Route 27, or Highway 27, may refer to:

• Burke Developmental Road (Queensland)
• Zeehan Highway (Tasmania)

• Alberta Highway 27
• British Columbia Highway 27
• Manitoba Highway 27
• Prince Edward Island Route 27
• Saskatchewan Highway 27

Ontario

• Ontario Highway 27
  • York Regional Road 27, formerly Highway 27 in York Region
  • Simcoe County Road 27, formerly Highway 27 in Simcoe County

• Chile Route 27

• National Route 27

• D27 road (Croatia)

• part of I/27 Highway; Czech: cz:Silnice I/27

• A27 autoroute
• Route nationale 27

• Bundesautobahn 27
• Bundesstraße 27

• A27 motorway, from Kozani to Ptolemaida and from Florina to Niki
• A27 road, a limited-access road from Ptolemaida to Florina
• EO27 road, from Amfissa to Bralos

• National Highway 27 (India)

• Road 27

• N27 road (Ireland)

• Autostrada A27

• Japan National Route 27
• Maizuru-Wakasa Expressway

• Suncheon–Wanju Expressway
• National Route 27

• R-27 regional road (Montenegro)

• A27 motorway (Netherlands)

• New Zealand State Highway 27

• Norwegian County Road 27

• M27 highway (Russia)

• Autovía A-27

• European route E27 (partly also in France and Italy)

• British A27 (Whiteparish-Pevensey)
• M27 (Cadnam-Portsmouth)

• Interstate 27
• U.S. Route 27
• Alabama State Route 27
• Arkansas Highway 27
  • Arkansas Highway 27B
  • Arkansas Highway 27N (former)
• California State Route 27
  • County Route A27 (California)
  • County Route J27 (California)
  • County Route S27 (California)
• Connecticut Route 27
• Florida State Road 27
• Georgia State Route 27
• Idaho State Highway 27
• Illinois Route 27 (former)
• Iowa Highway 27
• K-27 (Kansas highway)
• Louisiana Highway 27
• Maine State Route 27
• Maryland Route 27
  • Maryland Route 27B
  • Maryland Route 27C
  • Maryland Route 27D
  • Maryland Route 27E
• Massachusetts Route 27
• M-27 (Michigan highway)
• Minnesota State Highway 27
  • County Road 27 (Hennepin County, Minnesota)
  • County Road 27 (Ramsey County, Minnesota)
  • County Road 27 (Scott County, Minnesota)
  • County Road 27 (Washington County, Minnesota)
• Mississippi Highway 27
• Missouri Route 27
  • Missouri Route 27 (1922) (former)
• Nebraska Highway 27
  • Nebraska Spur 27D
  • Nebraska Recreation Road 27B
  • Nebraska Recreation Road 27C
• Nevada State Route 27 (former)
• New Hampshire Route 27
• New Jersey Route 27
  • County Route 27 (Bergen County, New Jersey)
  • County Route 27 (Monmouth County, New Jersey)
  • County Route 27 (Ocean County, New Jersey)
• New Mexico State Road 27
• New York State Route 27
  • County Route 27 (Allegany County, New York)
  • County Route 27 (Cattaraugus County, New York)
  • County Route 27 (Clinton County, New York)
  • County Route 27 (Columbia County, New York)
  • County Route 27 (Dutchess County, New York)
  • County Route 27 (Erie County, New York)
  • County Route 27 (Greene County, New York)
  • County Route 27 (Lewis County, New York)
  • County Route 27 (Livingston County, New York)
    • County Route 27B (Livingston County, New York)
  • County Route 27 (Nassau County, New York)
  • County Route 27 (Ontario County, New York)
  • County Route 27 (Oswego County, New York)
  • County Route 27 (Rensselaer County, New York)
  • County Route 27 (Rockland County, New York)
  • County Route 27 (Schoharie County, New York)
  • County Route 27 (Schuyler County, New York)
  • County Route 27 (St. Lawrence County, New York)
  • County Route 27B (Suffolk County, New York) (former)
  • County Route 27 (Ulster County, New York)
  • County Route 27 (Washington County, New York)
  • County Route 27 (Westchester County, New York)
  • County Route 27 (Wyoming County, New York)
  • County Route 27 (Yates County, New York)
• North Carolina Highway 27
• North Dakota Highway 27
• Ohio State Route 27 (1923-1927) (former)
• Oklahoma State Highway 27
• Oregon Route 27
• Pennsylvania Route 27
• South Carolina Highway 27
• South Dakota Highway 27
• Tennessee State Route 27
• Texas State Highway 27
  • Farm to Market Road 27
  • Texas Park Road 27
• Utah State Route 27
• Virginia State Route 27
  • Virginia State Route 27 (1918-1933) (former)
  • Virginia State Route 27 (1940–1953) (former)
• Washington State Route 27
• West Virginia Route 27
• Wisconsin Highway 27

Territories

• Guam Highway 27
• Puerto Rico Highway 27

• List of highways numbered 27A