The DA has argued that the people of Gauteng were denied a voice against e-tolling, because the law was not debated in the provincial legislature.
While the application was dismissed it is significant that both parties will pay their own costs, the DA said in a statement.
"We are studying the judgement and will issue further comment at a later stage," the DA said.
"The fight against e-tolling must continue not just in the courts, but also at the ballot box on 7 May."
The DA approached the court after the Transport Laws and Related Matters Amendment Act was passed into law in September last year.
The amendments allowed for the collection of electronically recorded tolls and the implementation of the electronic toll-collection system.
The DA had argued the amendments were unconstitutional and invalid because they had not been passed according to what it deemed to be proper procedure, which would be with input from the provinces.
The bill was tagged as a section 75 bill - an ordinary bill not affecting provinces - rather than a section 76 bill, which does affect provinces.
The DA's legal team had argued that the amendments to the Sanral and National Roads Act had knock-on effects for provinces.