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A sample-in- result -out microfluidic system for β-thalassemia diagnostics via direct whole blood PCR-reverse dot
Chao Ye1, Yan Wei2, Yilian Zhao3
1School of Clinical Medicine, Guilin Medical University, Guilin, Guangxi, China.
Objective:
Existing thalassemia detection methods demand high - end labs and have complex procedures, leading to long testing cycles. This study aims to develop a convenient detection method integrated with a microfluidic platform for a sample - to - result process.
Method:
First, optimal conditions for the whole blood direct PCR - reverse dot hybridization (dPCR - RDB) system were explored. Then, its performance was evaluated with clinical samples. Finally, the entire process was integrated into a palm - sized microfluidic chip for "sample - in, result - out" detection.
Result:
A stable dPCR - RDB system was established. Clinical verification on 149 samples showed a 0.1 μl whole - blood minimum detection limit, 100 % specificity, and resistance to high triglyceride and bilirubin levels. It had 100 % positive and negative coincidence rates with traditional methods (kappa = 1). The microfluidic - integrated platform achieved "sample - in, result - out" with 0.5-1 μl blood in 130 min, sans a PCR lab.
Conclusion:
A "sample - in, result - out" microfluidic gene detection platform using whole blood as the template was successfully established.
