Objective: The manifestations and prognoses of acute-on-chronic liver failure (ACLF) with different precipitating events remain heterogeneous. We aimed to investigate the characteristics and prognosis of patients with hepatotropic viral insult (HVI)-induced hepatitis B-related ACLF (HBV-ACLF). Methods: 452 patients with confirmed diagnosis of ACLF were screened in three medical centers in China, and 203 HBV-ACLF patients with definite acute precipitating events were retrospectively analyzed. According to the precipitating events, HBV-ACLF patients induced by HBV reactivation and super-infection with HAV were classified as the hepatotropic viral insult group and those induced by other factors, as the non-virus insult (NVI) group. The clinical characteristics, predictive scoring model, and prognosis of the two groups were compared. Results: Hepatitis B virus reactivation accounted for the largest proportion (39.9%) among all precipitating events. Exacerbation time frame of the HVI group was significantly longer than that of the NVI group (20 days vs. 10 days, P<0.001). Comparison of intergroup prognosis showed that there was no significant difference in the 28-day mortality (20.9% vs. 13.7%, P=0.125), while the 90-day and 1-year mortality in the HVI group were higher than those in the NVI group (36.3% vs. 24.4%, P=0.014; 39.5% vs. 27.5%, P=0.020, respectively). In the HVI group, the lactic acid-free APASL-ACLF Research Consortium(AARC) had the highest predictive value for 90-day mortality (0.741). Conclusions: The 90-day and 1-year survival rate was lower in HBV-ACLF patients induced by HVI than by NVI. The lactate-free AARC score was a better predictor of short- and long-term prognosis in patients with HVI-induced HBV-ACLF.Trial registration: ChiCTR, ChiCTR1900021539 . Registered 26 February 2019 Retrospectively registered, http://www.chictr.org.cn/showprojen.aspx?proj=36342