Assessment of Fine Needle Aspiration Specimen Adequacy for High-Risk HPV Detection and Genotyping in Oropharyngeal Squamous Cell Carcinoma

Abstract

Objective: The aim of this study was to determine the adequacy of archived and fresh fine needle aspiration (FNA) specimens from metastatic head and neck squamous cell carcinoma (SCC) for the molecular detection and genotyping of high-risk (HR) HPV. Study Design: Thirty-seven specimens from 26 patients diagnosed by FNA with metastatic SCC were included as retrospective specimens [19 slides stained with Papanicolaou (Pap) and 18 with Diff-Quik® (DQ)]. Twenty fresh FNA specimens from 18 patients were included as prospective specimens. These specimens were analyzed using the standard protocol for ThinPrep® cervical specimens, with a Cervista HR HPV detection kit. The positive specimens were tested for the HPV 16 and 18 genotypes. Results: Forty-four of 57 specimens (77%) had sufficient cells to yield a valid HPV result. The adequacy rate for Pap-stained slides was 15/19 (79%), for DQ-stained slides it was 13/18 (72%), and for fresh needle aspirates it was 16/20 (80%). HR HPV was detected in 23/44 (52%) specimens. Among the 23 HPV-positive specimens, 19 were genotyped as HPV 16 and 1 as HPV 18. Conclusions: HR HPV detection and genotyping can be performed on FNA specimens of head and neck SCC prospectively collected in PreservCyt as well as on archival slides with either Pap or DQ stain.

Overview of Nongynecological Samples Prepared with Liquid-Based Cytology Medium

“…This method requires minimal specimen preparation, eliminates the need to process limited specimens as cell blocks and delivers a quantitative result based on a standard threshold that allows for unambiguous scoring of samples as positive or negative. A few feasibility studies have pointed to the effectiveness of these assays in cytological samples of HNSCCs [26,27,28]. …”

Role of Cytology in the Diagnosis and Management of HPV-Associated Head and Neck Carcinoma

“…Recently, detection methods previously validated for HPV cervical Pap testing including proprietary commercial methods such as Hybrid Capture II [82] and Cervista [83] have successfully detected HPV DNA in HNSCC using archival air-dried, alcohol-fixed, and liquid-based preparations. The most rigorous testing to date has been a feasibility test using Hybrid Capture II on cytologic material obtained from surgically resected HPV-associated HNSCC [82].…”

Performance of Aptima E6/E7 mRNA HPV assays on fine needle aspirates from cervical lymph nodes of patients with metastatic oropharyngeal squamous cell carcinoma

“…Performing an assay for the presence of β-actin cellular DNA and examining the Cervista controls for low genomic DNA or a recorded "invalid" result in the cobas HPV DNA test identified 6 FNA samples as inadequate (Table 4). Adequacy of archived and fresh FNA samples for molecular testing has ranged from 44-87% in previous studies using the internal control of the Cervista HPV DNA assay [19][20][21]. The presence of HR HPV mRNA and/ or DNA or sufficient β-actin DNA in fresh FNA samples in our study showed an adequacy rate of 92%.…”

“…Both LDT's and commercial assays for the detection of HR-HPV DNA have been used on FNA samples. An LDT PCR has used MY09/11 primers [16], whereas the commercial DNA assays used Hybrid Capture (HC2, Qiagen) [17,18]; Cervista (Hologic) [19][20][21]; cobas PCR (Roche) [22,23] and the PCR genotyping INNO-LIPA Extra System Kit (Innogenetics) [24]. Accuracy of HPV testing of FNA compared results to the presence of p16 and HR HPV or HPV16 DNA in tumors using ISH.…”