Forty-eight hours after losing for only the second time in four years in the 100 metres at the Jamaican National Championships and Olympic trials, Bolt had to settle for second place again in the 200m at the National Stadium in Kingston.
Blake chased down his friend and rival in the home straight, usually the part when Bolt pulls clear, to claim the 200m crown in 19.80 seconds, Bolt coming home in 19.83secs. Warren Weir was third in 20.03s.
The 22-year-old now leads this year's world rankings over both distances after clocking 9.75 to win the 100m ahead of a slow-starting Bolt on Saturday.
No longer the best sprinter in his own country, the Olympic champion and world record holder suddenly looks vulnerable just 32 days before the athletics gets under way in London.
Asked if he now has something to prove, he said: "I would say definitely. I am an Olympic champion so I have to show the world I am the best. Next week I come back.
"It's not like I was blown away or anything so I know what I need to get it right. I was very sad with my corner, it was awful, but I guess I haven't been doing a lot of corner work. I have been working more on my 100 metres. But I can't blame it on that."
Blake was, until recently, regarded as a 100m specialist, but last September ran the world's second fastest time, after Bolt, clocking 19.26 in Brussels.
Meanwhile, the British squad for London will be named today with Lynsey Sharp, the Edinburgh AC athlete, hoping for a place in the 800m despite not having achieved the Olympic A standard. She is the form athlete over the distance, winning the UK title at the Olympic Trials in Birmingham and following up last Friday with silver in the European Championships in Helsinki.
The Scots virtually assured of their places are Eilidh Child, Eilish McColgan and Lee McConnell.