed at ambient temperature without the requirement of a sophisticated temperature controlling system.