Neonatal Group B Streptococcal (GBS) infection remains the leading cause of life-threatening infections in newborns in the United States. A rapid and accurate screening test to identify GBS colonization in laboring women is needed. Current research demonstrates that detection of pathogens by PCR can be a rapid method for clinical diagnosis. However, the tedium of conventional DNA sample preparation prior to PCR dampens the enthusiasm of using PCR at point of care. Our preliminary studies both on the bench and with prototypes demonstrate the potential of a miniaturized and automated PCR-ready, DNA sample preparation device, which can be integrated into a handheld PCR machine. The integrated machine is being designed based on microfluidic technology for rapid and accurate GBS detection, so that persons with minimum or no technical training can perform bed site testing for GBS. In this proposal, the primary focus will be to develop, and miniaturize a sample preparation unit, which will lay the foundation for automation of sample preparation and integration into the PCR machine. We expect that results from the proposed studies will be fundamental in providing a solution to the unmet medical need for the rapid and accurate detection of GBS in delivery room.