Shortly after her trial ended in August, Letby applied for leave to appeal against her convictions.
She lost the first stage of the process, in which a single judge reviewed her arguments as a paper exercise.
At a two-and-a-half day hearing last month, lawyers for the former nurse renewed efforts to bring an appeal before a panel of three judges.
As the judges have now declined to give the go-ahead for the challenge, this ruling marks the end of the appeal process for Letby.
Dame Victoria previously said it could be reported that her appeal was argued on four points related to the judge at her trial refusing legal applications.
The 34-year-old will face a retrial at Manchester Crown Court in June on a single count that she attempted to murder a baby girl, known as Child K, in February 2016.
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