This is a list of the many characters from the animated television series Johnny Test (including its revival). The main characters are Johnny Test, a fearless 11-year-old boy, and his genetically enhanced dog Dukey, as well as his two sisters Susan and Mary who are inventors.

Main characters Johnny Test

Johnathan Avalon "Johnny" Test is the titular protagonist of the show, the brother of Susan and Mary and the youngest child of Hugh and Lila. He is a mischievous, egotistical, and unpredictable 11-year-old boy who's often called the "kid with the flaming hair" due to his hair having a flame-like appearance. Johnny can live any kid's dream because of his genius sisters, only to find that some aren't worth living. Despite being generally repulsed by girls, he has nurtured attraction towards his classmates Janet and Sissy, the latter out of hostile rivalry. As a procrastinator, Johnny goes to great lengths to avoid any work, often using his sisters' inventions to do so, putting himself and others in danger as a result. He also has a superhero alter ego that he calls "Johnny X", whose superpowers include hurricane hands (speeding wind from his hands), shapeshifting, fire-powered transformation, teleportation, ESP, and "power poots".

Dukey Dukeson Leonard "Dukey" Test (voiced by Louis Chirillo in seasons 1–4, Trevor Devall in seasons 5–8) is Johnny's dog and best friend. He often serves as the voice of reason whenever Johnny is on the verge of making a destructive or hasty decision. Dukey was made anthropomorphic and capable of speech by one of Susan and Mary's inventions; he makes no effort to hide his true nature around Johnny, Mary, and Susan, but reverts to acting like a dog around others, including Hugh and Lila. In public, Dukey poses as a human with very long hair. He was originally named "Poochy" in the 2005 pitch pilot.

Susan and Mary Test Susan and Mary Test (also known as the Test Twins) are Johnny's genius twin sisters and daughters of Hugh and Lila, who frequently use him as a lab rat for various inventions, most of which to impress their neighbor, Gil. Though they generally refuse to help Johnny in his antics, they usually end up doing so anyway due to Johnny blackmailing or manipulating them, or in exchange for Johnny allowing them to use him as a test subject. Their hard-headed demeanor makes them gullible, and they have been tricked by Johnny on various occasions. The Tests attend school at the Porkbelly (formally Mega) Institution of Technology.

Susannah Laura "Susan" Test (voiced by Maryke Hendrikse) is the younger twin. A 13-year-old girl who's the sister of Mary and Johnny and the technical middle child of Hugh and Lila. Susan tends to be more irritable and harsh than Mary is, which often leads to her downfall. In the episode "Johnnymon," she claims to lack sympathy for anyone. Eugene/Bling-Bling expresses fond interest in Susan, much to her disinterest. When Bling-Bling pursues Susan, she tries to bribe Johnny into protecting her. Marionette Louise "Mary" Test (voiced by Brittney Wilson in season 1 and 5, Ashleigh Ball in seasons 2–4 and 6, Emily Tennant in Season 7 and 8) is the elder twin and the oldest child of Hugh and Lila. Mary is more friendly and warm-hearted than Susan. She believes less in science and more in science fiction, which somehow always proves to be correct, despite Susan's scorn. She is also more level-headed and conservative than Susan and speaks her mind much less often.

Secondary characters Hugh Test Hubert "Hugh" Test (voiced by Ian James Corlett) is the grumpy but impractical stay-at-home father of the Test family and husband to Lila. Throughout the series, he is portrayed as the disciplinarian of the Test household since, unlike his wife, he has shown to be the one who is more likely to scold and punish his children, mostly Johnny. Hugh's two biggest obsessions are cleaning and cooking meatloaf which the rest of the Test family and anybody else openly despises. A running gag is that Hugh grounds his children for a month for even the smallest offense or for ridiculous reasons.

Lila Test Lila Test (née Little) (voiced by Kathleen Barr) is the mother of Johnny, Susan, and Mary and wife of Hugh. She has brownish-reddish hair, green eyes (later cyan blue), and wears a typical women's work suit, pearl necklace, and heels. She is a businesswoman who specializes in telemarketing. Due to the demands of her job and the fact she's a workaholic, Lila isn't seen at home as often as the rest of the family, but her family always comes first and she never misses out on vacations and family events. Lila also possesses martial arts skills, was shown to be strong enough to defeat a towering caveman-themed wrestler, and is shown to be less strict than her husband, but sometimes forced against her will to take a break from work to relax. Lila has a scary/crazy side that even Hugh has shown fear when he makes mistakes in front of her. She is a dedicated perfectionist and will have a mental breakdown when pushed too far.

Gil Nexdor Gilbert "Gil" Nexdor (voiced by Andrew Francis) is a handsome 15-year-old boy and Susan and Mary's crush who doesn't even know they exist, despite having lived next door to him since they were born and even having met them face to face on several occasions. He is continuously depicted as intellectually lacking, although his handsome looks compensate for this most of the time. Much to the great ire of Susan and Mary, Gil considers Johnny his friend (whenever Gil appears when Johnny is around, he shouts "Hey, Johnny!"). Due to Susan and Mary's crush, many of their experiments often revolve around him, on some occasions even affecting him directly. However, he can never seem to remember their names save for occasions. His name is actually a parody of the character archetype, Girl next door, of which Gil is a genderbent version of.

Mr. Black and Mr. White Mr. Black (voiced by Bill Mondy) and Mr. White (voiced by Scott McNeil in the original series and Deven Mack in the revival series) are two secret agents who ask the Tests (Johnny, Mary, Susan, and Dukey) for help whenever they cannot handle a crisis alone. They seem to be highly incompetent and somewhat slackers at their job. Ironically, Mr. White is black and Mr. Black is white. Despite first appearing as villains, they're now friends to the Test kids on most occasions. They also have a hidden passion for cooking and dream of a vacation in Fiji.

General The General (voiced by Lee Tockar) is Mr. White and Black's boss and leader of the area 51.1 army base. Loud, slightly incompetent, and forgetful, he frequently takes action whenever the situation becomes too large for the Tests to maintain, although his efforts never fare any better (he even causes the situations sometimes). Even though he doesn't approve of the Test Family and their experiments, he still considers them an ally. The General is prone to overkill and rarely considers what result his actions may have on civilians, to the point he fired at a forest, an inhabited area, and even an amusement park, all in an attempt to destroy a monster. He is also very prone to be a glory hog and gloater when he is successful. The running gags in the series involves the General forgetting something and hesitating to remember and planning to get Johnny, Dukey, Susan and Mary an internship when they graduate. The General also stated in an episode about the test kids “Once you graduate high school, you kids are SO hired!”

Sissy Blakely Elisabeth Olivia "Sissy" Blakely (voiced by Brittney Wilson in Seasons 1 & 5 and Ashleigh Ball in Seasons 2–4 & 6) is a girl who tends to hide her crush on Johnny through bullying and asserting superiority over him. She is 11 years old around the same age as Johnny. Johnny also has a crush on her, but he doesn't seem to be aware of it. The two are constantly competing against each other. If Sissy beats him, she will brag about it. Whenever Johnny wins, he'll either get caught into a situation or brag as much as her. Although seen as his equal, Johnny often cheats most episodes. In "Saturday Night's Alright for Johnny" when Johnny pretends to be nice to her, their feelings are shown (along with Dukey and Missy). She seems to be the victim of the show's twists, adventures, and random outcomes; her misfortune often happens at the end of the episode. In later seasons, they become more friendly to each other and Sissy becomes more aware of her crush on Johnny (admitting to thinking he was cute in the 100th episode). She has blonde hair with a red lightning bolt pattern, multiple ear piercings, and a plaid skirt with pants underneath. She is not present in season 7 and 8.

Bumper Randalls Mitchell "Bumper" Randalls (voiced by Scott McNeil) is a local bully who regularly tortures his peers at Porkbelly Middle School, especially Johnny, as he tortures him more than anyone else. However, he has a sensitive side as he loves roses and even has a rose garden. He has two cats: a white cat called "Cuddles" and an orange tomcat. In addition, Bumper also has a lizard named Mr. Muncher and a dog. Some episodes imply his father wants him to be a wrestler, but he only wants to grow flowers. So Bumper takes his anger out on Johnny, leading to him bullying all the time. In "Johnny X Strikes Back," he gains the power to turn to stone and flight or beat Bumper up. He does not appear in Seasons 7 and 8.

Henry Teacherman Mr. Henry Joseph Teacherman (voiced by Louis Chirillo in Seasons 1–4, Trevor Devall in Season 5–8) is Johnny's teacher who Johnny claims "has it in for him". He seems to think that his students don't try hard enough and has been shown to like it when a student fails a test (rarely giving assignments on topics they never learn). At times, Mr. Teacherman has even conveyed to get sadistic pleasure out of punishing Johnny. Mr. Teacherman is always very tough, but not always to be mean. He believes that deep down, there is a great student inside of everyone including Johnny. It's shown that even Johnny can't reason, trick, or bribe him out of a challenging assignment or decision showing that he can be super stubborn and strict (as in autocratic).

Supporting characters Janet Nelson Jr. Janet Nelson Jr. (voiced by Kathleen Barr) is Johnny's former love interest. Janet was a self-centered popular girl, that was usually mean and inconsiderate to him, appointing him as a loser. She had a love–hate relationship with him, but she is too popular to express the love part. Janet was a regular recurring character in the first season. By the second season, Sissy Blakely took over as Johnny's main love interest leading Janet to be written off. While Janet would continue to make cameos throughout the second and third seasons, by the fourth season, she had no longer been seen. Janet had long black hair parted in the middle with pink barrettes on the side, and wore a pink shirt, white pants, and purple shoes.

Hank Anchorman Hank Anchorman (voiced by James Arnold Taylor) is an anchorman for Porkbelly News. In his first appearance, he had blond hair, was thinner and younger. In later episodes, Hank had brown hair (which was a wig), was older, and had a different face. He often reports about the Test family's shenanigans. He is highly devoted to his job to the point he will risk his well-being for a report/update on a story. Hank is also capable of breaking the fourth wall on certain occasions.

Mayor Howard