The #99 won three times in Brno in the 250 and 125cc classes but needs his first premier-class victory at the track to put some pressure on the championship leader.
While his Repsol Honda team-mate Andrea Dovizioso secured his first ever premier-class victory at Donington Park, Spaniard Dani Pedrosa had a torrid time. He finally limped home in ninth place and now trails Rossi by 72 points in the championship.
The 125 and 250cc world champion has won support races at Brno and needs a repeat of his Laguna Seca victory to close the gap at the top and also fight off the challengers headed by American Colin Edwards on the Monster Tech 3 Yamaha.
Following his second place at Donington the Texan is just 12 points behind Pedrosa and remains is the leading satellite rider.
The Texan was involved in a tremendous battle with Randy de Puniet at the British Grand Prix but the French LCR rider is struggling to be fit for Brno. He broke his ankle in a Motocross training accident and had a pin fitted in an operation to mend the bone.
Dovizioso arrives brimming with confidence following his superb victory in difficult conditions at Donington to chase his first ever win in Brno, while fellow Italian Marco Melandri will be looking to consolidate his impressive season on the Hayate Kawasaki at circuit where he's won three times in the 125 and 250 cc classes. Melandri is expected to be announced as a 2010 Gresini Honda rider in the near future.
Looking less safe are the likes of Rizla Suzuki riders Chris Vermeulen and Loris Capirossi - whose third place at Brno last year was the most recent GSV-R podium - plus Spaniard Toni Elias, who crashed while challenging for the lead in Donington, but finished second behind Rossi last season in the Czech Republic.
San Carlo Honda Gresini team-mate Alex De Angelis was an impressive fourth in England and also needs more strong results to help his contract negotiations.
One rider assured of the biggest following will be former 125cc World Champion Gabor Talmacsi. Around 25,000 Hungarian fans are expected to make the short trip across the border to support their hero who is beginning to get to grips with the MotoGP class riding the Scot Honda after starting the season in the 250cc class.
Meanwhile, Brno will mark the start of new technical rules which limit each rider to five engines changes during the remaining seven rounds of the season. The rule is designed to limit maintenance and running costs, with each additional engine change punishable by a ten point penalty.