Development of a Nucleic Acid Lateral Flow Immunoassay for the Detection of Human Polyomavirus BK.

Yi-Huei Huang,Wen-Zhi Lin,Juin-Hong Cherng,Shou-Ping Huang,Po-Da Hong,Kuan-Yi Yu,Yao-Wen Hung,Ming-Kung Yeh,Cheng-Che Liu,Hui-Wen Chuang

doi:10.3390/diagnostics10060403

Abstract

The BK virus (BKV) is an emerging pathogen in immunocompromised individuals and widespread in the human population. Polymerase chain reaction is a simple and highly sensitive method for detecting BKV, but it is time consuming and requires expensive instruments and expert judgment. The lateral flow assay, a rapid, low-cost, minimal-labor, and easy-to-use diagnostic method, was successfully applied for pathogen detection. In this study, we used oligonucleotide probes to develop a simple and rapid sandwich-type lateral flow immunoassay for detecting BKV DNA within 45 minutes. The detection limit for the synthetic single-stranded DNA was 5 nM. The specificity study showed no cross-reactivity with other polyomaviruses, such as JC virus and simian virus 40. For the Escherichia coli containing BKV plasmid cultured samples, the sensitivity was determined to be 107 copies/mL. The approach offers great potential for BKV detection of various target analytes in point-of-care settings.

Highlights

BK polyomavirus, known as the BK virus (BKV), is a small (40–45 nm in diameter), icosahedral and nonenveloped virus that contains an approximately 5-kbp circular double-stranded DNA
Depending on the elements of recognition used, Lateral flow assay (LFA) can be categorized as an antibody-based lateral flow immunoassay (LFIA), which uses a combination of antigens and antibodies, and nucleic acid lateral flow immunoassay (NALFIA), in which a single-stranded target analyte (DNA or RNA) forms a complex through complementary probe hybridization, generating detectable visual signals [23]
Data were collected from the National Center for Biotechnology Information (NCBI) database

Summary

Introduction

BK polyomavirus, known as the BK virus (BKV), is a small (40–45 nm in diameter), icosahedral and nonenveloped virus that contains an approximately 5-kbp circular double-stranded DNA. This virus is a member of the Polyomaviridae family and the Betapolyomavirus genus [1,2]. BKV remains as a latent infection in the epithelium of the renourinary tract [7,8]. BKV reactivates and replicates when individuals are immunosuppressed or immunocompromised due to cancer, pregnancy, diabetes, HIV infection, or transplants.

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