Talwars in their petition had alleged that the prosecution had "deliberately" dropped some witnesses whose statements and potential testimonies were in "favour" of the case of the accused.
The special CBI court also rejected another petition of Talwars, seeking a direction to the agency to place on record the disclosure statement made by their compounder Krishna and domestic helps Vijay Mandal and Rajkumar.
"The court today rejected our two applications filed on September 30," said Manoj Sisodia, counsel for Rajesh and Nupur Talwar.
The Talwar couple are standing trial for the murders of their daughter Aarushi and domestic help Hemraj, which took place on the intervening night of May 15-16, 2008.
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So far, seven defence witnesses have been examined in the trial court.
The couple have also filed a petition in Supreme Court seeking direction to the CBI to place before the trial court narco-analysis and brain mapping test reports of their three helpers, who were initially accused in the case.