An integrated chip capable of performing sample pretreatment and nucleic acid amplification for HIV-1 detection.
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 95 min using the integrated optical system.
Department of Power Mechanical Engineering, National Tsing Hua University, Hsinchu, Taiwan.
Jung-Hao
JH
Wang
Lie
L
Cheng
Chih-Hung
CH
Wang
Wei-Shuo
WS
Ling
Shainn-Wei
SW
Wang
Gwo-Bin
GB
Lee
D015374:Biosensing Techniques / Q000295:instrumentation*; D002458:Cell Fractionation / Q000295:instrumentation*; D004279:DNA, Viral / Q000235:genetics / Q000302:isolation & purification*; D004867:Equipment Design; D019544:Equipment Failure Analysis; D015497:HIV-1 / Q000235:genetics / Q000302:isolation & purification*; D006801:Humans; D046210:Microfluidic Analytical Techniques / Q000295:instrumentation*; D021141:Nucleic Acid Amplification Techniques / Q000295:instrumentation*; D020411:Oligonucleotide Array Sequence Analysis / Q000295:instrumentation*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D017422:Sequence Analysis, DNA / Q000295:instrumentation; D018511:Systems Integration
D004279:DNA, Viral