However, the visitors are only one point behind Steve Clarke's men after they picked up all three points against Israel at the weekend.
Scotland can now boast an impressive record of six wins from their last seven matches across all competitions following their narrow 1-0 victory over Slovakia, with Lyndon Dykes netting the winner for Clarke's side after 54 minutes on the night.
The Tartan Army followed on from their dramatic Euro qualifying playoff penalty shoot-out victory over Israel in perfect fashion and can now open up a four-point gap at the top of Nations League group B2 with a second win on the bounce on Wednesday.
In spite of the fact that Stuart Armstrong tested positive for coronavirus - forcing Clarke to include a number of uncapped players after the midfielder's close contacts Kieran Tierney and Ryan Christie had to self-isolate - Scotland have started as they mean to go on as the Tartan Army eye promotion to League A.
Scotland took the spoils in a 2-1 victory in the reverse fixture back in September, but the Czech Republic may be heading into this bout with a renewed sense confidence following their win at the weekend.
Despite further COVID-19 concerns, Jaroslav Silhavy's side managed to keep the momentum going from their 2-1 friendly victory over Cyprus as they recorded a win by the same scoreline in their Nations League showdown with Israel on Sunday.
The Czech Republic took the lead early through a Joel Abu Hanna own goal before Matej Vydra added a second in the 47th minute, with Eran Zahavi's strike proving nothing more than a mere consolation as Silhavy's men recorded their second victory from three in the 2020-21 Nations League.
The Locomotive are also eyeing promotion to League A and are only one point behind Scotland at this stage, with Wednesday's game bound to prove pivotal in the race for top spot.
The Czech Republic's victory at the weekend - their third from four in all competitions - means that the 1976 European Champions are still yet to draw a fixture since June 2017, and history suggests that run is likely to continue on Wednesday.
Indeed, there has only ever been one stalemate between Scotland and the Czech Republic since the latter team was formed, with the Tartan Army looking to avoid a first defeat against their midweek opponents since 2010.
Scotland Nations League form: DWW
Scotland form (all competitions): WWDWWW
Czech Republic Nations League form: WLW
Czech Republic form (all competitions): WLWLWW
Scotland midfielder John Fleck had to be forced off in the win over Slovakia and is a major doubt for this one, with Callum McGregor and Ryan Jack - both of whom came off the bench on Sunday - vying for a start in midweek.
Clarke sang Andy Considine's praises after the 33-year-old kept a clean sheet on his debut for the Tartan Army, and the Aberdeen stalwart could be in line for a second consecutive start in blue following an injury to Liam Cooper.
As for the visitors, Silhavy may have to contend with another reshuffle similar to that of September amid fresh coronavirus concerns within the squad, with the likes of Filip Novak, Antonin Barak, Adam Hlozek and Borek Dockal all being forced to withdraw.
The Czech Republic have put together a reserve team should the main squad not be able to take to the field in midweek, and it remains to be seen whether Wednesday's game will be a mirror image of last month's meeting between the two sides, where the Czechs had to field a makeshift lineup for the clash with the Tartan Army.
Scotland possible starting lineup:
Marshall; O'Donnell, Considine, Gallagher, Robertson; McTominay; Fraser, McGregor, Jack, McGinn; McBurnie
Czech Republic possible starting lineup:
Vaclik; Kaderabek, Kudela, Hovorka, Boril; Soucek, Kral; Masopust, Darida, Provod; Vydra
We say: Scotland 2-0 Czech Republic
It still remains to be seen whether or not the Czech Republic will be forced to call upon their reserve team for this match, but Scotland did not dominate their opponents' up-and-coming players by any means in September's meeting. Both sides have had little time off to recover from their weekend victories, but with the prospect of the visitors having to name a much-changed lineup, we cannot see any other result than a comfortable win for an in-form Scotland side.