The invention belongs to the technical field of metallurgy, and provides a large-diameter seamless steel tube for high-pressure gas cylinders and a manufacturing method thereof. The large-diameter seamless steel tube for high-pressure gas cylinders comprises the following chemical components in percentage by mass: 0.30-0.60% of C, 0.20-0.50% of Si, 0.20-0.60% of Mn, at most 0.010% of P, at most 0.010% of S, 0.90-1.70% of Cr, 2.50-3.50% of Ni, 0.25-0.55% of Mo, 0.05-0.25% of V and the balance of Fe. The manufacturing method adopts spongy iron and scrap steel as raw materials for steelmaking. The large-diameter seamless steel tube has the advantages of high heat stability, high strength, impact toughness, stable enduring plasticity, high heat resistance, excellent corrosion resistance and excellent fatigue resistance, and the yield strength is not lower than 1000 MPa.