Evaluating the efficiency of 24 secondary triage strategies for detecting cervical lesions of self-collected high-risk HPV-positive women

Recently, practice guideline for the early detection of cervical cancer in Korea has been released in this journal [1].It was developed based on preexisting guidelines generated by the American Society for Colposcopy and Cervical Pathology (ASCCP) [2], the National Comprehensive Cancer Network [3], the United States Preventive Services Task Force [4], and the Institute for Clinical Systems Improvement [5].Coincidentally, updated versions of ASCCP guideline [6] and American Cancer Society (ACS) guideline [7] have been issued around the same time.Therefore, the updated contents of the abovementioned guidelines are not reflected in ours.The essential changes of the 2012 ASCCP guidelines from prior 2006 version are as follows [6]: 1) cytology reported as negative but lacking endocervical cells can be managed without any repeat, 2) cytology reported as unsatisfactory requires repeat even if human papillomavirus (HPV) is negative, 3) For atypical squamous cells of undetermined significance (ASC-US) cytology, immediate colposcopy is not an option.The serial cytology option for ASC-US incorporates cytology at 12 months, and then if negative, cytology every 3 years, 4) ASC-US and HPVnegative results should be followed with co-testing at 3 years rather than 5 years, 5) ASC-US and HPV-negative results are insufficient to allow exit from screening at age 65 years, 6) women aged 21-24 years are managed conservatively.Recently reported rates of cytology results reported as negative but lacking endocervical cells have ranged from 10% to 20% [8].Prior guidelines recommended early repeat cytology [9].A recent meta-analysis found that negative cytology had favorable specificity and negative predictive value despite absent endocervical cell component [10].Given that most cytology is performed using liquid-based media, unsatisfactory cytology results arise mainly from insufficient squamous cells [11].The 2012 ASCCP guidelines recommended repeat cytology in 2 to 4 months for women with an unsatisfactory cytology result.Those two issues regarding specimen adequacy were not included in our guidelines.The management of women with ASC-US has also been

Triage tests enhance the efficiency cervical cancer screening based on human papillomavirus (HPV), but the best approach for maximizing programmatic effectiveness is still uncertain, particularly in a real-world scenario. To compare the clinical performance of 6 triage strategies based on liquid-based cytology (LBC) and HPV-16 and HPV-18 genotyping individually or in combination as sequential triage tests to detect cervical intraepithelial neoplasia (CIN) grade 2 or higher among women with high-risk HPV. This diagnostic study of routine cervical cancer screening was conducted at 100 primary health centers in Tlaxcala, Mexico. Women aged 30 to 64 years were recruited from August 1, 2013, to February 24, 2016, as part of the Forwarding Research for Improved Detection and Access for Cervical Cancer Screening and Triage study. Six triage scenarios for referral to colposcopy were examined: (1) LBC testing that found atypical squamous cells of undetermined significance (ASC-US) or worse, (2) positive results in HPV-16 genotyping, (3) positive results in HPV-18 genotyping, (4) positive results in HPV-16/HPV-18 genotyping, (5) positive results in HPV-16 genotyping or, if genotyping results were negative, reflex LBC testing that found ASC-US or worse, and (6) positive results in HPV-16/HPV-18 genotyping or, if genotyping results were negative, reflex LBC testing that found ASC-US or worse. Data were analyzed from October 2017 to August 2018. Liquid-based cytological testing with simultaneous HPV-16 and HPV-18 genotyping. Women whose HPV genotyping results were positive for HPV-16 or HPV-18 or whose LBC results found ASC-US or worse and a random set of negative and normal results were referred to colposcopy with histologic analysis used for disease confirmation. Clinical performance of each test strategy for detection of CIN grade 2 or higher. Secondary outcomes included resource utilization of each triage scenario, measured by the number of tests performed, the referral rate for colposcopy, and the numbers of colposcopies per CIN grade 2 or higher detected. A total of 36 212 women (median [interquartile range] age, 40 [35-47] years) were screened, and 4051 women (11.2%) had high-risk HPV. Of these women, 1109 (24.6%) were found to have HPV-16, HPV-18, or ASC-US or worse. Further histologic testing detected CIN grade 2 or higher in 110 of 788 women (14.0%) who underwent follow-up colposcopy. Sensitivity and specificity for 3 main triage strategies were 42.9% and 74.0% for LBC; 58.3% and 54.4% for HPV-16/HPV-18 genotyping; and 86.6% and 34.0% for HPV-16/HPV-18 genotyping with reflex LBC. The referral rate to colposcopy was 29% for HPV-16/HPV-18 with reflex LBC, which was 2-fold higher than the referral rate of 12% for LBC. Triage of women with high-risk HPV with HPV-16/HPV-18 genotyping with reflex LBC was significantly associated with improvement in detection of CIN grade 2 or higher compared with LBC alone. The benefit of disease prevented may outweigh the cost of increasing requirements for colposcopy services in settings with limited adherence to follow-up after a positive screening result.

The American Society for Colposcopy and Cervical Pathology (ASCCP) National Consensus Conference for the Management of Women With Cervical Cytological Abnormalities and Cervical Cancer Precursors was held on the National Institutes of Health campus in Bethesda, Md, September 6-8, 2001. The conference was attended by 121 representatives from 29 national organizations interested in cervical cancer screening issues. For the first time, guidelines for the management of women with abnormal cervical cytology, developed from evidence-based literature, were presented to delegates from the majority of organizations with interest in cervical cancer screening, voted on, and revised when necessary to achieve a majority two-thirds approval. This development of consensus-approved guidelines is likely to be considered one of the most important milestones to date in the management of women with abnormal cervical cytology. The timing of this Consensus Conference resulted from the convergence of many different factors, including new cytologic terminology developed at the Bethesda 2001 workshop and publication of the enrollment data from the National Cancer Institute's Atypical Squamous Cells of Undetermined Significance (ASC-US)/Low-Grade Squamous Intraepithelial Lesions (LSIL) Triage Study, otherwise known as ALTS. Additionally, new preliminary longitudinal ALTS data provided much of the information on the natural history of abnormal Papanicolaou tests and cervical intraepithelial neoplasia (CIN), as well as data on the performance of both new liquid-based cytology and human papillomavirus (HPV) DNA testing in the management of women following colposcopy. The result was a large database of new information that provided the foundation for the ASCCP Consensus Conference. This article covers only the recommendations of the ASCCP Guidelines that were based in large part on the results of the ALTS trial. Therefore, the focus is on the management of women with equivocal (ASC-US) and low-grade (LSIL) cytologic abnormalities. Management of women with these cytologic abnormalities has been particularly problematic, because individually these women are at least risk for CIN 3 and cancer, yet their sheer numerical dominance ensures that they account for the majority of high-grade CIN detected in the United States in the follow-up of abnormal cervical cytology. Data from ALTS confirmed that women with ASC-US could be safely managed by any of the conventional approaches (repeat Papanicolaou test, immediate colposcopy, or HPV testing), but that the preferred management approach for women having an ASC-US report from liquid-based cytology was to assess the patient's risk by testing for HPV. Additionally, longitudinal ALTS data determined that repeat liquid-based cytology at 6 and 12 months and an HPV test at 12 months were nearly equivalent options in the follow-up of women referred for HPV-positive ASC or LSIL, yet not found to have CIN 2+ at initial colposcopy. Therefore, all follow-up recommendations for women with CIN 1 or lower postcolposcopy findings include these 2 options. The data and the recommendations for the management of ASC-US, ASC cannot exclude high-grade squamous intraepithelial lesion, and LSIL are discussed.

1. Chris L. McGowin, PhD[⇑][1] 1. Louisiana State University Health Sciences Center, Department of Microbiology, Immunology and Parasitology, New Orleans, LA 2. Rodney E. Rohde, PhD, MS, SV, SM(ASCP)CM, MBCM 1. Clinical Laboratory Science Program, College of Health Professions, Texas State University, San Marcos, TX 3. Gerald Redwine, MEd, MT(ASCP) 1. Clinical Laboratory Science Program, College of Health Professions, Texas State University, San Marcos, TX <!-- --> 1. Address for Correspondence: Chris L. McGowin, PhD, Louisiana State University Health Sciences Center, Department of Microbiology, Immunology and Parasitology, 1901 Perdido St.; MEB 6214, New Orleans, LA 70112-2822, 504 568-7281, cmcgow{at}lsuhsc.edu 1. Discuss the historical and epidemiological background of Human Papilloma Virus (HPV) infections. 2. Define current evidence based guidelines for HPV diagnosis and management 3. Justify the epidemiological and clinical rationale for HPV testing in the management of cervical cancer Human Papilloma Virus (HPV) is the most common sexually transmitted infection (STI), and currently is the only vaccine-preventable etiology of urogenital disease. As an STI, HPV is an independent risk factor for virtually all cases of cervical cancer and is associated with anogenital and orolabial warts. Importantly, infection with HPV is a necessary factor in the development of squamous cervical neoplasia despite the fact that most infections and dysplastic abnormalities will not progress to malignant transformation.1-3 Over 100 genotypes of HPV have been identified of which less than 50% are transmitted sexually.4 Of the urogenital HPV types, several have been associated directly with the enhanced risk of cervical cancer.4 In 2012, updated guidelines for cervical cancer screening were put forth by the US Preventative Services Task Force (USPSTF) and the combined partnership of the American Society for Colposcopy and Cervical Pathology (ASCCP), the American Cancer Society (ACS) and the American Society for Clinical Pathology (ASCP). Collectively these guidelines lengthened the time interval between cervical cancer screens and increased the age to begin screening. These evidence-based recommendations indicate the use of either cytology alone or in combination with an FDA-approved HPV test stratified primarily by age, but also by the interval since last screen and hysterectomy status. Compared to cytological investigation alone, co-testing can more informatively direct the need for and method of treating precancerous lesions by more accurately assessing a woman's risk for developing cancer. This article aims to concisely summarize the current guidelines for managing cervical cancer screening, and… ABBREVIATIONS: ACS - American Cancer Society, ASCP - American Society for Clinical Pathology, ASCCP - American Society for Colposcopy and Cervical Pathology, ASCUS - atypical squamous cells of undetermined significance, CIN - cervical intraepithelial neoplasia, FDA - Food and Drug Administration, HPV - Human Papilloma Virus, HR-HPV - high-risk HPV, HSIL - high-grade squamous intraepithelial lesions, LSIL - low-grade squamous intraepithelial lesions, MDx - molecular diagnostics, NAAT - nucleic acid amplification test, NPV - negative predictive value, PPV - positive predictive value, STD - sexually transmitted disease, STI - sexually transmitted infection. 1. Discuss the historical and epidemiological background of Human Papilloma Virus (HPV) infections. 2. Define current evidence based guidelines for HPV diagnosis and management 3. Justify the epidemiological and clinical rationale for HPV testing in the management of cervical cancer [1]: #corresp-1

PurposeTo evaluate the performance of different high-risk human papillomavirus (HR-HPV) genotype models in triaging women with cytological diagnosis of atypical squamous cells of undetermined significance (ASCUS).Patients and MethodsA total of 36,679 Chinese women who underwent cytology and HR-HPV genotyping assessments during cervical cancer screening were enrolled in this study. Women with cytology-proven ASCUS were referred for further screening by colposcopy and biopsy. The study endpoint was histological detection of cervical intraepithelial neoplasia grade 2 or worse (CIN2+) at any of the follow-up visits. The sensitivity, specificity, positive predictive values (PPVs), negative predictive values (NPVs), positive likelihood ratio (PLR) and negative likelihood ratio (NLR) of different HR-HPV genotype combination models were estimated.ResultsIn all, 1675 (4.9%) women were identified as having ASCUS, 1454 women underwent colposcopy and biopsy, and 6.0% (87/1454) women were identified as having CIN2+ lesions. Among those with ASCUS who were identified as having CIN2+, the HR-HPV infection rate was 97.7%, and the prevalence rates of HPV-16, −18, −31, −33, −35, −39, −45, −51, −52, −56, −58, −59, −66 and −68 were 48.3%, 8.0%, 6.9%, 4.6%, 1.1%, 2.3%, 3.4%, 3.4%, 26.4%, 1.1%, 17.2%, 2.3%, 0.0% and 0.0%, respectively. Compared to other HR-HPV-type combination models, the HPV16/18/31/33/52/58 model achieved a higher sensitivity [93.1 (87.8–98.4)], specificity [73.0 (70.7–75.4)], PPV [18.0 (14.5–21.5)], NPV [99.4 (98.9–99.9)], PLR [3.7 (3.1–3.8)] and NLR [0.06 (0.03–0.18)] for the triage of ASCUS patients, but the colposcopy referral rate (30.9%) was significantly lower than that of the recommended HR-HPV model (44.0%).ConclusionThis study confirms that the specific HR-HPV genotype HPV16/18/31/33/52/58 is an alternative strategy for ASCUS triage and can effectively reduce the high burden of colposcopy referrals in China.

Performance of carcinogenic human papillomavirus (HPV) testing and HPV16 or HPV18 genotyping for cervical cancer screening of women aged 25 years and older: a subanalysis of the ATHENA study

This study aimed to determine the prevalence of cervical intraepithelial neoplasia grade 3 or worse (≥CIN3) and cost-effectiveness of human papillomavirus (HPV) genotyping with reflex liquid-based cytology (LBC) for cervical cancer screening in Singapore. Women who were ≥25 years old and undertook co-testing with LBC and HPV-genotyping (Cobas-4800, Roche, USA) for HPV-16, HPV-18 and 12 high-risk HPV types in a single institution were studied retrospectively. A single cervical smear in ThinPrep® PreservCyt® solution (Hologic, USA) was separated for tests in independent cytology and molecular pathology laboratories. The results were reviewed by a designated gynaecologist according to institutional clinical management protocols. Those who tested positive for HPV-16 and/or HPV-18 (regardless of cytology results), cytology showing low-grade squamous intraepithelial lesions (LSIL) or high-grade SIL (HSIL), or atypical squamous cells of undetermined significance (ASCUS) with positive 12 high-risk HPV types were referred for colposcopy. Colposcopy was performed by experienced colposcopists. Cervical biopsy, either directed punch biopsies or excisional biopsy, was determined by a colposcopist. The diagnosis of ≥CIN3 was reviewed by a gynaecologic pathologist. Cost-effectiveness of HPV-based screening in terms of disease and financial burden was analysed using epidemiological, clinical and financial input data from Singapore. Of 1866 women studied, 167 (8.9%) had abnormal cytology (≥ASCUS) and 171 (9.2%) tested positive for high-risk HPV. Twenty-three CIN were detected. Three of the 10 ≥CIN3 cases had negative cytology but positive HPV-16. Compared to cytology, HPV genotyping detected more ≥CIN3 (OR: 1.43). HPV+16/18 genotyping with reflex LBC was superior in terms of cost-effectiveness to LBC with reflex HPV, both for disease detection rate and cost per case of ≥CIN2 detected. Compared to cytology, HPV+16/18 genotyping with reflex LBC detected more ≥CIN3 and was cost-effective for cervical screening in Singapore.

PurposeLittle is known about the performance of extended high-risk human papillomavirus (HR-HPV) genotyping triage of cytology showing atypical squamous cells of undetermined significance (ASC-US). This study aims to evaluate the effectiveness of triage with different HR-HPV genotype models among women with ASC-US.Materials and MethodsIn this study, all women who underwent cervical cytology and HR-HPV genotyping were enrolled from 2014 to 2017 in China, and those with cytology showing ASC-US were referred for colposcopy and/or biopsy. The endpoint was histological detection of cervical intraepithelial neoplasia grade 2 or worse (CIN2+). The outcome indicators were the sensitivity, specificity, positive predictive values (PPVs), negative predictive values (NPVs) and colposcopy referral rates.ResultsIn all, 56,788 women were enrolled in this study, and 2658 (4.97%) women were reported to have ASC-US; 10.1% (242/2393) of women with ASC-US were identified as having CIN2+. The HR-HPV infection rate was 95.0% among all women with ASC-US who were identified as CIN2+, and the top five genotypes with prevalence and risk of CIN2+ were HPV16 (OR=26.38), HPV58 (OR=7.04), HPV18 (OR=4.44), HPV33 (OR=3.38), HPV31 (OR=2.97) and HPV52 (OR=2.96). The HPV16/18/31/33/52/58 model achieved higher sensitivity [91.3 (87.8–94.9)], specificity [70.0 (68.1–72.0)], PPV [25.5 (22.4–28.2)] and NPV [98.6 (97.3–98.7)] for the triage of ASC-US patients than the other HR-HPV-type combination models, but the colposcopy referral rate (36.2%) was significantly lower than that of the recommended HR-HPV nongenotyping model (47.6%).ConclusionThis study confirms that the specific HR-HPV genotype HPV16/18/31/33/52/58 is an alternative strategy for ASC-US triage and can effectively reduce the high burden of colposcopy referrals in China.

EACH YEAR APPROXIMATELY 50 million women undergo Papanicolaou testing in the United States. Of these, approximately 3.5 million (7%) are diagnosed with a cytological abnormality requiring additional follow-up or evaluation. Determining which women with cytological abnormalities are at risk for significant cervical disease, performing appropriate diagnostic workups, and treating cancer precursors present a major public health challenge. There are a number of reasons why comprehensive, evidence-based guidelines are needed for the management of women with cervical cytological abnormalities. One reason is that a National Cancer Institute workshop recently revised the criteria used by cytologists to render certain cytological interpretations, as well as the terminology used for reporting cervical cytology results (ie, the Bethesda System). Other reasons include a better understanding of the pathogenesis and natural history of human papillomavirus (HPV) and cervical cancer precursors, and the availability of data from the National Cancer Institute’s randomized Atypical Squamous Cells of Undetermined Significance/Low-grade Squamous Intraepithelial Lesion (ASCUS/LSIL) Triage Study (ALTS) (D. Solomon, MD, written communication, September 6-8, 2001). Moreover, existing guidelines were developed before sensitive molecular methods for detecting high-risk types of HPV and liquid-based cytology methods became widely available. Data are now available suggesting that these new technologies, when used together, are attractive alternatives to older approaches for managing women with cer-

Chromosomal Biomarkers for Detection of Human Papillomavirus Associated Genomic Instability in Epithelial Cells of Cervical Cytology Specimens

Cervical cancer screening has traditionally been based on cervical cytology. Given the aetiological relationship between human papillomavirus (HPV) infection and cervical carcinogenesis, HPV testing has been proposed as an alternative screening test. To determine the diagnostic accuracy of HPV testing for detecting histologically confirmed cervical intraepithelial neoplasias (CIN) of grade 2 or worse (CIN 2+), including adenocarcinoma in situ, in women participating in primary cervical cancer screening; and how it compares to the accuracy of cytological testing (liquid-based and conventional) at various thresholds. We performed a systematic literature search of articles in MEDLINE and Embase (1992 to November 2015) containing quantitative data and handsearched the reference lists of retrieved articles. We included comparative test accuracy studies if all women received both HPV testing and cervical cytology followed by verification of the disease status with the reference standard, if positive for at least one screening test. The studies had to include women participating in a cervical cancer screening programme who were not being followed up for previous cytological abnormalities. We completed a 2 x 2 table with the number of true positives (TP), false positives (FP), true negatives (TN), and false negatives for each screening test (HPV test and cytology) used in each study. We calculated the absolute and relative sensitivities and the specificities of the tests for the detection of CIN 2+ and CIN 3+ at various thresholds and computed sensitivity (TP/(TP + TN) and specificity (TN/ (TN + FP) for each test separately. Relative sensitivity and specificity of one test compared to another test were defined as sensitivity of test-1 over sensitivity of test-2 and specificity of test-1 over specificity of test-2, respectively. To assess bias in the studies, we used the Quality Assessment of Diagnostic test Accuracy Studies (QUADAS) tool. We used a bivariate random-effects model for computing pooled accuracy estimates. This model takes into account the within- and between-study variability and the intrinsic correlation between sensitivity and specificity. We included a total of 40 studies in the review, with more than 140,000 women aged between 20 and 70 years old. Many studies were at low risk of bias. There were a sufficient number of included studies with adequate methodology to perform the following test comparisons: hybrid capture 2 (HC2) (1 pg/mL threshold) versus conventional cytology (CC) (atypical squamous cells of undetermined significance (ASCUS)+ and low-grade squamous intraepithelial lesions (LSIL)+ thresholds) or liquid-based cytology (LBC) (ASCUS+ and LSIL+ thresholds), other high-risk HPV tests versus conventional cytology (ASCUS+ and LSIL+ thresholds) or LBC (ASCUS+ and LSIL+ thresholds). For CIN 2+, pooled sensitivity estimates for HC2, CC and LBC (ASCUS+) were 89.9%, 62.5% and 72.9%, respectively, and pooled specificity estimates were 89.9%, 96.6%, and 90.3%, respectively. The results did not differ by age of women (less than or greater than 30 years old), or in studies with verification bias. Accuracy of HC2 was, however, greater in European countries compared to other countries. The results for the sensitivity of the tests were heterogeneous ranging from 52% to 94% for LBC, and 61% to 100% for HC2. Overall, the quality of the evidence for the sensitivity of the tests was moderate, and high for the specificity.The relative sensitivity of HC2 versus CC for CIN 2+ was 1.52 (95% CI: 1.24 to 1.86) and the relative specificity 0.94 (95% CI: 0.92 to 0.96), and versus LBC for CIN 2+ was 1.18 (95% CI: 1.10 to 1.26) and the relative specificity 0.96 (95% CI: 0.95 to 0.97). The relative sensitivity of HC2 versus CC for CIN 3+ was 1.46 (95% CI: 1.12 to 1.91) and the relative specificity 0.95 (95% CI: 0.93 to 0.97). The relative sensitivity of HC2 versus LBC for CIN 3+ was 1.17 (95% CI: 1.07 to 1.28) and the relative specificity 0.96 (95% CI: 0.95 to 0.97). Whilst HPV tests are less likely to miss cases of CIN 2+ and CIN 3+, these tests do lead to more unnecessary referrals. However, a negative HPV test is more reassuring than a negative cytological test, as the cytological test has a greater chance of being falsely negative, which could lead to delays in receiving the appropriate treatment. Evidence from prospective longitudinal studies is needed to establish the relative clinical implications of these tests.

Objective: To analyze the epidemiological genotype features of human papillomavirus (HPV) in cervical infection and their risks for cervical precancers among women in Shenzhen area. Methods: A total of 2 717 individuals ranging in age from 30~59 years were recruited in 18 community health centers of Shenzhen city from March 1 to June 15, 2015 by a cluster sampling method. The results of genotype of HPV, liquid-based cytology (LBC), colposcopy and pathology were analyzed. The clinical sensitivity and specificity as well as positive (PPV) and negative (NPV) predictive values of the combination of different HPV genotype in screening the cervical intraepithelial neoplasia (CIN) 2 and above were estimated. Results: The HPV infection rate in Shenzhen area was 15.9% (432/2 717). The most common HPV genotype was HPV52 (22.9%), followed by HPV16 (12.7%), HPV53 (10.0%), HPV51 (8.6%) and HPV58 (8.1%). Compared with HPV16/18 genotyping, HPV33/16 genotyping had a higher sensitivity (57.1% vs. 42.9%, P<0.05) and an analogous specificity (87.3% vs. 86.9%, P>0.05) in predicting CIN2+ . The sensitivity of combination of HPV33/16 genotyping and low grade squamous intraepithelial lesion (LSIL) positive tested by LBC in predicting CIN2+ was 75.0%, significantly higher than 64.3% of atypical squamous cells of undetermined significance (ASC-US) positive tested by LBC alone (P<0.05). The specificities of these two methods mentioned above in predicting CIN2+ were 83.5% and 89.2%, respectively, without statistical difference (P>0.05). Conclusions: Women infected by HPV have distinct risks for CIN2+ according to different high-risk HPV genotypes. The top five risks were HPV 33, 16, 58, 56, and 68. HPV-positive women triaged by LBC LSIL+ combined with HPV33/16 genotyping may be a potential strategy for cervical cancer screening in developed urban area.

Testing for carcinogenic or high-risk human papillomavirus (HPV) DNA has proven utility in cervical cancer screening and in many aspects of clinical management for cervical cancer prevention. However, inappropriate testing increases costs without benefit and potentially results in overtreatment of women. This Statement was developed by The Cytopathology Education and Technology Consortium and has been endorsed by additional professional medical societies as listed below. It is intended as a concise, convenient summary of clinical indications for HPV DNA test utilization based on the American Cancer Society 2002 screening recommendations1 and interim guidance,2 and the 2006 American Society for Colposcopy and Cervical Pathology (ASCCP) consensus management guidelines.3 Circumstances in which HPV DNA testing is considered appropriate, and when such testing is generally not appropriate, are outlined below. This statement and Figure are intended as educational tools and references to improve management of women and reduce inappropriate use of HPV tests.1. High-risk (oncogenic) HPV DNA testing is appropriate in the following circumstances.1.1. Routine cervical cancer screening in conjunction with cervical cytology (dual testing or cotesting) for women 30 years and older1.1.1. For women whose cytology results are negative but are HPV positive, repeat both tests in 12 months (As of March 2009, the US Food and Drug Administration approved a HPV 16/18 genotyping test; per ASCCP guidelines,3 HPV 16- and/or HPV 18–positive women, 30 years and older, are referred directly to colposcopy.)1.1.2. For women whose cytology and HPV results are both negative, repeat both tests only after a 3-year interval1.2. Initial triage management of women 21 years and older with a cytologic result of atypical squamous cells of undetermined significance (ASC-US)1.3. Initial triage management of postmenopausal women with cytologic result of low-grade squamous intraepithelial lesion (LSIL)1.4. Postcolposcopy management of women of any age with initial cytologic result of atypical glandular cells† (AGCs) or atypical squamous cells, cannot exclude a high-grade squamous intraepithelial lesion (ASC-H) (when initial workup does not identify a high-grade lesion)1.5. Postcolposcopy management of women 21 years and older with initial cytologic results of ASC-US or LSIL (when initial colposcopy does not identify a high-grade lesion)1.6. Posttreatment surveillance2. High-risk (oncogenic) HPV DNA testing is generally not appropriate in the following situations;2.1. Routine cervical cancer screening in women younger than 30 years2.2. Routine screening with HPV testing and cervical cytology more often than every 3 years for women 30 years and older whose tests were negative at last screening (see 1.1.2. above)2.3. Initial triage or management of adolescents/young adults (age 20 years and younger) with any abnormal cytologic result; further, if HPV testing is inadvertently performed, the results should not be used to influence patient management2.4. Initial triage of LSIL (except for postmenopausal women; see 1.3 above)2.5. Initial triage of ASC-H, high-grade squamous intraepithelial lesion (HSIL), or AGC†/adenocarcinoma in situ (AIS) in women of any age3. Repeat high-risk (oncogenic) HPV DNA testing should generally not be done before 12 months.3.1. Exceptions include follow-up to atypical glandular cells–not otherwise specified (AGC NOS) when no pathology is found at initial workup and follow-up after treatment for cervical intraepithelial neoplasia (CIN) 2,3; see ASCCP guidelines for specific recommendations on testing intervals34. Testing for low-risk (nononcogenic) HPV types has no role in routine cervical cancer screening or for the evaluation of women with abnormal cervical cytology.Endorsed by the following organizations: ‡ American Cancer Society ‡ American Society for Clinical Pathology ‡ American Society for Colposcopy and Cervical Pathology ‡ American Society of Cytopathology ‡ American Society for Cytotechnology ‡ College of American Pathologists ‡ International Academy of Cytology ‡ Papanicolaou Society of Cytopathology‡ Members of the Cytopathology Education and Technology Consortium.The intent of this summary is to facilitate provider education and to encourage appropriate utilization of HPV testing. Note: Clinical judgment should always be used when applying a guideline to an individual patient because it is impossible to develop guidelines that apply to all situations. Links to the 2006 ASCCP Consensus Guidelines, as well as management algorithms, are available on the ASCCP website at http://www.asccp.org/consensus/cytological.shtml.

Background The detection of cancer-associated types of human papillomavirus (HPV) in cervical specimens predicts the presence and future development of cervical intraepithelial neoplasia (CIN). The purposes of this study were (1) to determine the efficacy of a second-generation assay by hybrid capture (HC II) to detect carcinogenic HPV from residual cervical cells of a liquid-based cervical cytologic specimen, and (2) to compare the performance of this second-generation test with the first-generation hybrid capture (HCT) HPV test of material from direct cervical sampling to detect CIN in women with atypical squamous cells of undetermined significance (ASCUS) and low-grade squamous intraepithelial lesion (LSIL) Papanicolaou (Pap) smear reports. Methods Women with a recent Pap smear report of ASCUS or LSIL had a sampling of the cervix using either an Ayre's spatula and cytobrush or an Accellon device sampling for liquid-based cytologic system HC II HPV testing, followed by a Dacron swab sampling of the cervix for standard HCT HPV testing of the paired specimens. All women received colposcopy examinations including cervical biopsy and endocervical curettage, when indicated, to determine criterion standards for comparison. Results Paired swabs and liquid-based cervical specimens from 242 women were available for testing by standard HCT and the newer HC II HPV DNA assays. The sensitivity, specificity, and positive and negative predictive values for detecting CIN grade 2 or 3 (CIN 2/3) were 61.9%, 57.0%, 12.0%, and 94.0%, respectively, for the HCT test, and 90.5%, 29.4%, 10.9%, and 97.0%, respectively, for the liquid-based cytology HC II assay. When only women with an initial ASCUS Pap smear report were considered, the HC II test results were 88.9%, 40.3%, 9.1%, and 98.2%, respectively. Conclusions Testing for lower genital tract carcinogenic HPV DNA using a cervical cytology liquid transport media residual sample is clinically feasible. The new HC II microplate HPV test achieved a greater test sensitivity for detecting carcinogenic HPV and correspondingly of CIN 2/3 compared with the currently available first-generation HC HPV test. Use of a liquid-based cervical cytology system combined with intermediate triage by HC II testing of residual cells for carcinogenic HPV alone may help to efficiently identify CIN 2/3 in women who have a prior screening Pap smear report of ASCUS.

To assess the utility of makorin ring finger protein 1 (MKRN1) as a marker of cervical pathology.A PROspective specimen collection and retrospective Blinded Evaluation study was conducted. Liquid-based cytology samples were collected from 187 women, embedding all residuals as cell blocks for immunohistochemical staining of MKRN1 and P16 INK4a. Results of liquid-based cervical cytology, immunostained cell block sections, and human papillomavirus (HPV) hybrid capture (with real-time polymerase chain reaction) were analyzed. Clinical outcomes were analyzed overall and in subsets of specimens yielding atypical squamous cells of undetermined significance or low-grade squamous intraepithelial lesions.Makorin ring finger protein 1 positivity and grades (1–3) of cervical intraepithelial neoplasia (CIN) increased in tandem (CIN1, 32.4%; CIN2, 60.0%; and CIN3, 80.0%), reaching 92.3% in invasive cancer. Sensitivity, specificity, positive predictive value, and negative predictive value in detecting CIN2+ via MKRN1 were 73.8%, 76.8%, 75.6%, and 75.0%, respectively. The performance of liquid-based cytology was poorer by comparison (61.3%, 69.5%, 66.2%, and 64.8%, respectively), and HPV assay (versus MKRN1 immunohistochemical staining) displayed lower specificity (67.7%). Combined HPV + MKRN1 testing proved highest in sensitivity, specificity, positive predictive value, and negative predictive value (71.8%, 85.5%, 82.3%, and 76.5%, respectively), whereas corresponding values for cytology + HPV (60.6%, 81.8%, 75.4%, and 69.2%) and cytology + MKRN1 (58.8%, 84.1%, 78.3%, and 67.7%) were all similar. In instances of atypical squamous cells of undetermined significance or low-grade squamous intraepithelial lesions, the HPV + MKRN1 combination performed best by above measures (100%, 72.7%, 73.9%, and 100%), followed by cytology + MKRN1 (100%, 50.0%, 60.7%, and 100%).Makorin ring finger protein 1 displayed greater sensitivity and specificity than liquid-based cytology and proved more specific than HPV assay. In combination testing, MKRN1 + HPV showed the highest sensitivity and specificity levels. The MKRN1 biomarker may be a useful adjunct in primary cervical cytology screening.