The National League of Cities, representing more than 18,000 communities, was among the groups supporting Mesa's appeal.
Petersen's attorney, David Abney, opposed the appeal. He said the fire department has no history of drug abuse or any drug-related incidents.
Abney said the drug testing program had created "fear and uncertainty." He added, "Simply stated, these firefighters are respected adult professionals entitled to respect and privacy."
Without any comment, the high court rejected the appeal and refused to hear the case.
The justices let stand a ruling that a Catholic charity must include prescription contraceptives in its employee health insurance plan even though church teaching condemns artificial birth control as sinful.
Without comment, the justices rejected an appeal by Catholic Charities of Sacramento of what it called a precedent-setting ruling by the California Supreme Court requiring that contraceptives be included in the prescription drug plan provided to its employees.
The charity said the ruling marked the first time a court has upheld a law forcing a religious institution to pay for conduct that conflicts with its religious beliefs.
California is one of about 20 states that require contraceptive coverage if the health plans have prescription drug benefits. The laws were adopted after lawmakers found that private employee prescription plans without contraceptive benefits discriminated against women.
The California Supreme Court ruled that the charity, incorporated separately from the church, did not qualify as a "religious employer" that was exempt from the law.
It said the charity offered secular services such as counseling, low-income housing and immigration services to people of all faiths, without directly preaching Catholic values.
The Supreme Court declined to hear a challenge to Montana campaign contribution limits by critics who contended the strict rules infringed on free speech rights.
The court, without comment, let stand a 9th U.S. Circuit Court of Appeals decision that the caps on contributions by individuals and political action committees were justified to prevent even the appearance of corruption in the state political process.