We prove a simple necessary and sufficient condition for a two-bridge knot K(p,q) to be quasipositive, based on the continued fraction expansion of p/q. As an application, coupled with some classification results in contact and symplectic topology, we give a new proof of the fact that smoothly slice two-bridge knots are non-quasipositive. Another proof of this fact using methods within the scope of knot theory is presented in Appendix A, by Stepan Orevkov.