【Objective】 To explore the correlation between serological screening of human T-lymphotropic virus antibodies (anti HTLV) and Western blot(WB) confirmatory tests among blood donors, so as to explore the infection status of HTLV Ⅰ/Ⅱ in Guangzhou. 【Methods】 The anti HTLV Ⅰ/Ⅱ enzyme-linked immunosorbent assay(ELISA) kit was used to screen voluntary blood donors from Guangzhou Blood Center from July 2016 to August 2022. WB was used to confirm 395 reactive blood samples by ELISA. The correlation between the S/CO values of anti HTLV Ⅰ/Ⅱ ELISA reagents and the confirmatory test was analyzed using ROC curves. 【Results】 The results showed that 25 out of 395 initially screened reactive blood donor samples were confirmed as HTLV positive by WB, while 16 were uncertain. ROC curve analysis showed a correlation between the S/CO values by ELISA and the confirmatory test results: the S/CO value at the highest Youden index was 3.789, which was the optimal threshold. The S/CO value had a certain correlation with the predicted positive rate of confirmatory results (P<0.05): the larger the S/CO value, the higher the predicted positive value. The overall prevalence of HTLV in Guangzhou is relatively low. 【Conclusion】 The prevalence of HTLV among blood donors in Guangzhou is low.Since the false positive rate of HTLV Ⅰ/Ⅱ antibody by ELISA serological screening is high, the confirmatory testing is particularly important.