Analytical evaluation of the automated genotyping system (GenPlex) compared to a traditional real‐time PCR assay for the detection of high‐risk human papillomaviruses

Abstract

The detection of high‐risk human papillomaviruses (HPVs) is crucial for early screening and preventing cervical cancer. However, the substantial workload in high‐level hospitals or the limited resources in primary‐level hospitals hinder widespread testing. To address this issue, we explored a sample‐to‐answer genotyping system and assessed its performance by comparing it with the traditional real‐time polymerase chain reaction (PCR) method conducted manually. Samples randomly selected from those undergoing routine real‐time PCR detection were re‐analyzed using the fully automatic GenPlex® system. This system identifies 24 types of HPV through a combination of ordinary PCR and microarray‐based reverse hybridization. Inconsistent results were confirmed by repeated testing with both methods, and the κ concordance test was employed to evaluate differences between the two methods. A total of 365 samples were randomly selected from 7259 women. According to real‐time PCR results, 76 were high‐risk HPV negative, and 289 were positive. The GenPlex® system achieved a κ value greater than 0.9 (ranging from 0.920 to 1.000, p < 0.0001) for 14 types of high‐risk HPV, except HPV 51 (κ = 0.697, p < 0.0001). However, the inconsistent results in high‐risk HPV 51 were revealed to be false positive in real‐time PCR by other method. When counting by samples without discriminating the high‐risk HPV type, the results of both methods were entirely consistent (κ = 1.000, p < 0.0001). Notably, the GenPlex® system identified more positive cases, with 73 having an HPV type not covered by real‐time PCR, and 20 potentially due to low DNA concentration undetectable by the latter. Compared with the routinely used real‐time PCR assay, the GenPlex® system demonstrated high consistency. Importantly, the system's advantages in automatic operation and a sealed lab‐on‐chip format respectively reduce manual work and prevent aerosol pollution. For widespread use of GenPlex® system, formal clinical validation following international criteria should be warranted.