A Versatile Nanowire Platform for Highly Efficient Isolation and Direct PCR-free Colorimetric Detection of Human Papillomavirus DNA from Unprocessed Urine

Abstract

Purpose: As human papillomavirus (HPV) is primarily responsible for the development of cervical cancer, significant efforts have been devoted to develop novel strategies for detecting and identifying HPV DNA in urine. The analysis of target DNA sequences in urine offers a potential alternative to conventional methods as a non-invasive clinical screening and diagnostic assessment tool for the detection of HPV. However, the lack of efficient approaches to isolate and directly detect HPV DNA in urine has restricted its potential clinical use. In this study, we demonstrated a novel approach of using polyethylenimine-conjugated magnetic polypyrrole nanowires (PEI-mPpy NWs) for the extraction, identification, and PCR-free colorimetric detection of high-risk strains of HPV DNA sequences, particularly HPV-16 and HPV-18, in urine specimens of cervical cancer patients. Materials and Methods: We fabricated and characterized polyethylenimine-conjugated magnetic nanowires (PEI/mPpy NWs). PEI/mPpy NWs-based HPV DNA isolation and detection strategy appears to be a cost-effective and practical technology with greater sensitivity and accuracy than other urine-based methods. Results: The analytical and clinical performance of PEI-mPpy NWs was evaluated and compared with those of cervical swabs, demonstrating a superior type-specific concordance rate of 100% between urine and cervical swabs, even when using a small volume of urine (300 µL). Conclusion: We envision that PEI-mPpy NWs provide substantive evidence for clinical diagnosis and management of HPV-associated disease with their excellent performance in the recovery and detection of HPV DNA from minimal amounts of urine samples.

One-Step Polymerase Chain Reaction-Free Nanowire-Based Plasma Cell-Free DNA Assay to Detect EML4-ALK Fusion and to Monitor Resistance in Lung Cancer

“…Taken together, these findings highlight the importance of identifying the variant to determine the best treatment strategy for ALK-positive NSCLC. [20] The NW-based cfDNA assay showed better feasibility for the detection of specific ALK fusion variants than genotyping methods used in other studies such as targeted next-generation sequencing, Sanger sequencing, and whole-exome sequencing.…”

“…Recently, we developed a new genotyping assay that accurately and rapidly detects small amounts of tumor-derived cell-free DNA (cfDNA) in various body fluids in patients with lung cancer or cervical cancer. [18][19][20] This assay isolates cfDNA in body fluids using nanowires (NW) and directly detects DNA variants or mutations without PCR amplification within 1 hour. Our previous studies demonstrated that this NW-based cfDNA assay can be used to detect low-abundance DNA variants in plasma cfDNA with high sensitivity and specificity.…”

“…In particular, due to the vast amount of raw data generated by NGS sequencing and its complexity, the role of bioinformatics experts is crucial for processing, interpreting, and analyzing complex bioinformatics data, ultimately obtaining biological specificity from sequenced samples. Recently, we developed a new genotyping assay that accurately and rapidly detects small amounts of tumor‐derived cell‐free DNA (cfDNA) in various body fluids in patients with lung cancer or cervical cancer [18–20]. This assay isolates cfDNA in body fluids using nanowires (NWs) and directly detects DNA variants or mutations without PCR amplification within 1 hour.…”

“…This assay isolates cfDNA in body fluids using nanowires (NWs) and directly detects DNA variants or mutations without PCR amplification within 1 hour. Our previous studies demonstrated that this NW‐based cfDNA assay can be used to detect low‐abundance DNA variants in plasma cfDNA with high sensitivity and specificity [18–20]. The NW‐based cfDNA analysis allows for a fast, cost‐effective, and real‐time multiplex analysis of target mutations, making it ideal for monitoring therapeutic responsiveness in a very simple manner.…”

Urine HPV in the Context of Genital and Cervical Cancer Screening—An Update of Current Literature

“…Two further recent studies have also been published by a research group that developed an ultrasensitive nanowire assay [ 38 , 39 ]. Both studies used polyethylenimine-conjugated nanowires (PEI-NWs) and implemented HPV DNA isolation and detection strategy in urine samples.…”