- Email: [email protected]
Maximizing the Adequacy of Cervista® HPV Results on ThinPrep® Pap Samples Treated with Glacial Acetic Acid
S38
Abstracts
Graph of Table 2.
66 Molecular Testing and Cervical Screening: Will One Test Fit All? Erin McCarthy, BS, CT(ASCP)1, Changhong Ye, BS, SCT(ASCP)2, Daniel Kurtycz, MD2. 1University of Wisconsin, Madison, Wisconsin; 2 Wisconsin State Laboratory of Hygiene, Madison, Wisconsin
screening tool, we compared the risk levels of high-grade cervical intraepithelial neoplasia (CIN2+) during a 3-year follow-up period in women aged 30 years and older who screened with HPV and Pap co-testing. Materials and Methods: Patient records from the Cancer Prevention Center were searched retrospectively for women aged 30 years and older who underwent cervical cancer screening during 2007 and 2008. Screening consisted of SurePath Pap and Hybrid Capture 2 (HC2) HPV co-testing. Cumulative incidences of CIN2+ were compared to evaluate the risks stratified by HPV and/or Pap testing results. Results: During the study period, 5,739 women underwent screening. HPV was positive in 3.75% (215/5,739) of the women. Of the 2,107 women for whom 3-year follow-up data were available, 189 (9.0%) had a positive HPV result and 188 (8.9%) had a positive Pap result (ASC-US+). During the 3year follow-up period, 20 women had a CIN2+ biopsy result (10 CIN2 cases, 10 CIN3 cases). The cumulative risk of CIN2+ in women with a positive HPV result (8.5%) was significantly higher than that in women with a negative HPV result (0.26%; P Conclusions: During a 3-year screening interval, the risk of CIN2+ in women aged 30 years and older with a HPV-negative result is sufficiently low to ensure a safety margin for cervical cancer prevention, supporting the use of HPV testing as a first-line screening tool for cervical cancer prevention in women aged 30 and older.
Introduction: Over time, cervical screening has evolved from a simple glass-slide smear to a sophisticated test involving liquid-based processing, automated screening, and molecular human papillomavirus (HPV) testing. Recent emphasis on molecular testing seeks to identify HPV strains considered “high-risk” for carcinogenesis, with HPV 16 & 18 being the main focus of research and clinical practice. But is molecular testing more effective and efficient than morphologic testing for cervical screening? Does current data on HPV hold true across all populations? As a public health laboratory serving high-risk, underserved populations, these remain important considerations for our practice. Materials and Methods: Correlation of Pap and HPV results was performed via retrospective review, focusing on Pap cases with high-grade diagnoses and an associated HPV test using the cobasÒ 4800 HPV platform from Roche Diagnostics. HPV results included HPV 16, HPV 18, HPV Other High-Risk, a combination of two or more groups, or HPV Not Detected. The subject population consisted mostly of young women within 200% or less of the poverty line. Results: Of 2,818 cytology test cases reviewed from July 2013-March 2014, 74 were diagnosed as high-grade. HPV was not detected in 10 (13.51%) of these cases. Of the 64 positive HPV tests, 25 (39.06%) fell in the Other High-Risk category. In total, 35 (52.57%) of the high-grade Pap cases were not HPV 16/18 positive. Conclusions: With recent changes to cervical screening practices and guidelines, namely the emphasis on molecular HPV testing, the results of this review are concerning. How will these changes in screening affect our bottom line, both financially and prognostically? Is our patient population substantially different from those used to develop popular testing algorithms? As we move forward with evolution of cervical screening practices, it will be important to explore these questions for the continued quality and integrity of women’s health services.
Figure 1
Figure 2
Cumulative risk of CIN2+ by HPV or Pap cytology testing
Cumulative risk of CIN2+ by HPV and Pap cytology testing
67 Can HPV Testing Be Used as a First-line Screening Test for Cervical Cancer Prevention? Three-year Cumulative Risk of CIN2+ In Women Aged 30 Years and Older Screened by Pap and HPV Co-testing
68
Abha Khanna, MA, CT(ASCP), Marilyn Dawlett, CT(ASCP), Teresa Kologinczak, BS, SCT(ASCP), IAC, Jianping Wang, PhD, CT(ASCP), Shobhana Patel, BS, CT(ASCP), Therese Bevers, MD, Nour Sneige, MD, Ming Guo, MD. University of Texas MD Anderson Cancer Center, Houston, Texas
Aparna Mahajan, MD1, Erek Kucher, BS, CT (ASCP)2, Shawna Engelhardt, CT(ASCP)2, William Rehrauer, PhD3, Wanda Hoefle, MT(ASCP)3, Suzanne Selvaggi, MD3. 1University of Wisconsin Hospital, Madison, Wisconsin; 2University of Wisconsin Hospital and Clinics, Madison, Wisconsin; 3University of Wisconsin School of Medicine and Public Health, Madison, Wisconsin
Introduction: Recently, HPV testing was approved as a first-line screening tool for cervical cancer prevention by the U.S. Food and Drug Administration. To evaluate whether HPV testing can be used effectively as a first-line
Maximizing the Adequacy of CervistaÒ HPV Results on ThinPrepÒ Pap Samples Treated with Glacial Acetic Acid
Introduction: Bloody ThinPrepÒ Pap test samples (TPTS) frequently have high unsatisfactory rates. Since 2002, bloody TPTS have been
Abstracts
S39
prospectively treated with Glacial Acetic Acid (GAA) in our laboratory to lyse blood and improve specimen adequacy. However, studies have shown that treated TPTS have higher rates of insufficient levels of genomic DNA for HologicÒ CervistaÒ HPV HR assay as compared to untreated samples. The aims of this pilot study were to assess the adequacy of the HologicÒ CervistaÒ HPV test results on TPTS prior to GAA treatment, and to determine the effects of pre-aliquoted TPTS on the cytologic unsatisfactory rate. Materials and Methods: Prior to GAA treatment, 2.5 ml of the sample was aliquoted from the 36 bloody TPTS vials. Both pre and post GAA specimens on 36 samples were analyzed using HologicÒ CervistaÒ HPV HR InvaderÒ assay which utilizes the human histone 2 gene (H2be, HIST2H2BE) as an internal control to determine the relative quality of sample genomic DNA. Levels of genomic DNA were compared between these samples. The cytologic unsatisfactory rate of TPTS prospectively treated with GAA was compared to pre-aliquoted samples. Results: Table 1: Comparison of adequacy of genomic DNA and TPTS unsatisfactory rate between pre-aliquoted samples and those treated with GAA Conclusion: Pre-aliquoting samples for HPV testing prior to GAA treatment is an effective method for maximizing the adequacy of the HologicÒ CervistaÒ HPV HR test without significantly impacting TPTS adequacy. The presence of blood does not appear to negatively affect HologicÒ CervistaÒ HPV results. Applying this technique does not significantly impact TPTS unsatisfactory rates. Table 1
Adequate level of genomic DNA TPTS Unsatisfactory rate
Pre-aliquoted sample (untreated with GAA)
Sample treated with GAA
Statistical Significance
100% (36/36)
19.4% (7/36)
p<0.00001 (Significant)
8.3% (3/36)
2.2% (13/581)
pZ0.089 (Not Significant)
69 High-Risk HPV is Associated with Other Vaginal Infections Gabriela Oprea-Ilies, MD1, Geoffrey Smith, MD2, David Martin, MD2, Ifeoma Nwadei, MD2, Marina Mosunjac, MD2, Tadros Tallat2. 1Emory University School of Medicine, Atlanta, Georgia; 2Emory University, Atlanta, Georgia Introduction: ASC-US represents a cytopathologic interpretation of uncertainty. By new Bethesda guidelines, a liquid prep Pap test with this interpretation is reflex tested for the presence of human papilloma virus (HPV). FDA approved the use of HPV testing in conjunction with the liquid prep Pap Test for cervical cancer screening in March 2003. Our institution, which serves a high-risk population, uses a commercial probe to test for high-risk HPV (HR-HPV) in liquid prep Pap tests. We aimed to study the association of vaginal infections such as candida, bacterial vaginosis, trichomonas vaginalis, herpes virus, and actinomyces with the presence of HR-HPV infection. Material and Methods: We searched the files of our institution from 20062009 for Pap tests diagnosed as ASC-US and having HR-HPV test. These cases were studied in correlation with associated vaginal infections. Additional data included age at the time of the test. The data were entered into an excel spreadsheet. Statistical calculation was performed using twotailed Fisher exact test. Results: A total of 6144 Pap tests were diagnosed as ASC-US, of which 5308 had HR-HPV test performed. 1845 were positive and 3463 were negative. The remaining Pap tests have not been tested for HR-HPV. The age distribution was: 1557 (84%) younger than 50 years of age in HPV positive cases and 2622 (76%) in HPV negative cases, representing a statistically significant difference (pZ0.0001). Results of vaginal infections associated with HR-HPV status on liquid Pap test are summarized in Table 1.
Conclusions: 1. Pap test with a morphologically diagnostic infectious organisms is most likely correlated with HR-HPV infection. Table 1
CANDIDA TRICHOMONAS BACTERIAL V. COMBINED INFECTIONS ALL IMNFECTIONS
HR-HPV + NZ 1845 (35%)
HR-HPV e NZ3463 (65%)
P-Value
170 147 332 48 697
280 271 456 %5 1162
0.1627 0.9157 0.0001 0.0122 0.023
(9%) (8%) (19%) (35%) (38%)
(8) (8 %) (13%) (2%) (34%)
2. The presence of overt infectious organisms may be a pitfall for cytopathologists who may think to downgrade the nuclear changes to “reactive” only and fail to test for the presence of HPV. 70 Comparing Histological Outcome for Cytology Negative/HPV+ with ASCUS/HPV e and ASCUS/HPV + Wanwisa Himakhun, MD1, Jianyu Rao, MD2, Po Chu Fung, MBA, SCT(ASCP)2, Minhui Wang, MD, PhD2, Melody Castellon, BS2, Sophia Apple, MD2. 1University of California at Los Angeles Medical Center, Los Angeles, California; 2University of California at Los Angeles, Los Angeles, California Introduction: A major change of 2012 ASCCP guidelines is to include HPV co-testing with cytology in women age 30. It is recommended that for Cytology -/ HPV+, either HPV typing or repeat-co-testing are acceptable. Repeat co-testing in 3 years for ASCUS/HPV- is recommended. To determine the validity of these recommendations, we aimed to evaluate the histology outcomes of subjects with discordant cytology and HPV findings in cytology-negative and ASCUS groups, stratified by age cut-off at 30. Materials and Methods: A retrospective study from January 1, 2011 to 30 June, 2013 was performed on liquid-based cytology that had HrHPV testing (digene HC2). The Cytology -/HPV +, ASCUS/ HPV+ and ASCUS/ HPVgroups were studied using surgical biopsy results as gold standard. Each group was separated into ages Results: From 17,103 cytology-co-testing within the study time period, 1,288 cases met our criteria for study groups. The Cytology-/HPV +, ASCUS/HPV + and ASCUS/HPV- cases were 149, 897 and 242 (Table 1, Figure 1) respectively. The average follow-up time was 6 months. The ASCUS/HPV+ group showed 51% SIL including 11% HSIL. Cytology-/ HPV+ had 39% SIL with 6% HSIL and ASCUS/HPV- had 30% SIL with 2% HSIL (Figure 1). Interestingly, in the Cytology-/HPV+ group, women Conclusions: The findings in general support the 2012 ASCCP management guidelines. However, for Cytology-/HPV+, colposcopy instead of repeat co-testing in 12 months may be considered as there is significant risk for SIL, including 6% CIN2 and above lesions, especially in women age < 30. Likewise, ASCUS/HPV- also carries significant SIL risk and repeat co-testing at 3 years rather than 12 months may miss SIL lesions, including occasional CIN2 and above lesions.
Figure 1.
Abstracts
Graph of Table 2.
66 Molecular Testing and Cervical Screening: Will One Test Fit All? Erin McCarthy, BS, CT(ASCP)1, Changhong Ye, BS, SCT(ASCP)2, Daniel Kurtycz, MD2. 1University of Wisconsin, Madison, Wisconsin; 2 Wisconsin State Laboratory of Hygiene, Madison, Wisconsin
screening tool, we compared the risk levels of high-grade cervical intraepithelial neoplasia (CIN2+) during a 3-year follow-up period in women aged 30 years and older who screened with HPV and Pap co-testing. Materials and Methods: Patient records from the Cancer Prevention Center were searched retrospectively for women aged 30 years and older who underwent cervical cancer screening during 2007 and 2008. Screening consisted of SurePath Pap and Hybrid Capture 2 (HC2) HPV co-testing. Cumulative incidences of CIN2+ were compared to evaluate the risks stratified by HPV and/or Pap testing results. Results: During the study period, 5,739 women underwent screening. HPV was positive in 3.75% (215/5,739) of the women. Of the 2,107 women for whom 3-year follow-up data were available, 189 (9.0%) had a positive HPV result and 188 (8.9%) had a positive Pap result (ASC-US+). During the 3year follow-up period, 20 women had a CIN2+ biopsy result (10 CIN2 cases, 10 CIN3 cases). The cumulative risk of CIN2+ in women with a positive HPV result (8.5%) was significantly higher than that in women with a negative HPV result (0.26%; P Conclusions: During a 3-year screening interval, the risk of CIN2+ in women aged 30 years and older with a HPV-negative result is sufficiently low to ensure a safety margin for cervical cancer prevention, supporting the use of HPV testing as a first-line screening tool for cervical cancer prevention in women aged 30 and older.
Introduction: Over time, cervical screening has evolved from a simple glass-slide smear to a sophisticated test involving liquid-based processing, automated screening, and molecular human papillomavirus (HPV) testing. Recent emphasis on molecular testing seeks to identify HPV strains considered “high-risk” for carcinogenesis, with HPV 16 & 18 being the main focus of research and clinical practice. But is molecular testing more effective and efficient than morphologic testing for cervical screening? Does current data on HPV hold true across all populations? As a public health laboratory serving high-risk, underserved populations, these remain important considerations for our practice. Materials and Methods: Correlation of Pap and HPV results was performed via retrospective review, focusing on Pap cases with high-grade diagnoses and an associated HPV test using the cobasÒ 4800 HPV platform from Roche Diagnostics. HPV results included HPV 16, HPV 18, HPV Other High-Risk, a combination of two or more groups, or HPV Not Detected. The subject population consisted mostly of young women within 200% or less of the poverty line. Results: Of 2,818 cytology test cases reviewed from July 2013-March 2014, 74 were diagnosed as high-grade. HPV was not detected in 10 (13.51%) of these cases. Of the 64 positive HPV tests, 25 (39.06%) fell in the Other High-Risk category. In total, 35 (52.57%) of the high-grade Pap cases were not HPV 16/18 positive. Conclusions: With recent changes to cervical screening practices and guidelines, namely the emphasis on molecular HPV testing, the results of this review are concerning. How will these changes in screening affect our bottom line, both financially and prognostically? Is our patient population substantially different from those used to develop popular testing algorithms? As we move forward with evolution of cervical screening practices, it will be important to explore these questions for the continued quality and integrity of women’s health services.
Figure 1
Figure 2
Cumulative risk of CIN2+ by HPV or Pap cytology testing
Cumulative risk of CIN2+ by HPV and Pap cytology testing
67 Can HPV Testing Be Used as a First-line Screening Test for Cervical Cancer Prevention? Three-year Cumulative Risk of CIN2+ In Women Aged 30 Years and Older Screened by Pap and HPV Co-testing
68
Abha Khanna, MA, CT(ASCP), Marilyn Dawlett, CT(ASCP), Teresa Kologinczak, BS, SCT(ASCP), IAC, Jianping Wang, PhD, CT(ASCP), Shobhana Patel, BS, CT(ASCP), Therese Bevers, MD, Nour Sneige, MD, Ming Guo, MD. University of Texas MD Anderson Cancer Center, Houston, Texas
Aparna Mahajan, MD1, Erek Kucher, BS, CT (ASCP)2, Shawna Engelhardt, CT(ASCP)2, William Rehrauer, PhD3, Wanda Hoefle, MT(ASCP)3, Suzanne Selvaggi, MD3. 1University of Wisconsin Hospital, Madison, Wisconsin; 2University of Wisconsin Hospital and Clinics, Madison, Wisconsin; 3University of Wisconsin School of Medicine and Public Health, Madison, Wisconsin
Introduction: Recently, HPV testing was approved as a first-line screening tool for cervical cancer prevention by the U.S. Food and Drug Administration. To evaluate whether HPV testing can be used effectively as a first-line
Maximizing the Adequacy of CervistaÒ HPV Results on ThinPrepÒ Pap Samples Treated with Glacial Acetic Acid
Introduction: Bloody ThinPrepÒ Pap test samples (TPTS) frequently have high unsatisfactory rates. Since 2002, bloody TPTS have been
Abstracts
S39
prospectively treated with Glacial Acetic Acid (GAA) in our laboratory to lyse blood and improve specimen adequacy. However, studies have shown that treated TPTS have higher rates of insufficient levels of genomic DNA for HologicÒ CervistaÒ HPV HR assay as compared to untreated samples. The aims of this pilot study were to assess the adequacy of the HologicÒ CervistaÒ HPV test results on TPTS prior to GAA treatment, and to determine the effects of pre-aliquoted TPTS on the cytologic unsatisfactory rate. Materials and Methods: Prior to GAA treatment, 2.5 ml of the sample was aliquoted from the 36 bloody TPTS vials. Both pre and post GAA specimens on 36 samples were analyzed using HologicÒ CervistaÒ HPV HR InvaderÒ assay which utilizes the human histone 2 gene (H2be, HIST2H2BE) as an internal control to determine the relative quality of sample genomic DNA. Levels of genomic DNA were compared between these samples. The cytologic unsatisfactory rate of TPTS prospectively treated with GAA was compared to pre-aliquoted samples. Results: Table 1: Comparison of adequacy of genomic DNA and TPTS unsatisfactory rate between pre-aliquoted samples and those treated with GAA Conclusion: Pre-aliquoting samples for HPV testing prior to GAA treatment is an effective method for maximizing the adequacy of the HologicÒ CervistaÒ HPV HR test without significantly impacting TPTS adequacy. The presence of blood does not appear to negatively affect HologicÒ CervistaÒ HPV results. Applying this technique does not significantly impact TPTS unsatisfactory rates. Table 1
Adequate level of genomic DNA TPTS Unsatisfactory rate
Pre-aliquoted sample (untreated with GAA)
Sample treated with GAA
Statistical Significance
100% (36/36)
19.4% (7/36)
p<0.00001 (Significant)
8.3% (3/36)
2.2% (13/581)
pZ0.089 (Not Significant)
69 High-Risk HPV is Associated with Other Vaginal Infections Gabriela Oprea-Ilies, MD1, Geoffrey Smith, MD2, David Martin, MD2, Ifeoma Nwadei, MD2, Marina Mosunjac, MD2, Tadros Tallat2. 1Emory University School of Medicine, Atlanta, Georgia; 2Emory University, Atlanta, Georgia Introduction: ASC-US represents a cytopathologic interpretation of uncertainty. By new Bethesda guidelines, a liquid prep Pap test with this interpretation is reflex tested for the presence of human papilloma virus (HPV). FDA approved the use of HPV testing in conjunction with the liquid prep Pap Test for cervical cancer screening in March 2003. Our institution, which serves a high-risk population, uses a commercial probe to test for high-risk HPV (HR-HPV) in liquid prep Pap tests. We aimed to study the association of vaginal infections such as candida, bacterial vaginosis, trichomonas vaginalis, herpes virus, and actinomyces with the presence of HR-HPV infection. Material and Methods: We searched the files of our institution from 20062009 for Pap tests diagnosed as ASC-US and having HR-HPV test. These cases were studied in correlation with associated vaginal infections. Additional data included age at the time of the test. The data were entered into an excel spreadsheet. Statistical calculation was performed using twotailed Fisher exact test. Results: A total of 6144 Pap tests were diagnosed as ASC-US, of which 5308 had HR-HPV test performed. 1845 were positive and 3463 were negative. The remaining Pap tests have not been tested for HR-HPV. The age distribution was: 1557 (84%) younger than 50 years of age in HPV positive cases and 2622 (76%) in HPV negative cases, representing a statistically significant difference (pZ0.0001). Results of vaginal infections associated with HR-HPV status on liquid Pap test are summarized in Table 1.
Conclusions: 1. Pap test with a morphologically diagnostic infectious organisms is most likely correlated with HR-HPV infection. Table 1
CANDIDA TRICHOMONAS BACTERIAL V. COMBINED INFECTIONS ALL IMNFECTIONS
HR-HPV + NZ 1845 (35%)
HR-HPV e NZ3463 (65%)
P-Value
170 147 332 48 697
280 271 456 %5 1162
0.1627 0.9157 0.0001 0.0122 0.023
(9%) (8%) (19%) (35%) (38%)
(8) (8 %) (13%) (2%) (34%)
2. The presence of overt infectious organisms may be a pitfall for cytopathologists who may think to downgrade the nuclear changes to “reactive” only and fail to test for the presence of HPV. 70 Comparing Histological Outcome for Cytology Negative/HPV+ with ASCUS/HPV e and ASCUS/HPV + Wanwisa Himakhun, MD1, Jianyu Rao, MD2, Po Chu Fung, MBA, SCT(ASCP)2, Minhui Wang, MD, PhD2, Melody Castellon, BS2, Sophia Apple, MD2. 1University of California at Los Angeles Medical Center, Los Angeles, California; 2University of California at Los Angeles, Los Angeles, California Introduction: A major change of 2012 ASCCP guidelines is to include HPV co-testing with cytology in women age 30. It is recommended that for Cytology -/ HPV+, either HPV typing or repeat-co-testing are acceptable. Repeat co-testing in 3 years for ASCUS/HPV- is recommended. To determine the validity of these recommendations, we aimed to evaluate the histology outcomes of subjects with discordant cytology and HPV findings in cytology-negative and ASCUS groups, stratified by age cut-off at 30. Materials and Methods: A retrospective study from January 1, 2011 to 30 June, 2013 was performed on liquid-based cytology that had HrHPV testing (digene HC2). The Cytology -/HPV +, ASCUS/ HPV+ and ASCUS/ HPVgroups were studied using surgical biopsy results as gold standard. Each group was separated into ages Results: From 17,103 cytology-co-testing within the study time period, 1,288 cases met our criteria for study groups. The Cytology-/HPV +, ASCUS/HPV + and ASCUS/HPV- cases were 149, 897 and 242 (Table 1, Figure 1) respectively. The average follow-up time was 6 months. The ASCUS/HPV+ group showed 51% SIL including 11% HSIL. Cytology-/ HPV+ had 39% SIL with 6% HSIL and ASCUS/HPV- had 30% SIL with 2% HSIL (Figure 1). Interestingly, in the Cytology-/HPV+ group, women Conclusions: The findings in general support the 2012 ASCCP management guidelines. However, for Cytology-/HPV+, colposcopy instead of repeat co-testing in 12 months may be considered as there is significant risk for SIL, including 6% CIN2 and above lesions, especially in women age < 30. Likewise, ASCUS/HPV- also carries significant SIL risk and repeat co-testing at 3 years rather than 12 months may miss SIL lesions, including occasional CIN2 and above lesions.
Figure 1.