Customs House is a heritage-listed customs house at 427 Queen Street, Brisbane CBD, Brisbane, Queensland, Australia. It was designed by Charles H McLay and built from 1886 to 1889 at a cost of £38,346 by John Petrie & Son. It was originally used for the collection of customs duty and was opened in 1889 when Queensland was a British colony, replacing the original Customs House located at Petrie Bight. It was added to the Queensland Heritage Register on 7 February 2005.

The collection of custom duties on imported products was particularly important to Queensland where the manufacturing sector was slow to develop. Brisbane was declared a port city in 1846. In 1908, seven years after federation, the building was acquired by the federal government.

Customs House is within reach of the CityCat catamaran ferry service, as well as the Free Loop Bus.

The Brisbane Customs House was erected at the northern end of the Town Reach of the Brisbane River, near Petrie's Bight, between 1886 and 1889 to a design prepared by Charles McLay of the Queensland Colonial Architect's Office.

The 1880s building replaced an earlier and much smaller customs house on the site. The location had been chosen in 1849 following the declaration of Moreton Bay as a port of entry in 1846 and after considerable discussion as to the most suitable location for a customs house. At the time shipping activity was centred on the South Brisbane Reach and the decision to locate a customs house at the northern end of the Town Reach acted as an impetus for the development of wharves along this part of the river. A small building was erected for customs purposes in 1850 and in the following decades became increasingly inadequate as Brisbane emerged as the principal commercial centre and port of Queensland.

In 1884 the Queensland Government decided to construct a new customs house. In March 1886 a design by Charles McLay was selected from many proposals in the Colonial Architect's Office for a new Brisbane Customs House. McLay completed the specification in May 1886 and construction commenced in September that year.

Charles McLay was the major designer under the colonial architect George Connolly and was appointed Chief Draftsman in 1889. His works include the Bundaberg Post Office, probably the Fortitude Post Office and the Lady Norman Wing Brisbane Children's Hospital (with John James Clark), of which the Brisbane Customs House is by far the most ambitious and prominent design.

The contract was let to one of Brisbane's oldest and most respected contractors, John Petrie & Son, who tendered with a price of £37,342. The contract time was 30 months but because of difficulties with the supply of some materials, especially stone for the foundations, and alterations to the original plans, the building took longer than anticipated to complete and it was opened on 2 September 1889. Retaining walls, fencing, a double staircase down to the river and nearby earth closets were also constructed.