Rapid diagnosis of seven high-risk human papillomavirus subtypes by a novel loop-mediated isothermal amplification...

Current human papillomavirus (HPV) detection methods require complex instruments, skilled staff and have a high cost. Therefore, novel testing approaches are needed which are easy to implement, highly sensitive, and low cost. Loop-mediated isothermal amplification (LAMP) is an isothermal amplification technique. In this study, according to the conditions in China, a novel LAMP method for detecting seven high-risk HPV subtypes (16, 18, 33, 39, 45, 52, and 58) was designed and evaluated. The DNA from plasmid and cervical specimens was extracted using Chelex 100 and measured by qPCR and LAMP assay. LAMP products were observed under ultraviolet light. HPV sequences were successfully amplified and a plateau time of 19-75 min was maintained. The concentration of positive reactions ranged between 20 copies/μL and 200000 copies/μL. Additionally, there was no cross-reactivity between HPV16, 18, 33, 39, 45, 52, 58, 31, 35, 45, 51, 56, 59, 66, or 68. For clinical samples, the LAMP assay had high sensitivity and specificity for HPV16, 18, 33, 39, 45, 52, and 58. However, 5% (72/1447) of the samples tested yielded false-positive results. In conclusion, the novel LAMP assay for HPV16, 18, 33, 39, 45, 52, and 58 has high sensitivity and specificity, a low cost, and is simple and rapid to perform. The LAMP assay can improve HPV detection in resource-limited settings, especially in primary care hospitals and rural areas.