Abstract: Objective To construct a technical platform for detecting the 968delA mutation of PTEN gene in endometrial cancer, and provide the technical support for clinically analyzing the correlation of PTEN gene 968delA mutation with endometrial cancer. Methods The speific 3'-phosphorothioate-modified primers for the 968delA site of PTEN gene were designed. Using wild-type plasmids and the mutant-type plasmids containing the 968delA site of PTEN gene previously constructed in our laboratory as templates, the high-fi-delity polymerase mediated bidirectional primer extension reaction was performed. Then, the conditions of PCR were optimized by the orthogonal design. The optimized mutation-sensitive molecular switch technology was used to detect the 968delA mutation of PTEN gene in 62 clinical endometrial tissue samples collected from the First Affiliated Hospital of Nanhua University. Results The optimal reaction conditions of PCR for detecting the 968delA site of PTEN gene by the mutation-sensitive molecular switch technology were as follows: 30 cycles, 0.3 μL of DNA template ( 10 ng/μL), 0.5 μL of primers ( 10 pumol/L) and annealing temperature of61C, and its sensitivity was up to 10~3 ng/ pμL. The 968delA mutation of PTEN gene was not found in 62 clinical endometrial cancer tissue and 21 normal endometrial tissue samples. Conclusion The technical platform for detecting the 968delA mutation of PTEN gene in endometrial cancer is successfully constructed, which provides a basis for the application of this technology in the 968delA mutation screening of PTEN gene.