P16/Ki-67 Dual Staining in Positive Human Papillomavirus DNA Testing for Predictive Diagnosis of Abnormal Cervical Lesions in Northeastern Thai Women

Objective: Cervical cancer screening can effectively reduce new cervical cancer cases, including in Thailand. The abnormal results are subsequently referred for colposcopy. To avoid unnecessary colposcopy, an efficient triage is still needed for validation. This study aimed to investigate the overall positivity of cytology-based screening, HPV detection, and p16/Ki-67 dual staining and evaluate different triage strategies for predictive diagnosis of abnormal cervical lesions in northeastern Thailand. Methods: Cervical cells were collected from 191 women who came for cervical screening in the gynecological outpatient department during March 2019-February 2020. Pap smear samples were classified into 6 groups including 17 atypical glandular cells (AGC), 21 atypical squamous cells of undetermined significance (ASC-US), 7 atypical squamous cells - cannot exclude HSIL (ASC-H), 26 low-grade squamous intraepithelial lesions (LSILs), 19 high-grade SILs (HSILs) and 101 no squamous intraepithelial lesion (noSIL). Polymerase chain reaction (PCR) was performed for HPV DNA detection. HPV genotyping was determined by reverse line blot hybridization. P16/Ki-67 dual staining was performed by using CINtec PLUS Cytology kit. Biopsies from abnormal screening were collected for surgical pathology classification. Results: High-risk HPV (HR-HPV) infection was 2.97%, 29.41%, 38.10%, 57.14%, 46.15% and 84.21% in noSIL, AGC, ASC-US, ASC-H, LSIL and HSIL cytology respectively. P16/ Ki-67 in noSIL, AGC, ASC-US, ASC-H, LSIL and HSIL was 0.99%, 5.88%, 9.52%, 42.86%, 26.92% and 63.16%, respectively (P-value < 0.001). Among p16/Ki-67 positive cases, 96.15% (25/26) were infected with HPV and 84.62% (22/26) were HR-HPV. The overall positivity of each and co-testing between cytology or HPV DNA testing or p16/Ki-67 dual staining was evaluated. In each cervical lesion, primary HPV DNA testing showed the highest sensitivity, but low specificity. The combined all HPV/HR-HPV with p16/Ki-67 detection increased the specificity of abnormal cervical lesions. Conclusion: P16/Ki-67 dual stain cytology in HPV-positive women performs well for diagnosis of abnormal cervical lesions and should be considered for management of HPV-positive women to avoid unnecessary colposcopy referrals.     

Human papillomavirus testing using hybrid capture II with SurePath collection: initial evaluation and longitudinal data provide clinical validation for this method

BACKGROUND: Testing for human papillomavirus (HPV) is an integral part of equivocal cervical cytology triage. Clinical validation of non-FDA (Food and Drug Administration)-approved methods is therefore important because of the high volume of such tests and the implications for missed high-grade lesions if test performance is not optimal. METHODS: A preinitiation study and 17 months of follow-up data using Hybrid Capture II (HC II) HPV detection with SurePath (SP) sample collection were analyzed. Results of HPV tests on abnormal cytology samples were collected and compared with follow-up results. HPV-positive rates were determined in cases of low-grade squamous intraepithelial lesion (LSIL) and high-grade squamous intraepithelial lesion (HSIL), and follow-up rates of cervical intraepithelial neoplasia (CIN) were determined in HPV-positive and -negative cases of atypical squamous cells of unknown significance (ASC-US). Rates were compared with published data using FDA-validated methods. RESULTS: The preinitiation study showed the test method to be 100% sensitive for the detection of LSIL (20 cases) and HSIL (8). The ASC-US follow-up study (2319 cases with 625 having biopsy results) showed that the rate of CIN III+ in HPV +/- cases was 7.8%/1.4%, and of CIN II+ was 17.5%/4.3%, respectively. The positive predictive values/negative predictive values (PPV/NPVs) (CIN II+) for the test were 17.5%/95.7%, respectively. CONCLUSIONS: Published FDA-validated HPV testing follow-up data show that the expected rates of CIN III+ and CIN II+ in the HPV-negative ASC-US population are 1.4% and 5%, respectively, with PPV/NPVs (CIN II+) of 20%/99%, respectively. By comparison, the present data using HC II with SP show strong similarity, indicating clinical validity for the use of this method.     

Combined use of cytology,p16 immunostaining and genotyping for triage of women positive for high-risk human papillomavirus at primary screening

Human papillomavirus (HPV) testing is very sensitive for primary cervical screening but has low specificity. Triage tests that improve specificity but maintain high sensitivity are needed. Women enrolled in the experimental arm of Phase 2 of the New Technologies for Cervical Cancer randomized controlled cervical screening trial were tested for high-risk HPV (hrHPV) and referred to colposcopy if positive. hrHPV-positive women also had HPV genotyping (by polymerase chain reaction with GP5+/GP6+ primers and reverse line blotting), immunostaining for p16 overexpression and cytology. We computed sensitivity, specificity and positive predictive value (PPV) for different combinations of tests and determined potential hierarchical ordering of triage tests. A number of 1,091 HPV-positive women had valid tests for cytology, p16 and genotyping. Ninety-two of them had cervical intraepithelial neoplasia grade 2+ (CIN2+) histology and 40 of them had CIN grade 3+ (CIN3+) histology. The PPV for CIN2+ was >10% in hrHPV-positive women with positive high-grade squamous intraepithelial lesion (61.3%), positive low-grade squamous intraepithelial lesion (LSIL+) (18.3%) and positive atypical squamous cells of undetermined significance (14.8%) cytology, p16 positive (16.7%) and, hierarchically, for infections by HPV33, 16, 35, 59, 31 and 52 (in decreasing order). Referral of women positive for either p16 or LSIL+ cytology had 97.8% sensitivity for CIN2+ and women negative for both of these had a 3-year CIN3+ risk of 0.2%. Similar results were seen for women being either p16 or HPV16/33 positive. hrHPV-positive women who were negative for p16 and cytology (LSIL threshold) had a very low CIN3+ rate in the following 3 years. Recalling them after that interval and referring those positive for either test to immediate colposcopy seem to be an efficient triage strategy. The same applies to p16 and HPV16.     

Effectiveness of two strategies to follow-up ASC-US and LSIL screening results in The Netherlands using repeat cytology with or without additional hrHPV testing: a retrospective cohort study

Purpose

The purpose of the study was to assess the effectiveness of repeat cytology with and without additional high-risk human papilloma virus (hrHPV) testing after atypical squamous cells of undetermined significance/low-grade squamous intraepithelial lesion (ASC-US/LSIL) screening results.

Methods

In the Netherlands, ASC-US/LSIL is triaged by repeat cytology at 6 months or repeat cytology at 6 months with additional hrHPV testing. ASC-US/LSIL results from 13,734 screenees in 2008 were extracted from “Dutch Pathology Registry” including cytology, histology, and/or HPV follow-up results. Proportions of compliance, repeat cytology, referral, and detected cervical intraepithelial neoplasia (CIN) were assessed.

Results

With additional hrHPV testing, 46.8 % was send back to regular screening at 6 months, 28.6 % needed second repeat cytology, and 24.6 % was referred for colposcopy. Without additional hrHPV testing, this was 0.0, 76.1, and 23.9 %, respectively. With additional hrHPV testing, significantly higher proportions of persisting ASC-US/LSIL; compliance with repeat/referral advices; and histological detection of CIN0 (no CIN or cancer), CIN1, and CIN2 were found but equal proportions CIN3+.

Conclusions

Additional hrHPV testing shortens follow-up without altering CIN3+ detection. Detection of CIN0, CIN1, and CIN2 was higher, presumably by hrHPV-driven biased cytology and detection bias. Restricting additional hrHPV testing to older women, reading cytology without knowledge of hrHPV status, and addition of more specific triage tests could further improve the effectiveness of additional hrHPV testing.     

Performance of HPV DNA Testing with Hybrid Capture 2 in Triaging Women with Minor Cervical Cytologic Abnormalities (ASC-US/LSIL) in Northern Thailand

Background: Minor cervical cytologic abnormalities include atypical squamous cells of undeterminedsignificance (ASC-US) and low-grade squamous intraepithelial lesion (LSIL). Approximately 10-20% of womenwith minor cytologic abnormalities have histologic high-grade squamous intraepithelial or worse lesions (HSIL+).In Thailand, women with minor cytologic abnormalities have a relatively high risk of cervical cancer, and referralfor colposcopy has been suggested. A triage test is useful in the selection of women at risk for histologic HSIL+ toreduce the colposcopy burden. The aim of this study was to assess the performance of high-risk HPV DNA test intriage of women with minor cytologic abnormalities in northern Thailand. Materials and Methods: All womenwith ASC-US/LSIL cytology who were referred to our colposcopy clinic from October 2010 to February 2014were included. HPV DNA testing was performed using Hybrid Capture 2 (HC2). All patients received colposcopicexamination. Accuracy values of HC2 in predicting the presence of histologic HSIL+ were calculated. Results:There were 238 women in this study (121 ASC-US and 117 LSIL). The HC2 positivity rate was significantlyhigher in the LSIL group than in ASC-US group (74.8% versus 41.0%, p<0.001). Histologic HSIL+ was detectedin 9 women (7.4%) in the ASC-US group and 16 women (13.7%) in the LSIL group (p=0.141). There was nohistologic HSIL+ detected among HC2-negative cases (sensitivity and negative predictive value of 100%). Theperformance of HC2 triage was highest among women aged >50 years with ASC-US cytology. An increase in thecut-off threshold for positive HC2 resulted in a substantial decrease of sensitivity and negative predictive value.Conclusions: HPV DNA testing with HC2 shows very high sensitivity and negative predictive value in triage ofwomen with minor cervical cytologic abnormalities in northern Thailand. An increase of the cut-off thresholdfor HC2 triage is not recommended in this region.     

A cohort study of cervical screening using partial HPV typing and cytology triage

HPV testing is more sensitive than cytology for cervical screening. However, to incorporate HPV tests into screening, risk‐stratification (“triage”) of HPV‐positive women is needed to avoid excessive colposcopy and overtreatment. We prospectively evaluated combinations of partial HPV typing (Onclarity, BD) and cytology triage, and explored whether management could be simplified, based on grouping combinations yielding similar 3‐year or 18‐month CIN3+ risks. We typed ～9,000 archived specimens, taken at enrollment (2007–2011) into the NCI‐Kaiser Permanente Northern California (KPNC) HPV Persistence and Progression (PaP) cohort. Stratified sampling, with reweighting in the statistical analysis, permitted risk estimation of HPV/cytology combinations for the 700,000+‐woman KPNC screening population. Based on 3‐year CIN3+ risks, Onclarity results could be combined into five groups (HPV16, else HPV18/45, else HPV31/33/58/52, else HPV51/35/39/68/56/66/68, else HPV negative); cytology results fell into three risk groups (“high‐grade,” ASC‐US/LSIL, NILM). For the resultant 15 HPV group‐cytology combinations, 3‐year CIN3+ risks ranged 1,000‐fold from 60.6% to 0.06%. To guide management, we compared the risks to established “benchmark” risk/management thresholds in this same population (e.g., LSIL predicted 3‐year CIN3+ risk of 5.8% in the screening population, providing the benchmark for colposcopic referral). By benchmarking to 3‐year risk thresholds (supplemented by 18‐month estimates), the widely varying risk strata could be condensed into four action bands (very high risk of CIN3+ mandating consideration of cone biopsy if colposcopy did not find precancer; moderate risk justifying colposcopy; low risk managed by intensified follow‐up to permit HPV “clearance”; and very low risk permitting routine screening.) Overall, the results support primary HPV testing, with management of HPV‐positive women using partial HPV typing and cytology.     

Risk-stratification of HPV-positive women with low-grade cytology by FAM19A4/miR124-2 methylation and HPV genotyping

Background The introduction of primary HPV screening has doubled the number of colposcopy referrals because of the direct referral of HPV-positive women with a borderline or mild dyskaryosis (BMD) cytology (ASC-US/LSIL) triage test. Further risk-stratification is warranted to improve the efficiency of HPV-based screening.Methods This study evaluated the discriminative power of FAM19A4/miR124-2 methylation, HPV16/18 genotyping and HPV16/18/31/33/45 genotyping in HPV-positive women with BMD (n = 294) in two Dutch screening trials. Absolute CIN3+ risks and colposcopy referrals within one screening round were calculated.Results Methylation analysis discriminated well, yielding a CIN3+ risk of 33.1% after a positive result and a CIN3+ risk of 9.8% after a negative result. HPV16/18 and HPV16/18/31/33/45 genotyping resulted in a 27.6% and 24.6% CIN3+ risk after a positive result, and a 13.2% and 9.1% CIN3+ risk after a negative result. Colposcopy referral percentages were 41.2%, 43.2%, and 66.3% for FAM19A4/miR124-2 methylation, HPV16/18 and HPV16/18/31/33/45 genotyping, respectively. The CIN3+ risk after a negative result could be lowered to 2.8% by combining methylation and extended genotyping, at the expense of a higher referral percentage of 75.5%.Conclusion The use of FAM19A4/miR124-2 methylation and/or HPV genotyping in HPV-positive women with BMD can lead to a substantial reduction in the number of direct colposcopy referrals.Subject terms: DNA methylation, Diagnostic markers, Cervical cancer, Molecular medicine     

Atypical squamous cells, cannot exclude high-grade squamous intraepithelial lesion: diagnostic performance, human papillomavirus testing, and follow-up results

BACKGROUND: Current guidelines recommend colposcopy rather than high-risk human papillomavirus (HPV) testing for the evaluation of abnormal cervical cytology interpreted as "atypical squamous cells, cannot exclude high-grade squamous intraepithelial lesion" (ASC-H) based on data from the Atypical Squamous Cells of Undetermined Significance/Low-Grade Squamous Intraepithelial Lesion (ASCUS/LSIL) Triage Study (ALTS), which indicated that ASC-H had a significantly greater frequency of high-risk HPV positivity and underlying high-grade squamous intraepithelial lesions (HSIL) compared with ASCUS. The cytologic interpretations in the ALTS were expert consensus diagnoses rather than routine, single-pathologist readings. METHODS: The authors conducted a comparative analysis of Hybrid Capture 2 high-risk HPV positivity and frequency of histologically diagnosed HSIL for ASC-H and ASCUS to evaluate the performance of ASC-H as a cytologic interpretation subcategory and the potential utility of HPV testing for colposcopy triage of ASC-H in routine practice. RESULTS: Sixty-four of 96 patients with ASC-H (66.7%) were HPV-positive compared with 484 of 1079 patients with ASCUS (44.9%). Among the patients who had histologic follow-up, HSIL was identified in 18 of 45 patients (40.0%) with HPV-positive ASC-H compared with 27 of 266 patients (10.2%) with HPV-positive ASCUS (P < 0.0001) and 1 of 22 patients (4.5%) with HPV-negative ASC-H (P = 0.003); the latter result was similar to the finding of HSIL in 5 of 85 patients (5.9%) with HPV-negative ASCUS. The frequency of HPV-positive ASC-H in the current study (67%) was lower than that obtained in the ALTS for ASC-H (86%) but higher than that for ASCUS in both this study (45%) and in the ALTS (51% for all ASC; 63% for ASCUS, equivocal for LSIL). Underlying HSIL was detected in a similar percentage of patients with HPV-positive ASC-H in this study and in the ALTS (41%). CONCLUSIONS: The greater frequency of HPV positivity and the significantly increased risk of underlying HSIL for ASC-H compared with ASCUS indicated that ASC-H category utilization and performance are appropriate in this routine clinical practice setting. The lower frequency of HPV positivity for ASC-H compared with the ALTS data and the similar low risk of HSIL in HPV-negative ASC-H and HPV-negative ASCUS indicate that HPV testing for triage of ASC-H in routine practice has the potential to reduce the number of women who are referred for colposcopy without an increased risk of failure to detect HSIL among HPV-negative women, similar to its triage role for ASCUS.     

Eight-type human papillomavirus E6/E7 oncoprotein detection as a novel and promising triage strategy for managing HPV-positive women

The management of HPV-positive women becomes particularly crucial in cervical cancer screening. Here we assessed whether detection of E6 or E7 oncoproteins targeting eight most prevalent HPV types could serve as a promising triage option. Women (N = 1,416) aged 50–60 from Shanxi, China underwent screening with HPV testing and liquid-based cytology (LBC), with any positive results referring to colposcopy and biopsy if necessary. Women with HPV-positive results received further tests using DNA-based genotyping, E6 or E7 oncoprotein detection targeting HPV16/18 (for short: E6 (16/18) Test) or HPV16/18/31/33/35/45/52/58 (for short: E6/E7 (8 types) Test), respectively. Among HPV-positive women, E6/E7 (8 types) oncoproteins had lower positivity (17.37%) compared to DNA-based genotyping for same eight types (58.30%) and LBC with ASC-US threshold (50.97%); HPV16 was the genotype showing the highest frequency (8.49%) for oncoprotein detection followed by HPV52 (3.47%), 58 (2.32%), 33 (1.54%), 18 (1.16%), 45 (0.77%), 35 (0.39%) and 31 (0%). For detection of cervical intraepithelial neoplasia Grade 3 or higher (CIN3+), E6/E7 (8 types) Test had similar sensitivity (100.00%) and superior specificity (85.94%) as well as positive predictive value (PPV, 22.22%) compared to both LBC and DNA-based genotyping (8 types); For detection of CIN2+, E6/E7 (8 types) Test was less sensitive (67.74%) but still more specific (89.47%) and risk predictive with PPV of 46.67%. Notably, E6/E7 (8 types) Test remarkably decreased the number of colposcopies needed to detect one CIN2+ and CIN3+ (2.14 and 4.50). E6/E7 oncoprotein detection showed a good “trade-off” between sensitivity and specificity with more efficient colposcopy referrals, which is of great importance to maximize the benefits of HPV-based screening program, especially applicable for the areas with high HPV prevalence and low-resources.     

Management of HPV-positive women in cervical screening using results from two consecutive screening rounds

We studied whether triage of human papillomavirus (HPV)-positive women participating in an HPV-based screening programme can be improved by including the HPV result at the previous screen in the triage algorithm. We analyzed data of a subgroup of 366 women from the POBASCAM trial, screened by cytology and HPV cotesting. Women were included if they tested HPV-positive in the second HPV-based screening round. We evaluated the clinical performance of 16 strategies, consisting of cytology, HPV genotyping, and/or previous screen HPV result. The clinical endpoint was cervical precancer or cancer (CIN3+). The current Dutch triage testing policy for HPV-positive women is to refer women for colposcopy if they have abnormal cytology at baseline or after 6–18 months. In the second HPV-based screening round, this strategy yielded a negative predictive value (NPV) of 95.8% (95% confidence interval: 91.9–98.2) and colposcopy referral rate of 37.6% (32.3–43.2%). Replacing repeat cytology by the previous screen HPV result yielded a similar NPV (96.9%, 93.3–98.9) and colposcopy referral rate (38.8%, 33.4–44.4). A higher NPV (99.2%, 96.3–100%) at the cost of a higher colposcopy referral rate (49.2%, 43.6–54.8) was achieved when cytology was combined with HPV16/18 genotyping. The other 13 triage strategies yielded a lower NPV, a higher colposcopy referral rate or performed similarly but required additional testing. HPV-positive women in the second HPV-based screening round can be suitably managed by cytology, HPV16/18 genotyping and the HPV result at the previous screen, obviating the need for repeat testing of HPV-positive, cytology negative women.     

Assessment of reflex human papillomavirus DNA testing in patients with atypical endocervical cells on cervical cytology

BACKGROUND: Reflex human papillomavirus (HPV) testing for atypical squamous cells of undetermined significance (ASC-US) has improved the sensitivity and specificity in detecting high-grade squamous dysplasia (cervical intraepithelial neoplasia [CIN]2+). However, to the authors' knowledge there are no guidelines for performing reflex HPV testing in women with atypical endocervical cells (AEC) before colposcopy. This report is of a 5-year experience with reflex HPV testing in women with AEC and assessment of the potential role of reflex HPV testing in guiding subsequent colposcopy-directed cervical biopsy/curettage in a large tertiary care hospital setting. METHODS: All AEC cases cytologically diagnosed from July 2001 to June 2006 were retrieved from the Cleveland Clinic database. The histopathologic diagnoses and the results of HPV testing using the Hybrid Capture 2 (HC-II) method were reviewed. The most severe histopathologic diagnosis was recorded. RESULTS: Of a total 332,470 Papanicolaou (Pap) tests performed, 317 cases of AEC had histopathologic follow-up and reflex testing for high-risk HPV. Histopathologic examination of the 64 HPV-positive AEC cases revealed 18 cases of endocervical adenocarcinoma in situ/adenocarcinoma (AIS+) and 22 cases of CIN2+. Among 253 of the HPV-negative AEC women, AIS+ was found in only 3 cases and CIN2+ in 1 case. Cervical AIS+ was found in 28% of the HPV-positive AEC patients and in only 0.9% of the HPV-negative patients (P<.0001). When the significant glandular (AIS+) and squamous (CIN2+) lesions were combined, 62.5% of the lesions were detected in HPV-positive AEC cases compared with 1.6% in the HPV-negative AEC cases (P<.0001). CONCLUSIONS: Because of a high sensitivity (91.0%) and high specificity (91.2%) in detecting significant cervical lesions, reflex HPV testing for cytologic diagnosis of AEC appears to be a useful ancillary tool in the selection of high-risk patients for colposcopy.     

宫颈癌多种筛查方案的研究

目的 探索适宜我国不同地区的宫颈癌筛查方案,以提高我国妇女宫颈癌的防治水平.方法 利用1999年在山西省襄垣县开展的一项以人群为基础的宫颈癌筛查横断面研究的资料,所有筛查对象均进行了薄层液基细胞学(LBC)、荧光镜检、醋酸染色法(VIA)、阴道镜检查、自我取样人乳头瘤病毒(HPv)检测和医生取样HPV检测等6种宫颈癌筛查方法 ,而且每位筛查对象均有病理诊断结果 .采用筛查试验的串、并联法组合各种筛查技术,比较所得方案识别宫颈高度以上病变[≥宫颈上皮内瘤变(CIN)2]的灵敏度、特异度和阴道镜转诊率等指标,以受试者工作特征曲线(ROC)下面积综合分析各筛查方案.结果 LBC检测以未明确意义的不典型鳞状细胞(ASC-US)为阳性,HPV检测以HPV DNA≥1.0 ps/mi为阳性.在LBC和HPV检测组合方案中,并联初筛方法 (即两者任一项阳性即判断为筛查阳性)的灵敏度为100.O%,特异度为68.6%,阴道镜转诊率为34.4%;LBC初筛HPV分流方法 (即ASC-US者进行HPV检测)的灵敏度为93.0%,特异度为89.9%,阴道镜转诊率为13.7%;HPV初筛LBC分流方法 (即 HPV阳性者进行LBC检测)的灵敏度为91.7%,特异度为93.0%,阴道镜转诊率为10.6%.经ROC分析,LBC初筛HPV分流方法 和HPV初筛LBC分流方法 明显优于单纯并联初筛方法 (P=0.0003;P=0.0002).单独以LBC或HPV检测作为筛查方案时,以ASC-US或低度病变(LSIL)为筛查阳性的LBC方法 灵敏度、特异度和阴道镜转诊率分别为94.2%、77.3%、25.7%和87.2%、93.5%、10.O%;医生取样HPV检测方法 和自我取样HPV检测方法 的灵敏度、特异度和阴道镜转诊率分别为97.6%、84.8%、18.8%和83.5%、85.9%、17.1%.经ROC分析,医生取样HPV检测方法 优于以ASC-US为筛查阳性的LBC方法 或自我取样HPV检测方法 (P=0.005,P=0.002).在VIA及其与HPV检测的组合方案中,单独采用VIA筛查方法 的灵敏度为70.9%,特异度为74.3%,阴道镜转诊率为27.6%;HPV初筛VIA分流方法 (即自我取样HPV检测阳性者进行VIA检查)的灵敏度、特异度和阴道镜转诊率分别为65.9%、95.2%和7.4%.经ROC分析,HPV初筛VIA分流方法 明显优于单独使用VIA方法 (P=0.004).结论 根据地区资源条件和个人意愿,我国经济发达地区可选用HPV初筛LBC分流方法 或LBC初筛HPV分流方法 作筛查手段;中等经济发展水平的中小城市可选用单独以LBC或HPV检测方法 作为筛查手段;VLA是欠发达地区可行的筛查方法 ,在廉价HPV检测试剂盒上市后,可选择HPV初筛VIA分流方法 ,以进一步提高宫颈癌的筛查效力.     

Cost-Effectiveness Analysis of Different Management Strategies for Detection CIN2+ of Women with Atypical Squamous Cells of Undetermined Significance (ASC-US) Pap Smear in Thailand

Background: To identify the optimal cost effective strategy for the management of women having ASC-USwho attended at King Chulalongkorn Memorial Hospital (KMCH). Design: An Economical Analysis based ona retrospective study. Subject: The women who were referred to the gynecological department due to screeningresult of ASC-US at King Chulalongkorn Memorial Hospital, a general and tertiary referral center in BangkokThailand, from Jan 2008 - Dec 2012. Materials and Methods: A decision tree-based was constructed to evaluatethe cost effectiveness of three follow up strategies in the management of ASC-US results: repeat cytology, triagewith HPV testing and immediate colposcopy. Each ASC-US woman made the decision of each strategy afterreceiving all details about this algorithm, advantages and disadvantages of each strategy from a doctor. The modelcompared the incremental costs per case of high-grade cervical intraepithelial neoplasia (CIN2+) detected asmeasured by incremental cost-effectiveness ratio (ICER). Results: From the provider’s perspective, immediatecolposcopy is the least costly strategy and also the most effective option among the three follow up strategies.Compared with HPV triage, repeat cytology triage is less costly than HPV triage, whereas the latter providesa more effective option at an incremental cost-effectiveness ratio (ICER) of 56,048 Baht per additional case ofCIN 2+ detected. From the patient’s perspective, the least costly and least effective is repeat cytology triage.Repeat colposcopy has an incremental cost-effectiveness (ICER) of 2,500 Baht per additional case of CIN2+detected when compared to colposcopy. From the sensitivity analysis, immediate colposcopy triage is no longercost effective when the cost exceeds 2,250 Baht or the cost of cytology is less than 50 Baht (1USD = 31.58 THB).Conclusions: In women with ASC-US cytology, colposcopy is more cost-effective than repeat cytology or triagewith HPV testing for both provider and patient perspectives.     

Impact of HPV testing in opportunistic cervical screening: Support for primary HPV screening in the United States

Human papillomavirus (HPV) testing for cervical screening increases diagnosis of precancer and reduces the incidence of cervical cancer more than cytology alone. However, real-world evidence from diverse practice settings is lacking for the United States (U.S.) to support clinician adoption of primary HPV screening. Using a population-based registry, which captures all cervical cytology (with or without HPV testing) and all cervical biopsies, we conducted a real-world evidence study of screening in women aged 30 to 64 years across the entire state of New Mexico. Negative cytology was used to distinguish cotests from reflex HPV tests. A total of 264 198 cervical screening tests (with exclusions based on clinical history) were recorded as the first screening test between 2014 and 2017. Diagnoses of cervical intraepithelial neoplasia grades 2 or 3 or greater (CIN2+, CIN3+) from 2014 to 2019 were the main outcomes. Of cytology-negative screens, 165 595 (67.1%) were cotests and 4.8% of these led to biopsy within 2 years vs 3.2% in the cytology-only group. Among cytology-negative, HPV tested women, 347 of 398 (87.2%) CIN2+ cases were diagnosed in HPV-positive women, as were 147 of 164 (89.6%) CIN3+ cases. Only 29/921 (3.2%) CIN3+ and 67/1964 (3.4%) CIN2+ cases were diagnosed in HPV-negative, cytology-positive women with biopsies. Under U.S. opportunistic screening, across a diversity of health care delivery practices, and in a population suffering multiple disparities, we show adding HPV testing to cytology substantially increased the yield of CIN2+ and CIN3+. CIN3+ was rarely diagnosed in HPV-negative women with abnormal cytology, supporting U.S. primary HPV-only screening.     

高危型HPV E6/E7mRNA检测在宫颈机会性筛查中应用价值研究

目的 人乳头瘤病毒(human papillomavirus,HPV) DNA能提高检测宫颈上皮内瘤变2级+(cervical intraepithelial neoplasia 2+,CIN2+)的灵敏度,但也增加了一过性HPV感染的检出率.本研究应用高危型HPV(high risk human papillomavirus,hrHPV) E6/E7 mRNA对比二代杂交捕获(hybrid CaptureⅡ,HC2)、HPV分型,描述机会性筛查人群中HPV感染及宫颈病变分布特征,探讨其用于宫颈筛查的可行性.方法 选取2013-01-01-2015-12-31在青岛大学附属青岛市立医院(6 138例)和青岛市城阳区人民医院(1 653例)妇科门诊行宫颈液基细胞学筛查的女性共计7 791例,年龄21～65岁.所有筛查女性行液基细胞学检查的同时行HPV检测,根据HPV检测方法的不同分为HC2组、HPV分型组和E6/E7组共3组.计算各组细胞学结果异常率、HPV阳性率和宫颈病变检出率,比较E6/E7所描述的流行病学特征与HPV-DNA方法的不同.结果 E6/E7组各级细胞学结果分别为,未见上皮内瘤变及恶性病变(negative for intraepithelial lesion or malignancy,NILM) 87.51％,意义不明的非典型鳞状细胞(atypical squamous cells of undetermined significance,ASCUS)5.47％,低级别鳞状上皮内瘤变(low-grade squamous intraepithelial lesion,LSIL)3.51％,不除外高级别鳞状上皮内瘤变的非典型细胞(atypical squamous cells cannot exclude HSIL,ASC-H)0.52％,高级别鳞状上皮内瘤变(high-grade squamous intraepithelial lesion,HSIL)2.99％.HPV阳性中异常细胞学结果的比例明显高于HPV阴性,而NILM的比例低于HPV阴性,P＜0.001.随着细胞学异常程度的加重,HPV阳性率逐渐增高,差异有统计学意义,x2值分别为611.089、512.036和767.260,P值均＜0.001.E6/E7组中NILM的HPV阳性率为8.02％,合计阳性率为14.73％,均较HC2及分型组低,x2=30.174,P=0.000;x2 =22.991,P＜0.001. E6/E7组CIN2+/CIN3+检出率分别为5.92％和1.20％,3组间比较差异无统计学意义,x2值分别为1.499和0.711,P值分别为0.473和0.701.E6/E7阳性率在正常至浸润性宫颈癌(invasive cervical cancer,ICC)之间随病变程度加重而升高,P＜0.05.在正常病理结果中E6/E7阳性率为61.36％,低于HC2和HPV分型,x2=15.767,P＜0.001;其余病理结果中各组间HPV阳性率差异无统计学意义,P＞0.05.结论 hrHPV E6/E7 mRNA较之HC2和HPV分型能在不降低宫颈病变检出率的前提下减少一过性HPV感染的检出率,提示其在宫颈筛查中具有潜在应用价值.     

Comparison of three management strategies for patients with atypical squamous cells of undetermined significance: baseline results from a randomized trial

BACKGROUND: More than 2 million U.S. women receive an equivocal cervical cytologic diagnosis (atypical squamous cells of undetermined significance [ASCUS]) each year. Effective colposcopy triage strategies are needed to identify the minority of women who have clinically significant disease while avoiding excessive follow-up evaluation for others. METHODS: The ASCUS/LSIL (i.e., low-grade squamous intraepithelial lesion) Triage Study (ALTS) is a multicenter, randomized trial comparing the sensitivity and specificity of the following three management strategies to detect cervical intraepithelial neoplasia grade 3 (CIN3): 1) immediate colposcopy (considered to be the reference standard), 2) triage to colposcopy based on human papillomavirus (HPV) results from Hybrid Capture 2(TM) (HC 2) and thin-layer cytology results, or 3) triage based on cytology results alone. This article summarizes the cross-sectional enrollment results for 3488 women with a referral diagnosis of ASCUS. All statistical tests are two-sided. RESULTS: Among participants with ASCUS, the underlying prevalence of histologically confirmed CIN3 was 5.1%. Sensitivity to detect CIN3 or above by testing for cancer-associated HPV DNA was 96.3% (95% confidence interval [CI] = 91.6% to 98.8%), with 56.1% of women referred to colposcopy. Sensitivity of a single repeat cytology specimen with a triage threshold of HSIL or above was 44.1% (95% CI = 35.6% to 52.9%), with 6.9% referred. Sensitivity of a lower cytology triage threshold of ASCUS or above was 85.3% (95% CI = 78.2% to 90.8%), with 58.6% referred. CONCLUSIONS: HC 2 testing for cancer-associated HPV DNA is a viable option in the management of women with ASCUS. It has greater sensitivity to detect CIN3 or above and specificity comparable to a single additional cytologic test indicating ASCUS or above.     

Test positivity cutoff level of a high risk human papillomavirus test could be increased in routine cervical cancer screening

We present data on test positivity, relative sensitivity, rates of detection and relative specificity for primary human papillomavirus (HPV) testing with different cutoff levels for test positivity, in comparison to conventional cytology. In 2003-2004, 18,438 women were screened primarily with Hybrid Capture 2 (HC 2) assay, a test for oncogenic HPV DNA, and 21,446 with conventional cytology within the organised screening programme in Finland. A cytological triage test was performed for the HPV positives. Women with cytology equal to low grade squamous intraepithelial lesion (LSIL) or worse were referred for colposcopy. The relative sensitivity measured as relative risk (RR) of any cervical intraepithelial neoplasia (CIN) or cancer was 1.58 for the HPV test at the relative light units (rlu) ratio cutoff 1.00, in comparison to cytology. With the cutoff 3.00, all CIN 2+ lesions were detected. With cutoff 10.00, 2 of the 22 CIN 3+ lesions were missed. Relative specificity for HPV screening for any CIN was 92.6% at cutoff 1.00, 94.6% at cutoff 3.00 and 96.3% at cutoff 10.00. For CIN 3+ specificity estimates for these cutoffs were 92.1%, 94.1% and 95.8%, respectively. Used for routine screening as the sole test, the HPV test cutoff can be increased from the level recommended for clinical use. With HC 2, the detection rate at rlu ratio cutoff 10.00 is still at the level of high-quality conventional screening. At that level, the false positive rate is reduced by about half and the specificity of the HPV test becomes equal to the average specificity of conventional cytology.     

Human papillomavirus testing for triage of women with low‐grade squamous intraepithelial lesions

Low‐grade squamous intraepithelial lesion (LSIL) is a common cytologic finding in cervical screening, yet only about 10–20% have significant histologic abnormalities and these are almost always positive for high‐risk human papillomavirus (hrHPV). This analysis aims to clarify the role of hrHPV DNA testing in the triage of women with LSIL cytology. In the ATHENA screening trial, we examined 1,084 cases of LSIL, of which 925 had an evaluable biopsy, to determine the extent to which hrHPV testing can identify those patients who have precursor lesions in need of immediate clinical referral and those who have changes more likely to regress spontaneously. Overall, 71.2% of LSIL cases were hrHPV positive, but the prevalence was age dependent, with only 56.1% in women ≥40 years. Among women with LSIL, 11.6% (107/925) had a cervical intraepithelial neoplasia grade 2 or worse (CIN2+) histologic diagnosis and, of these, only nine were hrHPV negative. For CIN3+, 91.7% (44/48) of women with LSIL were hrHPV positive. The negative predictive value of hrHPV testing for CIN3+ in LSIL was 100% for women aged ≥40 years. Women who were HPV16 positive had a higher positive predictive value for CIN2+ (25.4%) than those who were positive for 12 other pooled hrHPV types (11.5%). Testing for hrHPV in women with LSIL is effective in identifying high‐grade cervical lesions, thereby avoiding unnecessary referrals to colposcopy and potential over‐treatment of non‐progressive lesions, especially for women aged ≥40 years.     

Silent High Grade Cervical Intraepithelial Neoplasia in Atypical Smears from Liquid Based Cervical Cytology - Three Years Experience in Thammasat University Hospital

Purpose: To study the prevalence of CIN2 diagnosis in women with atypical Papaniculoau (Pap) smears to suggest appropriate management option for Thai health care. Materials and Methods: Data from all patients with liquid based cytology with human papillomavirus (HPV) testing between May 2013 - May 2016 were collected from medical records. Women with atypical cervical Pap smears were recruited. Results for age, HPV testing, HPV 16, 18, 45 and other genotypes tested, colposcopic examination and histopathological assessment were all collected. Atypical smears were defined as atypical squamous cells of undetermined significance (ASC-US) and atypical squamous cells cannot be exclude high grade squamous intraepithelial lesion (ASC-H). Results: A total of 2,144 cases were recruited. Twenty six women with ASC-US on cytology had high risk (HR) HPV detection while eight cases with ASC-H had HR-HPV (40.0% VS 72.7%, p0.005). Among the 26 women with ASC-US cytology and positive HR-HPV, HPV type 16 (n8, 30.8%), type 18 (n1, 3.8%), type 45 (n1, 3.8%) and other HPV types (n17, 65.4%) were found. Eight women with ASC-H and positive HR-HPV demonstrated type 16 (n6, 75%) and other HPV types (n2, 25%). Fifty seven women with ASC-US had normal colposcopy, CIN1 and CIN2 at percentages of 80.7 (46/57), 14.0 (8/57) and 5.3 (3/57), respectively. In the ASC-H group, 7 out of 10 women had normal colposcopy and three (30%) had CIN2 results. Conclusions: In women with ASC-US cytology, immediate colposcopy is highly recommended. HPV testing can be performed if colposcopy is not an available option because there was high prevalence (5.3%) of CIN2 in our findings. ASCCP recommendations for ASC-H that colposcopy should be performed on all ASC-H cases regardless of HPV result are thereby supported by the findings of this investigation.     

Cost and Effectiveness Comparison of Immediate Colposcopy Versus Human Papillomavirus DNA Testing in Management of Atypical Squamous Cells of Undetermined Significance in Turkish Women

Background: A small but significant proportion of cases with atypical squamous cells of undeterminedsignificance (ASCUS) may harbour CIN 2-3, or even invasive carcinoma. Although immediate colposcopy,HPV-DNA testing or expectant management are three recommended options in ASCUS triage, a consensus doesnot currently exist on which one of these approaches is the most efficient. In this study, we aimed to comparethe performance and cost of immediate colposcopy and colposcopy based on the human papillomavirus (HPV)testing for detecting histologically confirmed high-grade cervical intraepithelial neoplasia (CIN) in women withASCUS. Materials and Methods: Records of 594 women with an index Papanicolaou smear showing ASCUSwere retrospectively analyzed. Women in the immediate colposcopy arm were referred directly to colposcopy(immediate colposcopy group, n=255) and those in the HPV triage arm were proceeded to colposcopy if thehigh-risk HPV (hrHPV) test was positive (HPV triage group, n=339). High grade CIN (CIN2+) detection rateand treatment costs were compared between the groups. Results: The detected rate of CIN2+ was higher inthe HPV triage group compared to immediate colposcopy group (8% vs. 1.6%, p=0.011). In the HPV triagegroup, the total cost, cost per patient, and the cost for detecting one case of high grade CIN were higher thanthe immediate colposcopy group (p<0.001). Conclusions: In women with ASCUS cytology, HPV DNA testingfollowed by colposcopy is more costly than immediate colposcopy, but this approach is associated with a higherrate of CIN2+ detection. This findings suggest that HPV DNA testing combined with cervical cytology couldreduce the referral rate to colposcopy