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Patterns and Reliability of p16 Immunohistochemistry (IHC) in Cytology Specimens
Abstracts chosen to reflect an atypical rate similar to our normal practice: 199 NILM, 31 ASCUS, and 20 LSIL or worse. Each slide was independently reevaluated by 8 cytotechnologists blinded to the original diagnosis and study purpose. The study coordinator was aware of the HPV status in all cases; 50% of all selected NILM and ASCUS cases were HPV-positive. HPV status was provided to the cytotechnologists in 45% of cases from each HPV subgroup, and withheld in 55%. Results: There were 1997 unique observations: 1597 from NILM cases, 248 from ASCUS cases, and 152 from LSIL or worse cases. There was no difference in upgrading NILM cases to ASCUS or worse for HPV-positive and HPV-negative cases when the HPV status was withheld (27% vs. 29%, pZ0.54). There was significant bias for HPV-positive NILM cases to be upgraded to ASCUS or worse when the HPV-positive status was provided (45% vs. 27%, p Conclusions: Knowledge of HPV status biases cytotechnologist interpretation of NILM cases, but not ASCUS cases. Knowledge of HPV-positive status results in a higher upgrade rate, whereas a known HPV-negative status has the opposite effect. A repeat of the study is underway to assess individual intra-observer variability.
S5 Materials and Methods: The Pap volume for our laboratory is 115,000+ tests annually. In June of 2014, our lab transitioned to the Roche cobasÒ HPV platform. During the period from June 2014-April 2015, we processed and diagnosed 62,700 ThinPrepÒ Paps that had HPV testing performed. We searched our database to identify all HPV negative results with significant cytologic diagnoses of ASC-H or higher from these cases. We included only tests for women 25 years of age with HPV performed on the cobas platform. There were 72 cases with results that fell into these parameters and 52 had histologic follow-up. Biopsy results were correlated with cytologic diagnoses. Results: Of the 52 cases with histologic follow-up, 12 had CIN2-CIN3, 4 had endometrial adenocarcinoma and 1 had a cervical small cell neuroendocrine carcinoma which was uniformly positive for p16 (Table 1). There were no invasive cervical squamous cell carcinomas or cervical adenocarcinomas identified. Conclusion: According to our study, of the women >25 years of age with a diagnosis of ASC-H or higher on a ThinPrepÒ Pap test and a negative Roche cobasÒHPV test, 35 cases showed no significant lesions and 17 cases showed CIN2 or greater on histologic follow-up. It is important to emphasize that the Pap test is a screening test designed to identify cervical squamous lesions. In practice, however, other malignancies may be detected.
PP11 Patterns and Reliability of p16 Immunohistochemistry (IHC) in Cytology Specimens Renu Virk, MD, Stefan Pambuccian, MD, Eva Wojcik, MD, MIAC, Guliz Barkan, MD, Swati Mehrotra, MD Loyola University Medical Center, Maywood, Illinois
PP10 Histologic Follow-up of Women >25 Years of Age with a Cervical ThinPrepÒ Pap Diagnosis of ASC-H or Higher and a Negative Roche cobasÒ HPV DNA Test Teresa Kologinczak, BS, SCT(ASCP), IAC, Stephanie Hamilton, EdD, SCT, MB(ASCP), Ronald Tickman, MD, Nuria Perez-Reyes, MD CellNetix Pathology and Laboratories, Seattle, Washington Introduction: In April 2014, the FDA approved the Roche cobasÒ HPV DNA test for primary cervical cancer screening. Studies have suggested that patients with significant lesions may be misdiagnosed and inadequately triaged with the primary HPV screening algorithm. Some of this data has been reported using the Digene Hybrid Capture 2 DNA test (Qiagen). We report our experience with the cobas assay, comparing HPV negative cases from women 25 years of age, with cytologic and histologic results.
Introduction: p16 immunohistochemistry(IHC) is frequently used as a surrogate marker for oropharyngeal HPV-associated squamous cell carcinoma (SCC) with diffuse block-like nuclear and cytoplasmic p16 staining >70% of tumor cells interpreted as positive. However, the criteria to interpret p16 IHC in cytology specimens (direct smears and cell block sections) are lacking. We conducted this study to assess the pattern and reliability of p16 IHC in fine needle aspiration(FNA) material (cell block sections) obtained from SCC at metastatic sites. Methods and Materials: We analysed patterns of p16 IHC expression and correlated it with the site of primary tumor (oropharyngeal vs. non oropharyngeal) determined based on either additional pathology and/or imaging/clinical studies. We also correlated p16 IHC results on cytology with p16 IHC performed on the surgical material from the primary tumor. The cytology slides were reviewed only for cases where p16 IHC was reported positive. Results: The study included FNAB from neck masses in 31 patients (25 men and 6 women) with mean age of 58 years. An additional 5 cases included other metastatic sites including mediastinal lymph nodes (3), liver (1) and lung (1). Tables 1 and 2 show p16 IHC results with primary site of tumor and patterns of expression in FNA material. Overall, p16 IHC was positive in 21 cases and negative in 13 cases while non-contributory in 2 cases. The sensitivity and specificity of p16 IHC in predicting primary
S6 oropharyngeal SCC were calculated to be 95% and 82% respectively whereas positive and negative predictive value was 90% for each. Conclusion: When strict interpretative criteria are used (>50% of tumor cells with nuclear and cytoplasmic staining), p16 IHC results on FNA cell block sections are reliable and help in localizing the primary site of origin (oropharyngeal vs non-oropharyngeal) with high sensitivity and positive predictive value.
Abstracts Results: 168 cases (62%) were adequate specimens after reprocessing, yielding 163 negative for intraepithelial lesion (NILM) (97%), 4 atypical squamous cells of undetermined significance (ASCUS) (2.4%) and 1 low grade lesion (0.6%). 105 unsatisfactory cases were due to blood (43%), lubricant (20%), scant cellularity (29.5%), inflammation (4.8%), and blood/ lubricant (3%). Follow-up repeat cervical cytology for unsatisfactory specimens in 78 of 102 women demonstrated 12 unsatisfactory specimens, 59 NILMs, 6 ASCUS, and 1 low-grade lesion. Conclusion: SurePath technique utilizes a density reagent which acts as an effective filter to eliminate obscuring factors such as blood, lubricant, and inflammation, without increasing our unsatisfactory rates. With ThinPrep, the unsatisfactory rate is much higher. We processed ThinPrep specimens using SurePath methodology after the methanol fixative was removed. Well preserved cellular morphology and enriched cellularity was achieved. Reprocessing unsatisfactory ThinPrep with the SurePath density reagent is an effective way to decrease the unsatisfactory rate in cervical cytology specimens. EDUCATION/TRAINING/CURRENT TRENDS 1 E-learning Module on Fine Needle Aspiration Technique Maheswari Mukherjee, PhD, MS, CT(ASCP), Amber Donnelly, PhD, MPH, SCT(ASCP), Stanley Radio, MD University of Nebraska Medical Center, Lincoln, Nebraska
PP12 Reprocessing Unsatisfactory ThinPrep Specimens with SurePath Density Reagent Decreases the Unsatisfactory Rate Dianne Grunes, MD, Fang Zhou, MD, Paul Elgert, CT(ASCP)IAC, Aylin Simsir, MD, Melissa Yee-Chang, DO New York University Medical Center, New York, New York Introduction: ThinPrep and SurePath are two widely used liquid-based cervical cytology preparation techniques. SurePath has a lower unsatisfactory rate, but is not approved by the FDA for human papillomavirus testing. For this reason, our laboratory switched to ThinPrep and had a markedly increased unsatisfactory rate (up to 7%) due to lubricants, blood/inflammation, and insufficient cellularity. Our SurePath unsatisfactory rate was less than 1%. Our patient population has limited access to care and restricted flexibility in scheduling gynecologic appointments. Glacial acetic acid treatment for bloody specimens is time-consuming and repeat Thinprep slides did not yield significant improvement. Materials and Methods: The residual samples from 270 specimens initially deemed unsatisfactory were reprocessed by removing methanol fixative with washes of Hank’s balanced salt solution via centrifugation and resuspension in SurePath preservative. Specimens were then processed according to standard SurePath methodology and demonstrated preserved cellular morphology.
Introduction: E-learning is becoming popular among many healthcare educational programs. This study reports our cytotechnology (CT) program students’ perception on a fine needle aspiration (FNA) technique E-learning module. Materials and Methods: Articulate Presenter ‘09 was used to develop an Elearning module that consisted of a) video clips on: overview of an FNA procedure, demonstration of patient interaction, preparation and staining of the glass slide specimens, and evaluation of specimen adequacy; and b) self-assessments. Eight CT students (campus & satellite sites) viewed the FNA E-learning module prior to their classroom lecture on FNA technique. The next day, all the students (nZ8) participated in a small group discussion on the FNA technique in the classroom. During the discussion, the students were encouraged to discuss about the: indications, advantages, and disadvantages of a FNA procedure; role of a pathologist and a cytotechnologist in FNA procedure; different steps involved in specimen smearing and staining techniques; and the role of a cytotechnologist in interpreting specimen adequacy. The following day, all eight students participated in an online survey to report their opinion on the E-learning module. Results: All eight students completely understood the content of the Elearning module. The E-learning module helped the students (nZ8) to better prepare themselves for their role in a FNA procedure. Viewing the E-learning module prior to the classroom lecture gave the students (nZ8) the knowledge about FNA so that they could interact/participate during the classroom lecture and ask for any clarification. Table 1 shows the students’ (nZ8) preferences of using E-learning modules in future. Conclusion: The E-learning module on FNA technique was well accepted by our CT students and may be a valuable tool in implementing the “flipped classroom” approach to CT education.
Table 1
S5 Materials and Methods: The Pap volume for our laboratory is 115,000+ tests annually. In June of 2014, our lab transitioned to the Roche cobasÒ HPV platform. During the period from June 2014-April 2015, we processed and diagnosed 62,700 ThinPrepÒ Paps that had HPV testing performed. We searched our database to identify all HPV negative results with significant cytologic diagnoses of ASC-H or higher from these cases. We included only tests for women 25 years of age with HPV performed on the cobas platform. There were 72 cases with results that fell into these parameters and 52 had histologic follow-up. Biopsy results were correlated with cytologic diagnoses. Results: Of the 52 cases with histologic follow-up, 12 had CIN2-CIN3, 4 had endometrial adenocarcinoma and 1 had a cervical small cell neuroendocrine carcinoma which was uniformly positive for p16 (Table 1). There were no invasive cervical squamous cell carcinomas or cervical adenocarcinomas identified. Conclusion: According to our study, of the women >25 years of age with a diagnosis of ASC-H or higher on a ThinPrepÒ Pap test and a negative Roche cobasÒHPV test, 35 cases showed no significant lesions and 17 cases showed CIN2 or greater on histologic follow-up. It is important to emphasize that the Pap test is a screening test designed to identify cervical squamous lesions. In practice, however, other malignancies may be detected.
PP11 Patterns and Reliability of p16 Immunohistochemistry (IHC) in Cytology Specimens Renu Virk, MD, Stefan Pambuccian, MD, Eva Wojcik, MD, MIAC, Guliz Barkan, MD, Swati Mehrotra, MD Loyola University Medical Center, Maywood, Illinois
PP10 Histologic Follow-up of Women >25 Years of Age with a Cervical ThinPrepÒ Pap Diagnosis of ASC-H or Higher and a Negative Roche cobasÒ HPV DNA Test Teresa Kologinczak, BS, SCT(ASCP), IAC, Stephanie Hamilton, EdD, SCT, MB(ASCP), Ronald Tickman, MD, Nuria Perez-Reyes, MD CellNetix Pathology and Laboratories, Seattle, Washington Introduction: In April 2014, the FDA approved the Roche cobasÒ HPV DNA test for primary cervical cancer screening. Studies have suggested that patients with significant lesions may be misdiagnosed and inadequately triaged with the primary HPV screening algorithm. Some of this data has been reported using the Digene Hybrid Capture 2 DNA test (Qiagen). We report our experience with the cobas assay, comparing HPV negative cases from women 25 years of age, with cytologic and histologic results.
Introduction: p16 immunohistochemistry(IHC) is frequently used as a surrogate marker for oropharyngeal HPV-associated squamous cell carcinoma (SCC) with diffuse block-like nuclear and cytoplasmic p16 staining >70% of tumor cells interpreted as positive. However, the criteria to interpret p16 IHC in cytology specimens (direct smears and cell block sections) are lacking. We conducted this study to assess the pattern and reliability of p16 IHC in fine needle aspiration(FNA) material (cell block sections) obtained from SCC at metastatic sites. Methods and Materials: We analysed patterns of p16 IHC expression and correlated it with the site of primary tumor (oropharyngeal vs. non oropharyngeal) determined based on either additional pathology and/or imaging/clinical studies. We also correlated p16 IHC results on cytology with p16 IHC performed on the surgical material from the primary tumor. The cytology slides were reviewed only for cases where p16 IHC was reported positive. Results: The study included FNAB from neck masses in 31 patients (25 men and 6 women) with mean age of 58 years. An additional 5 cases included other metastatic sites including mediastinal lymph nodes (3), liver (1) and lung (1). Tables 1 and 2 show p16 IHC results with primary site of tumor and patterns of expression in FNA material. Overall, p16 IHC was positive in 21 cases and negative in 13 cases while non-contributory in 2 cases. The sensitivity and specificity of p16 IHC in predicting primary
S6 oropharyngeal SCC were calculated to be 95% and 82% respectively whereas positive and negative predictive value was 90% for each. Conclusion: When strict interpretative criteria are used (>50% of tumor cells with nuclear and cytoplasmic staining), p16 IHC results on FNA cell block sections are reliable and help in localizing the primary site of origin (oropharyngeal vs non-oropharyngeal) with high sensitivity and positive predictive value.
Abstracts Results: 168 cases (62%) were adequate specimens after reprocessing, yielding 163 negative for intraepithelial lesion (NILM) (97%), 4 atypical squamous cells of undetermined significance (ASCUS) (2.4%) and 1 low grade lesion (0.6%). 105 unsatisfactory cases were due to blood (43%), lubricant (20%), scant cellularity (29.5%), inflammation (4.8%), and blood/ lubricant (3%). Follow-up repeat cervical cytology for unsatisfactory specimens in 78 of 102 women demonstrated 12 unsatisfactory specimens, 59 NILMs, 6 ASCUS, and 1 low-grade lesion. Conclusion: SurePath technique utilizes a density reagent which acts as an effective filter to eliminate obscuring factors such as blood, lubricant, and inflammation, without increasing our unsatisfactory rates. With ThinPrep, the unsatisfactory rate is much higher. We processed ThinPrep specimens using SurePath methodology after the methanol fixative was removed. Well preserved cellular morphology and enriched cellularity was achieved. Reprocessing unsatisfactory ThinPrep with the SurePath density reagent is an effective way to decrease the unsatisfactory rate in cervical cytology specimens. EDUCATION/TRAINING/CURRENT TRENDS 1 E-learning Module on Fine Needle Aspiration Technique Maheswari Mukherjee, PhD, MS, CT(ASCP), Amber Donnelly, PhD, MPH, SCT(ASCP), Stanley Radio, MD University of Nebraska Medical Center, Lincoln, Nebraska
PP12 Reprocessing Unsatisfactory ThinPrep Specimens with SurePath Density Reagent Decreases the Unsatisfactory Rate Dianne Grunes, MD, Fang Zhou, MD, Paul Elgert, CT(ASCP)IAC, Aylin Simsir, MD, Melissa Yee-Chang, DO New York University Medical Center, New York, New York Introduction: ThinPrep and SurePath are two widely used liquid-based cervical cytology preparation techniques. SurePath has a lower unsatisfactory rate, but is not approved by the FDA for human papillomavirus testing. For this reason, our laboratory switched to ThinPrep and had a markedly increased unsatisfactory rate (up to 7%) due to lubricants, blood/inflammation, and insufficient cellularity. Our SurePath unsatisfactory rate was less than 1%. Our patient population has limited access to care and restricted flexibility in scheduling gynecologic appointments. Glacial acetic acid treatment for bloody specimens is time-consuming and repeat Thinprep slides did not yield significant improvement. Materials and Methods: The residual samples from 270 specimens initially deemed unsatisfactory were reprocessed by removing methanol fixative with washes of Hank’s balanced salt solution via centrifugation and resuspension in SurePath preservative. Specimens were then processed according to standard SurePath methodology and demonstrated preserved cellular morphology.
Introduction: E-learning is becoming popular among many healthcare educational programs. This study reports our cytotechnology (CT) program students’ perception on a fine needle aspiration (FNA) technique E-learning module. Materials and Methods: Articulate Presenter ‘09 was used to develop an Elearning module that consisted of a) video clips on: overview of an FNA procedure, demonstration of patient interaction, preparation and staining of the glass slide specimens, and evaluation of specimen adequacy; and b) self-assessments. Eight CT students (campus & satellite sites) viewed the FNA E-learning module prior to their classroom lecture on FNA technique. The next day, all the students (nZ8) participated in a small group discussion on the FNA technique in the classroom. During the discussion, the students were encouraged to discuss about the: indications, advantages, and disadvantages of a FNA procedure; role of a pathologist and a cytotechnologist in FNA procedure; different steps involved in specimen smearing and staining techniques; and the role of a cytotechnologist in interpreting specimen adequacy. The following day, all eight students participated in an online survey to report their opinion on the E-learning module. Results: All eight students completely understood the content of the Elearning module. The E-learning module helped the students (nZ8) to better prepare themselves for their role in a FNA procedure. Viewing the E-learning module prior to the classroom lecture gave the students (nZ8) the knowledge about FNA so that they could interact/participate during the classroom lecture and ask for any clarification. Table 1 shows the students’ (nZ8) preferences of using E-learning modules in future. Conclusion: The E-learning module on FNA technique was well accepted by our CT students and may be a valuable tool in implementing the “flipped classroom” approach to CT education.
Table 1