Background: Homeless youth living on the streets of large cities and engaging in unprotected sex are at risk of infections with C. trachomatis [CT], N. gonorrhoeae [GC], T. vaginalis [TV], and human papillomavirus [HPV]. The APTIMA transcription mediated amplification [TMA] assays are sensitive and specific for detecting genital infections using various sample types. Methods: In Group A, 296 women signed informed consent for collection of 2 cervical swabs [CS] and a liquid-based Pap [L-Pap] [SurePath] sample. In Group 2, 289 women selfcollected a vaginal swab [VS]. CS and VS were tested for CT and GC using APTIMA Combo 2 [AC2] confirmed with ACT and AGC assays. VS were also tested for TV using analyte specific reagents [ASR] in an APTIMA protocol and confirmed using a research TMA directed against alternate targets. The L-Pap sample and CS were tested for 14 high-risk [HR] HPV genotypes using the APTIMA HPV [AHPV] mRNA test [Gen-Probe] and confirmed using HC2 [Qiagen] or HPV Linear Array [LA] [Roche]. Results: Prevalences were as follows: CT 12% [70/585]; GC 2.6% [15/585]; CT and GC 1.5% [9/585]; TV 13.5% [39/289]; HR-HPV 35.5% [105/296] and HR-HPV and CT were present in 7.1% [21/296]. CT, GC and HR-HPV were present in 2 patients. The most common HPV genotypes in descending order were 16, 51, 52, 56, 18, 31, 39, 58, 33, 45, 35, 59, 68, 66. Many samples contained more than 1 genotype, and the greatest number in a patient was 6. Conclusion: Using APTIMA assays to screen genital infections in this younger, largely asymptomatic population demonstrated a high burden of infection. The use of multiple assays enabled confirmation of results and early treatment. Infections with several organisms and HPV genotypes indicate a public health concern for the sexual health of these homeless individuals.