Rapid point-of-care detection of BK virus in urine by an HFman probe-based loop-mediated isothermal amplification assay and a finger-driven microfluidic chip

View article
Microbiology

Main article text

 

Introduction

Materials & Methods

Primer design

Preparation of BKV standard

Reaction system of the HF-LAMP assay

Evaluation of the specificity and sensitivity of the LAMP assay

Nucleic acid extraction

Reaction with a commercial BKV qPCR kit (qPCR assay 1)

In house qPCR assay (qPCR assay 2)

Evaluation of BKV HF-LAMP using clinical samples

DNA extraction-free HF-LAMP

Portable finger-driven microfluidic chip

Results

Primer design for the HF-LAMP assay

Specificity and sensitivity of the BKV HF-LAMP assay

Evaluation of HF-LAMP using extracted DNA from clinical samples of HIV-1-infected patients

Clinical evaluation of the nucleic acid extraction-free BKV HF-LAMP assay using clinical samples from HIV-1 infected patients

Clinical evaluation of the nucleic acid extraction-free BKV HF-LAMP assay using clinical samples from kidney transplant patients

POCT detection using a portable finger-driven microfluidic chip

Discussion

Conclusions

Supplemental Information

Supplementary Figures and Tables

DOI: 10.7717/peerj.14943/supp-1

Raw data from HF-LAMP and qPCR data

DOI: 10.7717/peerj.14943/supp-2

Additional Information and Declarations

Competing Interests

The authors declare there are no competing interests.

Author Contributions

Yongjuan Zhao performed the experiments, analyzed the data, prepared figures and/or tables, authored or reviewed drafts of the article, and approved the final draft.

Yi Zeng performed the experiments, prepared figures and/or tables, and approved the final draft.

Renfei Lu analyzed the data, authored or reviewed drafts of the article, and approved the final draft.

Zhiying Wang performed the experiments, analyzed the data, prepared figures and/or tables, and approved the final draft.

Xiaoling Zhang performed the experiments, prepared figures and/or tables, and approved the final draft.

Nannan Wu analyzed the data, authored or reviewed drafts of the article, and approved the final draft.

Tongyu Zhu analyzed the data, authored or reviewed drafts of the article, and approved the final draft.

Yang Wang designed the experiments, supervised the study, analyzed the data, authored or reviewed drafts of the article, and approved the final draft.

Chiyu Zhang conceived and designed the experiments, supervised the study, analyzed the data, authored or reviewed drafts of the article, and approved the final draft.

Human Ethics

The following information was supplied relating to ethical approvals (i.e., approving body and any reference numbers):

The Shanghai Public Health Clinical Center granted Ethical approval to carry out the study within its facilities (2018Y032) and registered and recorded at the Chinese Clinical Trial Registry (ChiCTR1800017947).

Data Availability

The following information was supplied regarding data availability:

The raw data of LAMP and qPCR are available in the Supplemental Files.

Funding

The study was supported by the Shanghai Science & Technology Innovation Action Program (21Y11900500) and the National Natural Science Foundation of China (81873621). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.

6 Citations 1,703 Views 143 Downloads

MIT

Your institution may have Open Access funds available for qualifying authors. See if you qualify

Publish for free

Comment on Articles or Preprints and we'll waive your author fee
Learn more

Five new journals in Chemistry

Free to publish • Peer-reviewed • From PeerJ
Find out more