An integrated chip capable of performing sample pretreatment and nucleic acid amplification for HIV-1 detection

• Published in: Biosensors & bioelectronics 2013-03, Vol.41, p.484-491
• Main Authors: Wang, Jung-Hao, Cheng, Lie, Wang, Chih-Hung, Ling, Wei-Shuo, Wang, Shainn-Wei, Lee, Gwo-Bin
• Format: Article
• Language: English
• Subjects: Biological and medical sciences; Biosensing Techniques - instrumentation; Biotechnology; Cell Fractionation - instrumentation; DNA, Viral - genetics; DNA, Viral - isolation & purification; Equipment Design; Equipment Failure Analysis; Fundamental and applied biological sciences. Psychology; HIV-1; HIV-1 - genetics; HIV-1 - isolation & purification; Humans; Microfluidic; Microfluidic Analytical Techniques - instrumentation; Nucleic acid amplification; Nucleic Acid Amplification Techniques - instrumentation; Oligonucleotide Array Sequence Analysis - instrumentation; PCR; Reproducibility of Results; Sample pretreatment; Sensitivity and Specificity; Sequence Analysis, DNA - instrumentation; Systems Integration
• ISSN: 0956-5663; 1873-4235
• DOI: 10.1016/j.bios.2012.09.011

This study reports on a microfluidic system equipped with a sample pretreatment device and a nucleic acid amplification device for the rapid diagnosis of the human immunodeficiency virus-1 (HIV-1). The system analyzed proviral deoxyribonucleic acid (DNA) from an HIV-infected Jurkat T cell line. In order to ensure accurate diagnosis among other prevalent B-type strains, simultaneous detections of four conserved HIV-1 B-type DNA fragments were performed in this integrated microfluidic system. The entire protocol including cell lysis, extraction of DNA, polymerase chain reaction (PCR), and optical detection were successfully integrated in order to perform a rapid, automated diagnosis. Experimental results showed that four primer sets with conserved HIV-1 B-type sequences specific for the 167-bp RU5 promoter region, the 424-bp int, the 117-bp tat, and the 162-bp vpr coding regions were successfully amplified from the respective regions of the proviral DNA, even from a single infected cell. This accurate real-time detection was achieved within 95min using the integrated optical system. ► This paper automates diagnosis of HIV-1 proviral DNA. ► Four genes can be detected from the respective regions. ► A new integrated microfluidic system is reported.