This study detected the HIV-1 antibody in gingival crevicular fluid (GCF) using gelatin particles aggregation less-sensitive (PA-LS) method, which aimed to provide a foundation for non-invasive HIV infection test in dental clinic. GCF samples and serum samples were collected from three groups, including HIV-1 positive group, drug users group and general population group. Simultaneously the HIV-1 antibodies in GCF samples were detected by PA-LS, and the HIV-1 antibodies in serum samples were screened by ELISA and confirmed by western blot. According to the final results of western blot, it was evaluated about the sensitivity, specificity, omission diagnostic rate, mistake diagnostic rate, positive predictive value and negative predictive value of PA-LS method. For HIV-1 positive group, the sensitivity of PA-LS used to detect HIV-1 antibody in GCF samples was 100%, the omission diagnostic rate was 0. For drug users group, when used to detect the HIV-1 antibody, the sensitivity, specificity, omission diagnostic rate, mistake diagnostic rate, positive predictive value and negative predictive value of PA-LS were 100%, 99.10%, 0%, 0.90%, 95.57%, and 100%, respectively; the results of ELISA/WB and PA-LS had a high consistency (Kappa >0.8, P<0.01) and no significant difference (P>0.05). For general population, all GCF samples showed HIV-1 antibody negative results. In this study, PA-LS used to test HIV-1 antibody in GCF samples showed a good effect, and was non-invasive, simple, cost-effective compared to ELISA used to detect HIV-1 antibody in serum samples.