Padraig Amond struck a dramatic stoppage-time winner as Newport County AFC remained at the top of the Sky Bet League Two table.
County enjoyed the better of the chances at Rodney Parade and had to be patient against a Colchester side who had only lost one of their opening eight league matches this term.
But the Exiles broke down a resolute defensive performance from the U's through a wonder-strike from Twine six minutes into the second half.
Substitute Devani Brown thought he had rescued a late point for the away side when he stabbed home on the rebound, before Amond struck in the first minute of added time to ensure County picked up their fourth successive league victory and stayed at the Sky Bet League Two summit.
Manager Michael Flynn admitted that he may have to tinker with his squad due to the Exiles' busy schedule this term, but it was an enforced change to the starting line-up that he had to make against the U's. Saikou Janneh hobbled off during the 3-0 win against Bradford City on Saturday and was replaced by Padraig Amond.
Amond made an immediate impact following his introduction on the weekend by netting after two minutes, and enjoyed the first opportunity of the evening against Colchester. Tristan Abrahams got the better of defender Tom Eastman and his cross found an unmarked Amond, who nodded wide of Dean Gerken's post.
County were in the ascendency and continued to make life difficult for the visitors, as Gerken was brought into action again two minutes later. Twine's ambitious free-kick from 35 yards smashed against the crossbar and forced Gerken to pack-pedal, before the visitors hooked the ball to safety.
But the home side were nearly caught out down the opposite end when Miles Welch-Hayes intercepted Josh Sheehan's pass and slipped in Courtney Senior down the wing. The winger delivered into the area and Luke Norris went agonisngly close to getting on the end of it, but the cross failed to pick out a player in white. It proved to be the away side's best chance of the first half.
County began the second period in the same vein as the first half and broke the dead-lock six minutes after the restart through a rocket from Twine. The Swindon Town loanee has quickly establed himself as a player who has a knack of scoring from distance, and added another spectacular strike to his goalscoring tally. After drifting into a pocket of space, the midfielder fired a 30-yard effort into the top corner, with Gerken unable to keep it out.
Gerken's opposite number Nick Townsend enjoyed a quiet time in between the sticks as the match entered its final stages, although he had to be alert to deny Callum Harriott's powerful effort, which he tipped around the post.
But Townsend was beaten inside the 89th minute when Brown poked the ball in on the rebound after Kwame Poku saw his effort denied.
It looked as though Colchester had chalked up their fifth draw of the campaign before Amond turned home in stoppage time to earn County maxiumum points.
Newport County AFC: Nick Townsend, Ryan Haynes, Mickey Demetriou, Matty Dolan, Brandon Cooper, Liam Shephard, Joss Labadie (C) (Scot Bennett 85'), Josh Sheehan, Scott Twine, Padraig Amond (Kevin Ellison 90+3'), Tristan Abrahams (Robbie Willmott 90').
Subs not used: Tom King, Ashley Baker, Bradley Webb, Jamie Proctor.