Tyrophagus putrescentiae (T. putrescentiae) is a worldwide tored grain pest mite which has do great harm to human’s economy and health. Temperature has a great influence on the population dynamics of T. putrescentiae. In this study, the cDNA of T. putrescentiae HSP70 and HSP90 (designated TpHSP70-1, TpHSP70-2 and TpHSP90) were cloned, and their expression were determined by fluorescent real time quantitative PCR. Results indicate that TpHSP70-1, TpHSP70-2 and TpHSP90 contained an open reading frame of 1,998 bp, 1,984 bp and 2,157 bp that encoded 665, 661 and 718 amino acid residues, respectively. Sequences and phylogenetic analyses suggested that TpHSP70 and TpHSP90 showed high homology with other species. The RT-qPCR results indicated the relative expression level of TpHSP90 and TpHSP70 gene at different stages were different. Also thermal stress could increase its expression significantly. This study indicates that TpHSP70 and TpHSP90 are very important genes for T. putrescentiae to defend against thermal stress.