posted on 2021-03-23, 05:44authored byJixue Weng, Nan Sheng, Runyuan Wang, Shuo Liang, Chen Wang, Xue Bai, Guohua Zhou, Bingjie Zou, Qinxin Song
Cervical
cancer is the fourth leading cause of death in women,
especially in developing countries. Specific and economic methodologies
for HPV typing are crucial in cancer diagnosis and further disease
control. However, routine assays based on real-time polymerase chain
reaction (qPCR) or DNA-chip hybridization are either incapable of
offering detailed subtype information or involve tedious open-tube
operations with the risk of cross-contamination from PCR amplicons.
Herein, we proposed a multiplex visualized closed-tube PCR (Multi-Vision)
for HPV typing. Using gold nanoparticle probes (AuNPs) as a color
change indicator combined with a Hamming distance 2 coding scheme,
13 high-risk HPVs and two subtypes associated with high-incidence
benign lesions were successfully typed by performing six closed-tube
PCRs. The assay demonstrates high specificity with no cross-reaction
among different subtypes under several artificial sample concentrations
(from 100 to 103 copies per reaction) and enables
highly sensitive detection of as low as 0.5 copies/μL. Further,
105 clinical samples were successfully analyzed using our method with
a high concordance rate of 99.05% (104/105) compared to a HPV typing
kit. The inconsistent sample was confirmed by sequencing to be consistent
with the typing results determined by our method, indicating that
Multi-Vision could be a useful tool for HPV detection, especially
in resource-limited regions.