Novel Real-Time Fluorescent PCR Chip Applied to Examine the Ankylosing Spondylitis

Abstract

In this article, we introduce a novel real-time polymerase chain reaction (PCR) chip, which integrated the pretreatment of biological sample, the PCR reaction, and the real-time fluorescent detection. We have built a dam in the chamber of the chip and formed a crack underneath it to separate the white blood cell from the whole blood. After the lysis of white blood cells, which were separated from the whole blood, the real-time PCR reaction was produced by the integrated microheaters and resistance temperature detector (RTD), and the real-time fluorescent measurement was made in the identical chamber. Compared with conventional tubular PCR analysis, this chip-based PCR analysis can obtain more accurate results with a smaller amount of samples and reagents. We have designed the corresponding detection and control platform, which is composed of syringe pump module, temperature control module, and fluorescence sampling module. We also used the constructed chip for the examination of HLA-B27 related to ankylosing spondylitis, and the results show that the chip is available for the accurate and rapid analysis of HLA-B27 in whole blood.