Detection of human papillomavirus type 16 L1 gene methylation in leftover DNA samples obtained from Cobas 4800 HPV test
Keywords:
Human papillomavirus 16, pyrosequencing, L1 gene methylationAbstract
Background: It has been known that persistent infections with the high-risk human papillomavirus (HR-HPVs) is the main risk factor for cervical cancer progression. However, a minor of HR-HPV infected women developed cancer. Therefore, it is necessary to look for additional test to particularly identify HR-HPV infected women who are at higher risk to progress to cervical cancer quickly.
Objective: We aimed to detect HPV16 L1 gene methylation levels in the leftover DNA samples of HPV16 positive samples with cytology diagnosed as atypical squamous cells of undetermined significance (ASCUS) obtained from Roche Cobas 4800 HPV assay.
Methods: Methylation analysis within HPV16 L1 gene at CpGs 5600, 5606, 5609 and 5615 was performed by using pyrosequencing assay in 79 HPV16 positive samples.
Results: Reliable HPV16 L1 methylation results were obtained from 29 of 79 (36.7%) HPV16 positive samples; 27.6% (8/29) samples showed methylation 10.0% at CpG5600, followed by CpG5609 that 20.7% (6/29) samples showed methylation
10.0%. There was only one sample (3.4%) showed methylation percentage 10.0% at CpG5606 and CpG5615. One sample showed high methylation (> 40.0%) at all four CpGs.
Conclusion: In all, 36.7% (29/79) of HPV16 positive samples) of DNA samples have adequate amount of DNA for further HPV16 L1 gene hypermethylation. HPV16 L1 gene hypermethylation was found at CpGs 5600 and 5609 in these samples. The study suggested that women with ASCUS cytology with high HPV16 L1 gene hypermethylation ( 10.0%) might be of concern and should be useful for clinician to manage HR-HPV infected women.
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