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Iflavirus is a group of viruses distributed mainly in arthropod species. We surveyed <i>Tribolium castaneum iflavirus</i> (<i>TcIV</i>) in different laboratory strains and in Sequence Read Archives (SRA) in GenBank. <i>TcIV</i> is highly specific to only <i>T. castaneum</i> and is not found in seven other Tenebrionid species, including the closely related species <i>T. freemani</i>. The same strains from different laboratories and different strains displayed largely different degrees of infections in the examination of 50 different lines by using Taqman-based quantitative PCR. We found that ~63% (27 out of 43 strains) of <i>T. castaneum</i> strains in different laboratories are positive for <i>TcIV</i> PCR with large degrees of variation, in the range of seven orders of magnitude, indicating that the <i>TcIV</i> is highly fluctuating depending on the rearing conditions. The <i>TcIV</i> was prevalent in the nervous system with low levels found in the gonad and gut. The transovarial transmission was supported in the experiment with surface-sterilized eggs. Interestingly, <i>TcIV</i> infection did not show observable pathogenicity. <i>TcIV</i> offers an opportunity to study the interaction between the virus and the immune system of this model beetle species.