A novel surface modification technology for microfluidic disks was developed for multichannel blood-typing detection and irregular antibody screening. The antifouling material, poly(ethylene glycol) methacrylate (PEGMA), was used to modify the surface of the microfluidic disk for improving its hydrophilicity and blood compatibility. With the modification of PEGMA, the hydrophilicity was sufficiently improved with a 44.5% reduction of water contact angle. The modified microfluidic disk also showed good biocompatibility with a reduction of hemolytic index (from 3.4% to 1.2%) and platelet adhesion (from 4.6 × 104/cm2 to 1.9 × 104/cm2). Furthermore, the PEGMA modification technique conducted on the microfluidic disk achieved successful adjustment of burst frequency for each chamber in the microchannel, allowing a sequential addiction of reagents in the test protocol of manual polybrene (MP) blood typing. Clinical studies showed that the proposed MP microfluidic disk method not only performed at extremely high consistency with the traditional tube method in the identification of ABO/RhD blood types, but also accomplished an effective screening method for detecting irregular antibodies. In conclusion, this study demonstrated that the easily mass-produced MP microfluidic disk exhibited good blood-typing sensitivity and was suitable for clinical applications.