Background: Transfusion-transmissible infections (TTIs) is a critical threat to blood safety. Recent screening combines serology with nucleic acid testing (NAT) to shorten diagnostic window periods and detect occult infection. Objectives: To synthesize recent evidence on laboratory screening strategies in blood donors, serology (HBsAg, anti-HCV, anti-HIV, syphilis) and NAT (HBV DNA, HCV RNA, HIV-1 RNA), and to summarize prevalence, NAT yield, and implementation outcomes. Methods: We followed PRISMA guidance. We included ten original studies that evaluated donor screening outcomes in routine blood-bank settings in Africa, the Middle East, and Asia. Data extracted included setting, assays, sample size, TTI prevalence, and NAT yield or incremental detection. Results: Seroprevalence in included studies varied by region; HBsAg ranged from 2–10% in African settings and 0.5–2% in parts of Asia, with lower anti-HIV and variable anti-HCV rates. Studies implementing ID-NAT, MP-NAT reported additional yield over serology alone, particularly for HBV DNA in HBsAg-negative donations and early HCV infection. Adoption studies also described feasibility at national or tertiary-centre scale. Conclusions: Combining serology with NAT improves detection of window-period and occult infections and enhances blood safety. Programs should tailor panels to local epidemiology, consider anti-HBc policies where HBV is endemic, and evaluate cost effectiveness for national scale-up.