Language
English
Publication Date
5-1-2024
Journal
Analytical Biochemistry
DOI
10.1016/j.ab.2024.115480
PMID
38331373
PMCID
PMC11899860
PubMedCentral® Posted Date
5-1-2025
PubMedCentral® Full Text Version
Author MSS
Abstract
Isothermal nucleic acid amplification methods have many advantages for use at the point of care. However, there is a lack of multiplexed isothermal amplification tests to detect multiple targets in a single reaction, which would be valuable for many diseases, such as infection with high-risk human papillomavirus (hrHPV). In this study, we developed a multiplexed loop-mediated isothermal amplification (LAMP) reaction to detect the three most common hrHPV types that cause cervical cancer (HPV16, HPV18, and HPV45) and a cellular control for sample adequacy. First, we characterized the assay limit of detection (LOD) in a real-time reaction with fluorescence readout; after 30 min of amplification the LOD was 100, 10, and 10 copies/reaction of HPV16, HPV18, and HPV45, respectively, and 0.1 ng/reaction of human genomic DNA (gDNA). Next, we implemented the assay on lateral flow strips, and the LOD was maintained for HPV16 and HPV18, but increased to 100 copies/reaction for HPV45 and to 1 ng/reaction for gDNA. Lastly, we used the LAMP test to evaluate total nucleic acid extracted from 38 clinical samples; compared to qPCR, the LAMP test had 89% sensitivity and 95% specificity. When integrated with sample preparation, this multiplexed LAMP assay could be useful for point-of-care testing.
Keywords
Female, Humans, Uterine Cervical Neoplasms, Human Papillomavirus Viruses, Sensitivity and Specificity, Papillomavirus Infections, Nucleic Acid Amplification Techniques, Human papillomavirus 16, Papillomaviridae, DNA, Viral, Human papillomavirus 18, loop-mediated isothermal amplification, HPV, lateral flow assay, multiplex
Published Open-Access
yes
Recommended Citation
Barra, Maria; Chang, Megan; Salcedo, Mila P; et al., "Single-Tube Four-Target Lateral Flow Assay Detects Human Papillomavirus Types Associated With Majority of Cervical Cancers" (2024). Faculty, Staff and Students Publications. 6356.
https://digitalcommons.library.tmc.edu/baylor_docs/6356