Various approaches for the optical detection of chiral compounds have been developed due to their natural optical activity. Since the advantages of second harmonic generation (SHG) on noble-metal nanoparticles (NPs) have been observed, it would be interesting to study the nonlinear phenomena from chiral compounds attached Ag NPs. In the present work, we fabricated chiral-modified Ag NPs based on the self-assembly process of cysteine and Ag, and carried out the investigation on SHG on modified and unmodified Ag NPs. For modified Ag NPs, either L-Cysteine (L-C) or D-Cysteine (D-C), as a pair of enantiomers, was applied on top of the Ag NPs. The resulting chiral-modified monolayers of L-C/Ag NPs and D-C/Ag NPs exhibit a reversed optical rotation difference (ORD) at linearly ±45° polarization of SH, where no such difference exists for Ag NPs alone. SHG efficiently probes and discriminates L-C from D-C monolayers on the modified Ag NPs, which constitutes a simple and sensitive optical diagnostic of chiral molecules.