Abstract
BackgroundTo develop a new 18 high-risk human papillomavirus (HR HPV) detection and genotyping assay, which is important to evaluate the risk degree of HR HPV for causing cancers.MethodsAll 18 HR HPV and β-globin relative DNA fragments were synthesized and cloned to a plasmid pUC57 to obtain their recombinant plasmids. Based on the matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) platform, each of the 18 HR HPV genotypes were investigated using their constructed recombinant plasmids. The new 18 HR HPV genotyping assay was tested using 356 clinical specimens and the results were compared to ones detected by the Roche Cobas 4800 HPV assay (Cobas). The discrepant results between two assays were resolved by sequencing and genotyping methods.ResultsThe new 18 HR HPV MALDI-TOF MS genotyping assay was developed using HPV recombination plasmids. The sensitivity was 103 to 102 copies/reaction for the all 18 HR HPV. This new developed HR HPV genotyping test was used to detect the clinical specimens. When the results on clinical samples detected by the new MALDI-TOF MS HPV test were compared with ones detected by the Roche Cobas 4800 HPV assay in terms of 14 HR HPV, the concordance was 80.1% (kappa coefficient, 0.60; 95% confidence interval [CI], 0.52–0.69). The discrepant results were resolved by sequencing and genotyping and suggests that the developed HR HPV assay is more sensitive and specific.ConclusionsThe new developed 18 HR HPV detection method based on MALDI-TOF MS platform is a high-throughput assay for the all 18 HR HPV genotypes and a powerful complement to current detection methods.
Highlights
To develop a new 18 high-risk human papillomavirus (HR Human papillomavirus (HPV)) detection and genotyping assay, which is important to evaluate the risk degree of High risk (HR) HPV for causing cancers
high-risk human papillomavirus (HR HPV) are associated with neoplastic lesions and carcinomas, while low risk (LR) HPV are mainly associated with benign lesions
Multiplex polymerase chain reaction (PCR) reactions were carried out using their HPV recombinant plasmids as templates based on the MALDI-TOF Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MS) platform
Summary
To develop a new 18 high-risk human papillomavirus (HR HPV) detection and genotyping assay, which is important to evaluate the risk degree of HR HPV for causing cancers. Persistent infection with high-risk types of human papillomavirus (HR HPV) has been established as a necessary, but insufficient, factor in the development of high-grade cervical dysplasia and cervical carcinoma [1]. HPV DNA could be detected in 99.7% of cervical carcinoma [2]. HPV DNA detection is of significance in the prevention and diagnosis of cervical cancer [4]. More than 100 HPV types have been identified and approximately 45 types can infect the genital tract [5]. HPV16 is the most prevalent HR HPV type everywhere in the world, HPV18 is slightly more prevalent in Europe and North America, HPV31 is more prevalent in South/ Central America, HPV33 and 45 are more prevalent in Africa, and HPV52 and 58 are more prevalent in Asia, such as China and South Korea [6, 7]
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