Improving Laboratory Efficiency by Automation of Preanalytic Processing of ThinPrep Specimens for Real-Time PCR High-Risk HPV Testing

Abstract

Cervical specimens collected in liquid-based cytology (LBC) media are the most common sample type used for high-risk human papillomavirus (HPV) testing. Since preanalytic steps such as vortexing and decapping vials, liquid transfer to a sample input tube with matching unique identifier, and recapping the original vials are required for processing LBC samples prior to running the Abbott RealTime High Risk HPV assay (Abbott, Wiesbaden, Germany), a full manual execution can be complicated, especially in high-throughput diagnostic contexts. Here, a custom-configured worktable setup for the Tecan Freedom EVO (Tecan, Männedorf, Switzerland) designed to automate and control preanalytic steps for ThinPrep (Hologic, Marlborough, MA) samples was used to evaluate the impact of automated versus manual preanalytics. Archival results for manual processing of 226 samples were compared with those obtained with the Tecan protocol, observing a very good overall concordance for final assay interpretation (95.6%). High overall agreement (100%) resulted also from retesting 99 samples by both the preanalytical protocols. High reproducibility was observed analyzing 23 randomly selected samples by automated preprocessing in triplicate. Hence, the new configuration of the Tecan platform translates the manual steps required to process ThinPrep specimens into automated operations, controls sample identification, and allows for saving hands-on time, while maintaining assay reproducibility and ensuring reliability of results, making it suitable for screening settings.

Improving laboratory workflow through automated pre‐processing of SurePath specimens for human papillomavirus testing with the Abbott RealTime assay

“…The automation of pre‐processing ThinPrep samples using the Tecan Freedom EVO 150 instrument has previously been described in a study in Italy, demonstrating reproducibility and improved reliability of test results following automated pre‐processing of cervical specimens collected in original ThinPrep LBC vials after cytological analysis. Our study represents the first report on the performance of the automated Tecan EVO system for processing SurePath LBC prior to testing on the Abbott m 2000 System, comparing its performance to manual aliquoting for both triage and HPV primary samples, using sample material that was not depleted from cells or liquid by a cytology processor.…”

“…The Freedom EVO 150 instrument (Tecan EVO) is used by laboratories to automate a diverse range of applications and has previously been described in a study for pre‐processing ThinPrep LBC samples conducted in Italy . On a limited number of specimens, Barbieri et al demonstrated that automated pre‐processing leads to higher reproducibility of signal measured with RT hrHPV.…”

“…The Freedom EVO 150 instrument (Tecan EVO) is used by laboratories to automate a diverse range of applications and has previously been described in a study for pre‐processing ThinPrep LBC samples conducted in Italy . On a limited number of specimens, Barbieri et al demonstrated that automated pre‐processing leads to higher reproducibility of signal measured with RT hrHPV. The Cytology Centre at the Manchester University Foundation Trust has been the first to use a custom‐configured worktable set up (Figure ) and corresponding script of the Tecan EVO for handling SurePath LBC samples complying with the instructions for pre‐processing SurePath of the RT hrHPV assay: (1) prior to sample processing, the instrument matches barcodes of original SurePath vials and empty secondary sample input tubes for the Abbott m 2000 system (Abbott Sample Transfer Tubes) placed on an m 2000 sample input rack on the worktable; (2) LBC vials are vortexed for 20 seconds and (3) de‐capped, followed by (4) transfer of 600 μL of sample from the original vial to the secondary tube with identical barcode and (5) recapping of the original LBC vial.…”

Prevalence of abnormal pap smears in the western region of Saudi Arabia from 2010 to 2022

“…LBC also allows for the molecular testing of HPV from the same vial, as long as approximately 2 mL of sample is sent for molecular biology first (to avoid contamination), and then routine LBC preparation is carried out. [ 13 , 14 ] For laboratories accredited by CAP, the cytopathology checklist provides benchmarking data for the acceptable reporting-percentile rate (RPR) for each category and ASC/SIL ratio for each preparation type. [ 9 ] Our data, percentages and ASC/SIL ratios, as shown in [ Table 1 ], were within the 5–95 th RPR; due to copyright, we cannot share the CAP’s RPR in our study.…”

Liquid-based cytology screening in Gynecologic oncology

“…The steps involved in LBC are easily automated, making the process more efficient and less dependent on manual skills. This automation extends to screening, with several platforms allowing automated primary screening of LBC samples 22 . This dramatically increases the throughput of samples, essential in large-scale screening programs.…”