Evaluation of a heat protocol with the Allplex HPV HR assay on the fully automated Seegene STARlet platform for detection of human papillomaviruses.

Abstract

Objective: Cervical cancer, primarily caused by persistent infections with high-risk human papillomaviruses (hrHPV), remains a significant global health issue. Effective screening methods are essential for early detection and prevention. This study evaluates a novel 'heat protocol' (HP) for pre-treatment of cervical samples in SurePath medium in the detection of hrHPV using the Seegene Allplex HPV HR Assay on the fully automated Seegene STARlet platform.

Methods: 50 cervical samples from women aged 21-69 were collected between March 2024 and June 2025 during routine screening at Sint-Trudo Hospital. Samples were pre-treated using either the HP (95 °C for 55 minutes on the Seegene STARlet) or the validated LB method. All were tested using the Allplex HPV HR Assay. The Alinity m assay served as reference. Precision, accuracy, and method comparison were assessed using clinical and external quality control samples. Statistical analyses included kappa agreement, non-inferiority testing, and Bland-Altman analysis of Ct values. .

Results: Our findings demonstrate that the HP shows higher sensitivity (96%) than both the LB pre-treatment (93%) and the external reference method (Alinity m). The HP also identified additional HPV genotypes not detected by the other methods, suggesting improved detection of low viral loads. Ct values were on average lower with HP, which supports this increased sensitivity. A non-inferiority analysis confirmed that HP is not inferior to LB.

Conclusion: The study confirms that the new HP offers excellent precision, accuracy, and sensitivity. While the LB pre-treatment remains a viable alternative, the HP's efficiency, reduced hands-on time and improved sensitivity make it the preferred method for our routine clinical practice.