Rapid and Sensitive Detection of Human Astrovirus in Water Samples by Loop-mediated Isothermal Amplification with Hydroxynaphthol Blue Dye

Overview

Journal BMC Microbiol

Publisher Biomed Central

Specialty Microbiology

Date 2014 Feb 15

PMID 24524254

Citations 10

Authors

Bo-Yun Yang

Xiao-Lu Liu

Yu-Mei Wei

Jing-Qi Wang

Xiao-Qing He

Yi Jin

Zi-Jian Wang

Affiliations

Soon will be listed here.

Abstract

Background: The aim of this paper was to develop a reverse transcription loop-mediated isothermal amplification (RT-LAMP) method for rapid, sensitive and inexpensive detection of astrovirus.

Results: The detection limit of LAMP using in vitro RNA transcripts was 3.6 × 10 copies·μL⁻¹, which is as sensitive as the presently used PCR assays. However, the LAMP products could be identified as different colors with the naked eye following staining with hydroxynaphthol blue dye (HNB). No cross-reactivity with other gastroenteric viruses (rotavirus and norovirus) was observed, indicating the relatively high specificity of LAMP. The RT-LAMP method with HNB was used to effectively detect astrovirus in reclaimed water samples.

Conclusions: The LAMP technique described in this study is a cheap, sensitive, specific and rapid method for the detection of astrovirus. The RT-LAMP method can be simply applied for the specific detection of astrovirus and has the potential to be utilized in the field as a screening test.

Citing Articles

Recent Developments in Isothermal Amplification Methods for the Detection of Foodborne Viruses.

Suther C, Stoufer S, Zhou Y, Moore M Front Microbiol. 2022; 13:841875.

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Acute Infectious Gastroenteritis: The Causative Agents, Omics-Based Detection of Antigens and Novel Biomarkers.

Hasan H, Nasirudeen N, Ruzlan M, Mohd Jamil M, Ismail N, Abdul Wahab A Children (Basel). 2021; 8(12).

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A stoichiometric and pseudo kinetic model of loop mediated isothermal amplification.

Kaur N, Thota N, Toley B Comput Struct Biotechnol J. 2020; 18:2336-2346.

PMID: 32994892 PMC: 7493047. DOI: 10.1016/j.csbj.2020.08.020.

Next-generation amplicon sequencing identifies genetically diverse human astroviruses, including recombinant strains, in environmental waters.

Hata A, Kitajima M, Haramoto E, Lee S, Ihara M, Gerba C Sci Rep. 2018; 8(1):11837.

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Rapid and simple identification of carbapenemase genes, bla , bla , bla , bla and bla groups, in Gram-negative bacilli by in-house loop-mediated isothermal amplification with hydroxynaphthol blue dye.

Srisrattakarn A, Lulitanond A, Wilailuckana C, Charoensri N, Wonglakorn L, Saenjamla P World J Microbiol Biotechnol. 2017; 33(7):130.

PMID: 28585170 DOI: 10.1007/s11274-017-2295-5.

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