The animated series "Johnny Test" does not feature any identifiable LGBTQ+ characters or themes within its narrative. The show primarily focuses on the adventures of Johnny, his super-genius sisters, and their various inventions and escapades.

Johnny Test is an animated children's series centered on the comedic adventures of a young boy, his super-genius sisters, and a talking dog. The show does not feature any identifiable transsexual characters or explore related themes within its narrative, resulting in no depiction.

The show primarily features female characters like Susan and Mary Test, who are scientific geniuses. Their victories in conflict situations are almost exclusively achieved through the use of advanced inventions and technology, rather than direct physical combat, martial arts, or melee weapon skills against male opponents.

Johnny Test is an original animated series, not an adaptation of pre-existing material. All characters were created for the show, establishing their genders from inception. Therefore, no characters were canonically established as a different gender in prior source material.

Johnny Test (2005–2014) is an original animated series, not an adaptation or reboot of a prior work with pre-established character races. All characters were created for this specific show, meaning there was no prior canonical depiction from which their race could have been swapped.