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Changes of High Risk HPV (HR HPV) Type Distribution from Atypical Squamous Cells of Undetermined Significance (ASCUS) to High-grade Squamous Intraepithelial Lesion (HSIL)
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Abstracts
Conclusion: There appears to be a discrepancy between the screening assays for the AptimaÒ HPV High Risk Screen as compared to the AptimaÒ HPV High Risk Genotyping assay. The disadvantage of the Aptima Ò assay, as compared to the RocheÒ assay, is that it is not run as an adjunctive test and requires a reflex of the High Risk screen to the Genotyping assay. The screening assay did not detect a small subset (1.9%) of the type 16 HPV genotype. The screening assay appears less sensitive than the genotyping assay for the type 16 HPV.
97% of women with a HSIL pap result test positive for high-risk HPV. The overexpression of HPV E6 and E7 oncogenes are associated with cell proliferation and cervical carcinogenesis. In this study, we demonstrate that overexpression of these mRNAs can be used as a possible progression marker for HSIL. Materials and Methods: A total of 87 HSIL samples, positive for high-risk type HPV (AptimaÒ HPV Assay), were obtained from a pool of 3,621 liquid-based samples (ThinPrep and SurePath Pap Test) from OB/GYN clinics around Puerto Rico. Samples were rescreened and classified following TBS criteria for HSIL. Age range was 17 to 65 years old. To determine overexpression of HPV E6/E7 mRNA, each sample was submitted to OncoProbe assay test (IncellDxTM) and was classified as positive in cases where >2% of cells showed overexpression of E6/E7 mRNA. Analysis for predicting association between overexpression of E6/E7 mRNA and the risk of progression in HSIL cases was done. Results: 87 HSIL cytology samples: 39 cases (45%) showed overexpression of E6/E7 mRNA and were classified as positive; 48 cases (55%) were classified as negative. Association between the risk of progression (40-50%) and the overexpression of E6/E7 mRNA in HSIL cases (45%) was identified. Conclusion: This study demonstrated that overexpression of E6/E7 mRNA is a powerful tool that could be used to determine the risk of progression in HSIL cases in a Puerto Rican population. 84
% HPV Positive Genotypes With Negative HR Screen
Changes of High Risk HPV (HR HPV) Type Distribution from Atypical Squamous Cells of Undetermined Significance (ASCUS) to High-grade Squamous Intraepithelial Lesion (HSIL) Pradip Manna, PhD, MBA, Spencer Kerley, MD, Paula Clark, BS, MT(ASCP), George Ryan, BS, Greg Scholtz, MS, Joseph Kerley, BA, Stacey Golub, BS, Kathy Goudy, BA, CT(ASCP)IAC. Physicians Reference Laboratory, Overland Park, Kansas
% NILM and LSIL Cases With Positive Type 16
83 Overexpression of HPV E6/E7 mRNA: A Possible Progression Marker for High-grade Squamous Intraepithelial Lesions in a Puerto Rican Population Katian Melendez, CT(ASCP), Lilliam Echevarria, Ivan Garcia. Dr. Noy Pathology Laboratory, Luquillo, Puerto Rico Introduction: High-grade squamous intraepithelial lesions (HSIL) constitute about 0.5% of all cervical and vaginal samples. According to The Bethesda System (TBS), HSIL criteria include changes in less mature cells showing a marked increase in the nuclear/cytoplasmic ratios, irregular nuclear membranes and nuclear hyperchromasia. HSIL carries a higher risk of progression to cervical cancer given that nearly 90% of LSIL do not progress, and 50% to 60% of HSIL regress. Approximately,
Introduction: Understanding HR HPV type distribution is essential to cervical cancer management. Particular types of HPV infection may preferentially progress from ASCUS to HSIL and then to invasive carcinoma. Genotyping improves risk stratification of HR HPV-positive women in cervical screening programs. We used Complete Care HPV, a next generation HPV test that simultaneously detects, types, and quantifies all 15 HR HPV types to determine type distribution in ASCUS and HSIL cases in the Kansas City region. Materials and Methods: We evaluated 15 HR HPV type distributions of 7,320 women with ASCUS and 168 women with HSIL cytologies. The majority of the specimens were ThinPrep Pap. Results: Forty-two percent of the ASCUS and 98% of the HSIL were positive for HR HPV (Table 1). Of the HPV positive ASCUS and HSIL cases, 39% and 51% had multiple infections. Both cases had similar prevalence rates of double, triple, quadruple, quintuple, and sextuple infections. Prevalence rates of 15 HR HPV types were 4-28% in ASCUS and 3-63% in HSIL (Table 2). HPV16 was most prevalent with 28% in ASCUS and 63% in HSIL. HPV18 was the eighth and sixth most prevalent in ASCUS and HSIL, but was less prevalent than HPV31, 39, 51, 52, 56, and 59. Individual HPV types were more prevalent in multiple infections than their corresponding single infections (Table 3, Table 4). Conclusions: Significant changes in the HR HPV type distribution were observed with the progression of cytological abnormalities from ASCUS to HSIL. Notably, HPV16 increased from 13% to 29% in single and from 15% to 34% in multiple infections from ASCUS to HSIL. With nearly half of the HSIL cases, multiple infections clearly played a critical role in cervical cancer. Both multiple infections and the somewhat-diminished role of HPV18 may impact the future of cervical cancer management through vaccination.
Abstracts
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85 Diagnostic Utility of p16 and Ki-67 Immunostains on Cell Blocks from Papanicolaou Tests (Liquid-based Preparation) Fan Lin, MD, PhD, Asif Shahab, MD, Haiyan Liu, MD. Geisinger Medical Center, Danville, Pennsylvania Introduction: The Papanicolaou (Pap) test is a widely accepted screening test for cervical cancer. Although the Pap test is considered an excellent screening modality, it also poses diagnostic challenges in certain cases. Cell block (CB) preparation has been routinely utilized in non-gynecological specimens with proven value; however, it is not routinely used in conjunction with Pap tests. In this study, we retrospectively reviewed Pap tests with corresponding CBs, immunostains, and follow-up data to evaluate the diagnostic utility of CBs in conjunction with immunostains in Pap tests. Materials and Methods: One hundred forty-two liquid-based Pap smears with corresponding CBs and immunostains (p16 and Ki-67) from the archives of Laboratory Medicine at Geisinger Medical Center from 2006 to 2012 were retrieved and reviewed. Available follow-up data were analyzed. Results: Overall 94% (133/142) of the cases had adequate CB material for immunostains and contributed to the final diagnosis. Seventy-two percent (103/142) had follow-up data and were further analyzed as summarized in Table 1. In the squamous intraepithelial lesion (SIL) group, 100% (6/6) were diagnosed as high-grade dysplasia on follow-up. Among the atypical squamous cells, cannot rule out high-grade squamous intraepithelial lesion (ASC-H) cases, 67% (10/15) confirmed dysplasia. Of the 16 atypical glandular cells of undetermined significance (AGUS) cases, 44% (7/16) were confirmed to be dysplasia or were positive for human papilloma virus (HPV) on follow-up. Of the 60 benign cases with follow-up data, 98% (59/ 60) continued to be benign. Positive immunostaining for p16 and an increased Ki-67 labeling index were seen in all cases of abnormal Pap smears. Conclusions: Our study demonstrated that the majority of cases yielded adequate cellular material in CB for further immunostains and contributed to the final diagnosis. These data support the notion of utilizing CBs in conjunction with immunostains as a useful adjunct to challenging Pap smears.
Abstracts
Conclusion: There appears to be a discrepancy between the screening assays for the AptimaÒ HPV High Risk Screen as compared to the AptimaÒ HPV High Risk Genotyping assay. The disadvantage of the Aptima Ò assay, as compared to the RocheÒ assay, is that it is not run as an adjunctive test and requires a reflex of the High Risk screen to the Genotyping assay. The screening assay did not detect a small subset (1.9%) of the type 16 HPV genotype. The screening assay appears less sensitive than the genotyping assay for the type 16 HPV.
97% of women with a HSIL pap result test positive for high-risk HPV. The overexpression of HPV E6 and E7 oncogenes are associated with cell proliferation and cervical carcinogenesis. In this study, we demonstrate that overexpression of these mRNAs can be used as a possible progression marker for HSIL. Materials and Methods: A total of 87 HSIL samples, positive for high-risk type HPV (AptimaÒ HPV Assay), were obtained from a pool of 3,621 liquid-based samples (ThinPrep and SurePath Pap Test) from OB/GYN clinics around Puerto Rico. Samples were rescreened and classified following TBS criteria for HSIL. Age range was 17 to 65 years old. To determine overexpression of HPV E6/E7 mRNA, each sample was submitted to OncoProbe assay test (IncellDxTM) and was classified as positive in cases where >2% of cells showed overexpression of E6/E7 mRNA. Analysis for predicting association between overexpression of E6/E7 mRNA and the risk of progression in HSIL cases was done. Results: 87 HSIL cytology samples: 39 cases (45%) showed overexpression of E6/E7 mRNA and were classified as positive; 48 cases (55%) were classified as negative. Association between the risk of progression (40-50%) and the overexpression of E6/E7 mRNA in HSIL cases (45%) was identified. Conclusion: This study demonstrated that overexpression of E6/E7 mRNA is a powerful tool that could be used to determine the risk of progression in HSIL cases in a Puerto Rican population. 84
% HPV Positive Genotypes With Negative HR Screen
Changes of High Risk HPV (HR HPV) Type Distribution from Atypical Squamous Cells of Undetermined Significance (ASCUS) to High-grade Squamous Intraepithelial Lesion (HSIL) Pradip Manna, PhD, MBA, Spencer Kerley, MD, Paula Clark, BS, MT(ASCP), George Ryan, BS, Greg Scholtz, MS, Joseph Kerley, BA, Stacey Golub, BS, Kathy Goudy, BA, CT(ASCP)IAC. Physicians Reference Laboratory, Overland Park, Kansas
% NILM and LSIL Cases With Positive Type 16
83 Overexpression of HPV E6/E7 mRNA: A Possible Progression Marker for High-grade Squamous Intraepithelial Lesions in a Puerto Rican Population Katian Melendez, CT(ASCP), Lilliam Echevarria, Ivan Garcia. Dr. Noy Pathology Laboratory, Luquillo, Puerto Rico Introduction: High-grade squamous intraepithelial lesions (HSIL) constitute about 0.5% of all cervical and vaginal samples. According to The Bethesda System (TBS), HSIL criteria include changes in less mature cells showing a marked increase in the nuclear/cytoplasmic ratios, irregular nuclear membranes and nuclear hyperchromasia. HSIL carries a higher risk of progression to cervical cancer given that nearly 90% of LSIL do not progress, and 50% to 60% of HSIL regress. Approximately,
Introduction: Understanding HR HPV type distribution is essential to cervical cancer management. Particular types of HPV infection may preferentially progress from ASCUS to HSIL and then to invasive carcinoma. Genotyping improves risk stratification of HR HPV-positive women in cervical screening programs. We used Complete Care HPV, a next generation HPV test that simultaneously detects, types, and quantifies all 15 HR HPV types to determine type distribution in ASCUS and HSIL cases in the Kansas City region. Materials and Methods: We evaluated 15 HR HPV type distributions of 7,320 women with ASCUS and 168 women with HSIL cytologies. The majority of the specimens were ThinPrep Pap. Results: Forty-two percent of the ASCUS and 98% of the HSIL were positive for HR HPV (Table 1). Of the HPV positive ASCUS and HSIL cases, 39% and 51% had multiple infections. Both cases had similar prevalence rates of double, triple, quadruple, quintuple, and sextuple infections. Prevalence rates of 15 HR HPV types were 4-28% in ASCUS and 3-63% in HSIL (Table 2). HPV16 was most prevalent with 28% in ASCUS and 63% in HSIL. HPV18 was the eighth and sixth most prevalent in ASCUS and HSIL, but was less prevalent than HPV31, 39, 51, 52, 56, and 59. Individual HPV types were more prevalent in multiple infections than their corresponding single infections (Table 3, Table 4). Conclusions: Significant changes in the HR HPV type distribution were observed with the progression of cytological abnormalities from ASCUS to HSIL. Notably, HPV16 increased from 13% to 29% in single and from 15% to 34% in multiple infections from ASCUS to HSIL. With nearly half of the HSIL cases, multiple infections clearly played a critical role in cervical cancer. Both multiple infections and the somewhat-diminished role of HPV18 may impact the future of cervical cancer management through vaccination.
Abstracts
S45
85 Diagnostic Utility of p16 and Ki-67 Immunostains on Cell Blocks from Papanicolaou Tests (Liquid-based Preparation) Fan Lin, MD, PhD, Asif Shahab, MD, Haiyan Liu, MD. Geisinger Medical Center, Danville, Pennsylvania Introduction: The Papanicolaou (Pap) test is a widely accepted screening test for cervical cancer. Although the Pap test is considered an excellent screening modality, it also poses diagnostic challenges in certain cases. Cell block (CB) preparation has been routinely utilized in non-gynecological specimens with proven value; however, it is not routinely used in conjunction with Pap tests. In this study, we retrospectively reviewed Pap tests with corresponding CBs, immunostains, and follow-up data to evaluate the diagnostic utility of CBs in conjunction with immunostains in Pap tests. Materials and Methods: One hundred forty-two liquid-based Pap smears with corresponding CBs and immunostains (p16 and Ki-67) from the archives of Laboratory Medicine at Geisinger Medical Center from 2006 to 2012 were retrieved and reviewed. Available follow-up data were analyzed. Results: Overall 94% (133/142) of the cases had adequate CB material for immunostains and contributed to the final diagnosis. Seventy-two percent (103/142) had follow-up data and were further analyzed as summarized in Table 1. In the squamous intraepithelial lesion (SIL) group, 100% (6/6) were diagnosed as high-grade dysplasia on follow-up. Among the atypical squamous cells, cannot rule out high-grade squamous intraepithelial lesion (ASC-H) cases, 67% (10/15) confirmed dysplasia. Of the 16 atypical glandular cells of undetermined significance (AGUS) cases, 44% (7/16) were confirmed to be dysplasia or were positive for human papilloma virus (HPV) on follow-up. Of the 60 benign cases with follow-up data, 98% (59/ 60) continued to be benign. Positive immunostaining for p16 and an increased Ki-67 labeling index were seen in all cases of abnormal Pap smears. Conclusions: Our study demonstrated that the majority of cases yielded adequate cellular material in CB for further immunostains and contributed to the final diagnosis. These data support the notion of utilizing CBs in conjunction with immunostains as a useful adjunct to challenging Pap smears.