The finding could lead to the development and application of vector control strategies such as sugar baits, aimed at reducing arbovirus transmission.
The typical symptoms include fever, body ache and conjunctivitis.
With this, the total number down with the virus has gone up to 61 across the State.
The five who tested positive were residents of Anayara, Pettah, Karamana, Poojappura and Killipalam, respectively, in Thiruvananthapuram.
All three are from the Thiruvananthapuram district. Everyone's health is satisfactory and there are no concerns, Health Minister Veena George said.
None of the infected patients are admitted in the hospital and they all are stable.
Efforts to eradicate mosquito breeding grounds in the area have also been intensified.
The most common symptoms of the Zika virus are mild fever and edema (swelling caused by excess fluid trapped in the body's tissues). Some people may also experience redness in the eyes, muscle aches and fatigue.
Till now all the cases of Zika have been reported from Thiruvananthapuram, the state capital.
Earlier in the day, a meeting of the state revenue and health ministers was held on dealing with the Zika virus.