A Lab-on-a-Chip Device Integrated DNA Extraction and Solid Phase PCR Array for the Genotyping of High-Risk HPV in Clinical Samples.

Cancan Zhu,Guoqing Deng,Ke Yang,Yong Liu,Xinchao Zhu,Junsheng Cui,Ling Zhu,Anzhong Hu

doi:10.3390/mi10080537

Abstract

Point-of-care (POC) molecular diagnostics play a crucial role in the prevention and treatment of infectious diseases. It is necessary to develop portable, easy-to-use, inexpensive and rapid molecular diagnostic tools. In this study, we proposed a lab-on-a-chip device that integrated DNA extraction, solid-phase PCR and genotyping detection. The ingenious design of the pneumatic microvalves enabled the fluid mixing and reagent storage to be organically combined, significantly reducing the size of the chip. The solid oligonucleotide array incorporated into the chip allowed the spatial separation of the primers and minimized undesirable interactions in multiplex amplification. As a proof-of-concept for POC molecular diagnostics on the device, five genotypes of high-risk human papillomavirus (HPV) (HPV16/HPV18/HPV31/HPV33/HPV58) were examined. Positive quality control samples and HPV patient cervical swab specimens were analyzed on the integrated microdevice. The platform was capable of detection approximately 50 copies of HPV virus per reaction during a single step, including DNA extraction, solid-phase PCR and genotype detection, in 1 h from samples being added to the chip. This simple and inexpensive microdevice provided great utility for the screening and monitoring of HPV genotypes. The sample-to-result platform will pave the way for wider application of POC molecular testing in the fields of clinical diagnostics, food safety, and environmental monitoring.

Highlights

Molecular diagnostics can provide genetic information for diagnosis in a way that is fast and convenient
We improved the structure by remodeling the middle valve (V2 and V6) of the pump and hollowing out the third layer of the PDMS, forming a chamber separated from the air
V6 was used as a storage chamber for the reaction solution after the SP-polymerase chain reaction (PCR) finished

Summary

Introduction

Molecular diagnostics can provide genetic information for diagnosis in a way that is fast and convenient. The molecular diagnostics industry, especially for nucleic acid amplification test (NAAT), has been inclined to develop more simple, rapid, specific, and cost-effective platforms for point-of-care (POC) testing [2]. These platforms require integration, automation, the miniaturization of liquid handling, sample processing, analysis to facilitate fast diagnostics, low costs, use by minimally trained personnel, and low contamination risk [3]. A number of studies have been performed to incorporate LOC technology into NAAT to achieve “sample-in answer-out” [4,5,6,7]

Methods

Results

Conclusion