Validation of ThermoFisher's Papspin for human papillomavirus detection in cervicovaginal specimens using PCR with GP5+/GP6+ primers and the Hybrid Capture II assay

Abstract

The present study aimed to validate ThermoFisher's (Thermo Fisher Scientific, Runcorn, Cheshire, UK) Papspin (PS) for human papillomavirus (HPV) testing by in-house PCR and by the Hybrid Capture II (HC2) assay and to compare the results with those obtained using Specimen Transport Medium (STM) (Digene Diagnostics, Gaithersburg, MD, USA). Forty-five patients underwent conization for known lesions ranging from atypical squamous cells of undetermined significance (ASC-US) with high-risk HPV (hr-HPV) to high-grade squamous intraepithelial lesion (H-SIL/CIN2+) or adenocarcinoma. Two negative controls were included: one patient with post-menopausal bleeding and another from whom an inflammatory cervical sample was taken without conization. Prior to conization, a gynaecologist collected two cervical samples, fixed in PS or STM, from each patient. All but four cases were tested for panHPV (GP5+/GP6+) and specific hr-HPV subtypes (HPV16, 18, 31,33) by PCR using both media and all were processed for HC2. This study demonstrates that both HPV detection techniques work with PS, showing a specificity of 78.3% for HC2 and 92.8% for PCR compared to 83.8% for HC2 and 92% for PCR using STM. The efficacy of detecting HPV in PS-preserved H-SIL/CIN2+ was very high (96% for PCR using PS and 86% for HC2 using PS), which was in the same range as for PCR using STM, and which was only slightly lower than for HC2 using STM (96% and 89%, respectively). The differences were not statistically significant. It is concluded that ThermoFisher's PS is a valid liquid-based cytology medium for cervical samples, convenient for HPV testing by PCR with GP5+/GP6+ primers and by the HC2 assay.

Optimizing specimen collection and laboratory procedures reduces the non‐diagnostic rate for endoscopic ultrasound‐guided fine‐needle aspiration of solid lesions of the pancreas

“…16 The endoscopists emptied the syringe and rinsed the needle in a vial containing a fixative. A maximum volume of 4 ml of the vial content was used to prepare one slide by cytocentrifugation in the laboratory.…”

Immune cell infiltration in head and neck squamous cell carcinoma and patient outcome: a retrospective study

“…In p16-sg samples, the presence of HPV was investigated from about ten 7-lm-thick dewaxed, rehydrated and proteinase K-treated tumor sections using PCR after DNA extraction with QIAMP DNA mini Kit (Qiagen, Valencia, CA) according to manufacturer's instructions [see reference 21]. In addition to HPV types 16, 18, 31, and 33 [see reference 21], samples were also screened for HPV-6 and -11 (primers: sens 5 0 -TTGCTGTGGATGTGA CAGCA-3 0 ; anti-sens 5 0 -GGTGCGCaGATGGGACACAC-3 0 ).…”

Comparison of the accuracy of Hybrid Capture II and polymerase chain reaction in detecting clinically important cervical dysplasia: a systematic review and meta‐analysis

“…We then excluded 175 citations by applying the inclusion criteria to the titles and abstracts and retrieved 95 full‐text articles for further review. The review process yielded 28 articles that met all inclusion criteria . In addition, we added one manuscript from our own group (Luu et al, submitted), which is currently under review (Fig.…”

“…A total of 22,417 women were described, with four articles [38,41,48,54] contributing fewer than 100 participants and 10 articles [14,21,25,26,31,37,39,42,52] (Luu et al, submitted), more than 500 participants. The article by Schiffman et al [42] had the largest sample size, with more than 3400 participants.…”

“…Over half of the articles (k = 16) [14,21,25,31,35,36,39,43,[45][46][47][48][49][50][51]54] reported results from Europe, with the remainder from Asia-Pacific (k = 5) [26,44,52,56,57], North America (k = 6) (Luu et al, submitted) [37,38,42,53,55], and South America (k = 2) [40,41]. Just under half (k = 14) [25, 38-41, 44-46, 49-53, 57] reported studies were conducted in screening settings; 10 were in a follow-up/diagnostic setting [21,26,31,35,42,47,48,[54][55][56] and the remainder (k = 5) (Luu et al, submitted) [14,36,37,43] were in both screening and follow-up/diagnostic settings. Most (k = 23) used a crosssectional study design [21, 25, 26, 31, 35, 36, 38-41, 44-54, 56, 57]; four, a cohort study design…”