Cervical cancer is the second most common cancer in women worldwide, and high-risk human papillomavirus (hrHPV) is the primary causative agent for most cervical cancers and their precursor lesions. Therefore, screening protocols including hrHPV testing have become part of the new cervical cancer screening guidelines developed by the American Cancer Society (ACP), the American Society for Colposcopy and the Cervical (ASCCP), and the American Society for Clinical Pathology (ASCP). The new guidelines state that women ≥30 years of age may be screened with cytology alone and rescreened at 3-year intervals if results are negative; or with combined cytology and hrHPV testing (dual screening) and repeat screening every 5 years if both are negative [1]. More than 150 HPV species have been identified, of which approximately 40 are transmitted through sexual contact and infect the anogenital area. HPV is classified into low-risk and high-risk types according to the risk associated with causing cervical cancer. Low-risk HPV types (lrHPV), which include types 6, 11, 42, 43, 44, 54, 61, 62, 70, 72, and 81, are almost never found in cervical cancer; whereas hrHPV (16, 18, 31, 33, 35, 39, 45, 51, 52, 53, 56, 58, 59, 66, 68, 73, and 82) are seen in cervical, vaginal, vulvar anal and penile cancers. The U.S. Food and Drug Administration (FDA) currently approves five hrHPV tests for clinical use. hrHPV was first approved for the Digene Hybrid Capture 2 High-Risk HPVDNA Test, which detects 13 hrHPV types (16, 18, 31 The Cervista HPV HRand Genfind DNA Extraction Kit tests for hrHPVDNA type 14 (16, 18, 31, 33, 35, 39, 45, 51, 52, 56, 58, 59, 66, 68) and is available to those who are positive. If needed the Cervista HPV16/18 kit can be used to test for HPV16/18 infection. Roche’s Cobas HPVTest detects both HPV16 and HPV18 as well as the other 12 hrHPV types (31, 33, 35, 39, 45, 51, 52, 56, 58, 59, 66, 68). In 2014, the CobasHPV Test received FDA approval and became the first cervical cancer screening method that can be applied alone, without cervical cytology, for women ≥25 years of age. The most recent test to enter the U.S. market is the AptimaHPV Assay, which detects the viral proto-oncogene E6 and E7 mRNAs of hrHPV type 14 (16, 18, 31, 33, 35, 39, 45, 51, 52, 56, 58, 59, 66, 68). According to data from the National Health and Nutrition Examination Survey (2003C2006), the The total HPV prevalence among women aged 14C59 years in the United States was 42.5% (hrHPV prevalence 29%; lrHPV prevalence 28.5%). The prevalence of HPV infection can vary significantly by age, ethnicity, race, poverty level, and number of sexual partners. The most significant difference was age. women aged 14-19 years had the lowest overall HPV positivity rate (32.9%), while women aged 20-24 years had the highest rate (53.8%). A total of 37 HPV types were tested, with lrHPV62 being the most common (6.5%). Among hrHPV, HPV53 was the most common (5.8%), followed by HPV16 (4.7%), HPV51 (4.1%), HPV52 (3.6%), and HPV66 (3.6%). hrHPV18 positivity was only 1.8% [2]. In a study of approximately 1 million U.S. women aged ≥30 years who underwent dual screening, the hrHPV positivity rate was approximately 4% [3]. In women with normal cytology, the prevalence of hrHPVDNA varies widely, partly due to different methods of cytology production (conventional smear or liquid-based production), different methods of hrHPV testing and differences in patient populations as described above, the latter not always taken into account in epidemiological studies. A meta-analysis included 194 studies published worldwide from 1995 to 2009, with an estimated original and corrected hrHPV prevalence of 7.2% and 11.7%, respectively, among 1,016,719 cytologically normal women [4]. In a study by Kaiser Permanente (Portland, OR, USA), 8% of the screened population aged 30 years and older were hrHPV positive and conventional cytology smear negative [5]. Women aged 30 years or older are the population of greatest concern for the new guidelines, with recommendations for dual screening or increased frequency of cytology screening. hrHPV positivity was only 1.9% (490/25,259) among cytology-negative women aged ≥30 years using computer-assisted screening methods, according to UPMC data [6]. In the ATHENA (Addressingthe Need for Advanced HPVDiagnostics) study, the overall prevalence of hrHPV (type 14, Cobas HPVTest) among 32,260 cytologically normal women ≥30 years of age was 6.7%, while HPV16, HPV18 and other type 12 hrHPV had prevalence rates of 1%, 0.5% and 5.2%, respectively [7]. Histological follow-up confirmed that a higher proportion of patients with cytology-negative/hrHPV-positive women had detectable high-grade cervical intraepithelial neoplasia or above (CIN2/3+). In a large study based on clinical data with a mean follow-up of approximately 2 years (23.2 months), CIN2/3+ lesions were detected in 21/849 (2.4%) cytology-negative/hrHPV-positive patients [8]. In the large Portland Kaiser Permanente study, 2% of cytology-negative/hrHPV-positive patients developed CIN3 or more severe lesions after 10 years of follow-up. If all cytology-negative/hrHPV-positive women underwent colposcopy, this could result in a significant increase in the number of colposcopies performed, making colposcopy less efficient. However, long-term observations using non-FDA-approved genotyping studies have found that hrHPV16 or 18-infected individuals have an increased risk of CIN2/3 or more severe lesions than other hrHPV-positive individuals who are not HPV16/18 [3].The 10-year cumulative incidence of CIN3 or more severe lesions in HPV16+ women is 17.2% (95% confidence interval 11.5% to 22.9%) and 13.6% (95% CI = 3.6% to 23.7%) in HPV18+ (HPV16C) women, compared to only 3% (95% CI = 1.5% to 4.2%) in non-HPV16/18 other hrHPV-positive women [5]. In another similar study, the 3-year cumulative risk of finding CIN3 or more severe lesions in cytology-negative/hrHPV-positive patients was 4.7% in hrHPV-positive women using a laboratory-developed (non-FDA-approved) genotyping method. The risk of CIN3 or more severe lesions in non-HPV16 other hrHPV-positive women was 2.4%, rising to 10.6% in HPV16-positive individuals. HPV33-positive and HPV18-positive were also associated with an elevated risk of CIN3 or more severe lesions (both 5.9%) [9]. The new edition of the cervical cancer screening guidelines recommends that cytology-negative women and women aged 30 years and older who are found to be hrHPV-positive on dual screening be monitored using one of two methods: repeat dual screening after 12 months or immediate testing for HPV subtypes (HPV16 alone or testing for HPV16/18). If testing for HPV subtypes immediately, HPV16-positive or HPV16/18-positive women should be referred directly for colposcopy, while HPV16-negative or HPV16/18-negative women should have repeat dual screening after 12 months [1]. The FDA-approved HPV16/18 genotyping test technology was first available for testing with routine gynecologic cervical specimens in 2009. Current consensus guidelines support immediate referral for colposcopy in women ≥30 years of age who are cervical cytology negative but HPV16 or HPV18 positive, as HPV16/18 infection has been shown to increase the risk of high-grade cervical lesions [1]. Although there are some existing retrospective reports of the application of laboratory-developed (non-FDA-approved) genotyping methods to detect HPV typing, there are even fewer reports of their histological follow-up results.The ATHENA trial showed that CobasHPV testing for HPV16 or HPV18 positive women had a higher absolute risk (11.7%) of developing CIN3 or more severe lesions than hrHPV negative women and a relative risk (42%) were significantly higher. Another retrospective UPMC study showed that cytology-negative/hrHPV-positive women ≥30 years of age were detected as HPV16-positive and/or HPV18-positive in 101 (12.3%) of 824 patients using the FDA-approved HPV16/18 genotyping test (Cervista) [10]. hpV16-positive, hpV18-positive, and HPV16/18 simultaneous positivity were detected in 9.1%, 2.4% and 0.7%, respectively. In this group, no cervical cancer was detected at short-term histological follow-up, but high-grade CIN (CIN2/3) was detected in 4/51 (7.8%) cytology-negative/hrHPV-positive (HPV16/18-positive) patients. Despite the short follow-up period (mean 3.5 months) of this study, the histological diagnosis rate of CIN2/3 was 7.8% in cytology-negative/hrHPV-positive (HPV16/18-positive) patients compared with 2.4% in cytology-negative/hrHPV-positive (not genotyped) patients, which was 3-fold higher than the latter. A similar retrospective study with fewer cases included 122 cytology-negative/hrHPV-positive patients tested for HPV16/18 genotyping and subsequent biopsy and found that HPV16/18-positive patients had a higher risk of CIN2/3+ (5.9%) compared to only 2.7% of other hrHPV-positive women with non-HPV16/18 [11]. However, the difference was not statistically significant (P=0.0676). In the clinical management of cytology-negative/hrHPV-positive patients, studies using FDA-approved HPV genotyping assays and longer follow-up are needed to elucidate the validity of the new screening guidelines and the practical value of hrHPV genotyping.