An integrated multiplex one-step RT-PCR and Microarray assay for rapid detection and identification of Influenza A and B Viruses using Lab-on-chip technology
Session III - Patrizia Di Pietro – Poster Only
Title of Contribution: An integrated multiplex one-step RT-PCR and Microarray assay for rapid detection and identification of Influenza A and B Viruses using Lab-on-chip technology
Author(s): Patrizia Di Pietro1, Floriana San Biagio1, Monica Capozzoli1, Ong Kian Leong2, Rosemary Tan2
Affiliation(s): STMicroelectronics, Molecular Diagnostic Business Unit, Microfluidics Division, CCI group, Stradale Primosole 50, 95121 Catania, Italy1; Veredus Laboratories Pte Ltd, 83 Science Park Drive #03-02A, The Curie, Singapore Science Park, Singapore 118 2582
Abstract:
The VereFlu Lab-on-chip assay is a qualitative RT-PCR/microarray-based nucleic acid multiplex test, intended for the simultaneous detection and identification of Influenza A types and subtypes H1, H1N1, H3, H3N2, H7, H9, H9N2 and Influenza B through their HA genes and H5 type and H5N1 subtype through their HA and NA genes.
The VereFlu Assay is developed around the ST In-checkTM Platform based on a silicon chip that integrates all the functions needed to identify given oligonucleotide sequences in a sample, including microfluidic handling, a miniaturized PCR reactor and a custom microarray. The chip is mounted on a 1”x3” plastic slide that provides the necessary mechanical, thermal, electrical and fluidic connections.
In addition to the silicon chip a dedicated set of instruments is provided. The integrated compact high precision Temperature Control System designed to enable random access analysis of up to five independent samples. A portable, customized fluorescent-based Optical Reader analyzes the microarray in a few seconds.
Detailed microarray analysis is performed by ST proprietary software and a simple-to-understand diagnostic report is generated. Alternatively, a more technical report can be generated for expert users.
The oligonucleotide primers and probes have been designed to specifically target unique regions of the haemaggultinin (HA) gene of the selected human influenza subtypes. A set of specific primers for H5N1 subtype neuraminase (NA) gene have been designed as a confirmation for that subtype. The fast turn-around time to allow updates of sequences of novel influenza strains in our system offers a flexible and efficient way to quickly detect emergent strains.
Here we report the analytical performance results of the VereFlu Lab-on-chip assay in terms of sensitivity, specificity and reproducibility using in-vitro transcripts of the Influenza A and B genes.
This fast time-to-answer, portable platform with the capability to simultaneously detect numerous target genes is thus particularly suited for diagnostic applications for the detection, differentiation and identification of human influenza subtypes.
