The present disclosure provides a backflow prevention device, comprising: an inlet chamber comprising at least two ports, in which a first port is used for receiving a fluid; a chamber-separating component comprising a separating member and a relief valve; an outlet chamber separated from the inlet chamber by the separating member; the relief valve being operatively connected with the separating member for opening or closing a drain port of the outlet chamber; wherein the fluid is allowed to be delivered from the outlet chamber only when the relief valve is closed by means of a differential pressure applied on the separating member. Also provided is a dialysis apparatus comprising the backflow prevention device. According to the present disclosure, the backflow prevention device has simple structure, reduced dead space and small sizes and can work reliably.