In-Check system: A highly integrated silicon Lab-on-Chip for sample preparation, PCR amplification and microarray detection of nucleic acids directly from biological samples

Abstract

The In-Check System is based on a miniaturized silicon lab-on-chip (LoC) where the Polymerase Chain reactor lives together with a customizable microarray module for running a seamless nucleic acid test [1]. This device is designed for accurate temperature performances control, such as accuracy and heating rate provided by both a sophisticated chip calibration process and a precise control by the Temperature Control System (TCS). In addition, the device, is optimized for a microarray fluorescence reading operation by an external instrument, the optical reader (OR). Finally, it is based on microfluidic features that enable to load the chip and fill the reaction chambers without the risk of bubble formation or leaks.In this manuscript are reported the experimental results for the detection of human betaglobine gene (HBB) directly from human cells in less than 2 hours in a silicon reactor. The sample preparation process was entirely performed in one single step into the silicon reactor. It was fully characterized by RT-qPCR. We performed also a comparison study showing higher performances in the LoC silicon reactor than the standard tube. Moreover, the DNA extracted was amplified by PCR, and the resulting product hybridized on the microarray. All the results suggest that the hybridization reactions performed on the silicon LoC can be used to exploit the discriminatory power of microarrays for a specific gene detection.

Miniaturized Electrically Actuated Microfluidic System for Biosensor Applications

“…Since its invention in 1983, polymerase chain reaction (PCR) has been the main method for DNA detection and quantification by using the most recent real-time PCR (qRT-PCR) methodology [8]. Over the last decades, there has been a significantly increase of publications reporting the miniaturized LoCs able to integrate all the functions necessary for acid nucleic analysis starting from the extraction, amplification, and detection either through microarray [4,9] or via qRT-PCR method [10]. In this area, the device miniaturization, the ability to handle small sample volume into micro-nanostructures, and the capability to perform nucleic acid amplification have become indispensable functions for the development of smart portable pointof-care biosensors (PoC).…”

A Point of Care Real Time PCR Platform Based on Silicon Technology

“…Several solutions based on multiple systems to integrate the sample preparation, nucleic acids amplification and detection via real time PCR [6], LAMP [7] or microarray [8] has been reported in the literature. All these miniaturized devices have been designed to enable shorter analysis times, reduce reagent consumption, minimize risk of sample contamination and enhance the assay performance such as sensitivity, selectivity and limit of detection.…”

Extremely integrated device for high sensitive quantitative biosensing

“…Fluorescence based microarrays for DNA sequencing and protein recognition tests are finding widespread application, with several advantages with respect to conventional ELISA tests such as reduced reagent consumption and multiplexing [3][4][5].…”

“…The main drawback of SPAD and SiPM devices is the relatively high dark count rate, often the limiting parameter for the limit of detection (LoD) of a particular experiment, as in the case of chemiluminescence [5,25]. As discussed later in the paper, however, the typical dark count rate is not the limiting aspect in this case due to the peculiarity of the adopted approach.…”