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Background: Ethiopia federal ministry of health has been working on increasing access to immunization service by deploying solar refrigerators to primary health care facilities. However, there is limited evidence on cold chain and vaccine management status. Therefore, the objective of this study was to assess knowledge of vaccine handlers and status of cold chain and vaccine management and their associated factors in primary health care facilities of Tigray region Northern Ethiopia. Methods: Institutional based cross-sectional study was conducted in four randomly selected districts of Tigray region, Northern Ethiopia. In each selected district, all primary health care facilities with functional vaccine refrigerators were included in the study. Data were collected using a pre-tested semi-structured questionnaire. The collected data were entered into Epi-data version 3.1 and then exported to Statistical Package for Social Sciences (SPSS) version 21 for analysis. All variables with p-value of < 0.25 in bivariate logistic regression analysis were included in multi-variable model to identify predictors of the dependent variables. Results: In this study, fifty Primary Health Care Facilities (PHCFs) were included with a response rate of 94.4%. The overall level of good knowledge of vaccine handlers and good status of cold chain and vaccine management were 48% (95% CI; 30.7%-62%) and 46% (95%CI; 26.1%-61.3%) respectively. Receiving training on cold chain and vaccine management (AOR = 5.18; 95%CI: 1.48–18.18) was significantly associated with knowledge of vaccine handlers. Furthermore, receiving supportive supervision (AOR = 4.58; 95%CI: 1.04–20.17) and good knowledge of vaccine handlers (AOR = 10.97; 95%CI: 2.67–45.07) were significant associated with cold chain and vaccine management. Conclusions: This study showed that knowledge of vaccine handlers on cold chain and vaccine management was poor. Similarly, the cold chain and vaccine management status was also poor. Therefore, on-site training should be provided to vaccine handlers to increase their knowledge, so as to improve their practices on cold chain and vaccine management. In addition, Programme based supportive supervision is needed to improve cold chain and vaccine management. [ABSTRACT FROM AUTHOR]