Anime and Manga
- Inverted in Hellsing. Integra has no qualms admitting that the reason why her blood is so delicious is because she's a virgin. Seras more traditionally subverts this trope- she was clearly uncomfortable when Alucard asked her and in all likelihood only answered honestly because she realized that if she answered dishonestly she probably would have been killed on the spot.
- When the Title Drop for Never Been Kissed happens, the other characters assume that this means what it sounds like: that Josie has never had a serious romantic relationship. Josie explains that "never been kissed" is actually a metaphor for a "real" kiss, which she proceeds to define.
- In the horror film The Hazing, the film ends with one surviving male winning the day, with a girl on each arm. The girls joke that he probably never could have even dreamed of such a thing happening, and throughout the film, he's frequently mocked under the assumption that he's a virgin, as well as geeky and weak. After their final mockery of him at the end, he gets fed up and explains to them that he lost his virginity when he was 15 to his (female) baby sitter. She's mentioned twice in the film otherwise, so it's sort of foreshadowing.
- Attempted and failed in The Graduate:
Mrs. Robinson: Benjamin, is this your first time?Ben: Is this - what?Mrs. Robinson: It is, isn't it? It is your first time.Ben: That's a laugh, Mrs. Robinson. That's really a laugh. Ha ha.Mrs. Robinson: You can admit that, can't you?Ben: Are you kidding?Mrs. Robinson: It's nothing to be ashamed of -, etc.
- In Black Swan, when Thomas assumes that Nina's a virgin, she quickly denies it. Whether or not she's telling the truth is left ambiguous, but judging from her Shrinking Violet personality, its probably a lie.
- In Dragon Bones, when Ward is approached by a seductive woman, he recalls how he hasn't been a virgin since his fifteenth year. As he has been Obfuscating Stupidity since he was twelve, and is nineteen at the time, it would be quite logical for him to be a virgin, but apparently he's hot enough to make up for his apparent idiocy. He also found out that he isn't into casual sex because he had some, but other people manage to realize that without having casual sex in the first place, so A Man Is Not a Virgin is actually the most plausible reason for his non-virginity. Or maybe the author felt that there needed to be a good reason for him rejecting the advances of an attractive woman, and a virgin would be too keen on losing his virginity.
- In The Sound and the Fury, Caddy asks her brother Quentin if he's ever had sex. He emphatically replies that he has, but she doesn't believe him.
- In Angel the Fang Gang rescues a wizard's daughter from a virgin sacrifice only to find out that she is not a virgin when she proclaims so in a loud affronted tone.
- In Community, Annie has this when Britta and Shirley assume she's a virgin because Annie has never seen a penis before (it's because her first boyfriend would have sex only in the dark).
- On Red Dwarf, Lister and Rimmer are trapped, presumably to Lister's inevitable death on an ice planet, Lister asks Rimmer to tell the story of how he lost his virginity. When Rimmer hesitates, Lister insinuates that he's a virgin and Rimmer relates the ancedote in short order.
- Mad Men:
Joan: This isn't China. There's no money in virginity.Peggy: I'm not a virgin.(We know Peggy isn't a virgin, but Joan doesn't.)
- Pushing Daisies: Ned was defensive about his "experience" when questioned about it in "Bitches". He claimed, when Emerson and Chuck asked, to have had girlfriends before Chuck, though it's arguable whether any of those relationships were consummated.
- The Doctor in the Doctor Who episode "The Doctor Dances".
Nine hundred years old, me. I've been around a bit. I think you can assume that at some point I've 'danced'.
- Modern Family gets into its third-season episode "Virgin Territory" when Alex hears her father saying "Haley's aversion" as "Haley's a virgin" and laughs, which is how he learns she's not.
- Jay from The Inbetweeners to the point of ridiculousness. He claims to have had sex with more women that he's probably actually met in his entire life. By series 3, these claims get so hilariously ridiculous that they involve copious amounts of Squick.
- Supernatural. In the episode ''Jus in Bello', when they need a virgin sacrifice, Dean assumes that nobody is a virgin.
- Sherlock: Sherlock Holmes's protest of "Sex doesn't alarm me" is answered by his brother Mycroft with "How would you know?"
- A contemporary commentator in the Weimar Republic bemoaned his society’s ‘loose morals’, saying it would be a profound shame for high school girls at the time if it were discovered that they were virgins.
- Herman's Head. Louise is a virgin and gets annoyed when everyone talks of how strange it is.
- Subverted in Bakemonogatari when Senjogahara accuses Koyomi of being a virgin, which he denies. We later learn that they're both virgins.
- Beetle Bailey: In one of the banned strips, Miss Buxley meets a soldier in a bar and they're sitting very close to each other, holding hands and making deep eye contact. The soldier says to her "I'd like to go where no man has gone before.". Miss Buxley then moves in closer to him and says "Too late!".
- In Quest for Glory V, there is a unicorn head mounted on the wall of the Adventurer's Guild. Looking at it produces the line, "They say that unicorns are only visible to virgins. You pretend you can't see it."
- In The Longest Journey, if April asks Flipper Burns how he lost his legs, he wants to know first if she is a virgin. If she says she is, he won't believe her and make up a joke answer to her question, whereas if she says she's not, he'll tell a more plausible tale. It's not clear if April really is a virgin (though her personality would make it likely) but the real issue here is that Burns simply can't believe a pretty girl like April would be one.
- The Archer episode "Swiss Miss" has Anka, a 15-year-old German heiress who's constantly trying to get in Archer's pants, much to his distress. It soon becomes clear she's a bratty virgin who's acting out when she compares revving the throttle on a snowmobile to what she does to guys' "Wilhelms."
Archer: What? No, that's not - oh my God. You've never even seen a Wilhelm!Anka: No! I have seen... dozens of Wilhelms! Scores, even!