Author(s): Chen, FM (Chen, Fumin); Lyu, CA (Lyu, Chenang); Li, Z (Li, Zhao); Xiu, LS (Xiu, Leshan); Li, HM (Li, Huimin); Xie, Y (Xie, Yi); Cao, RZ (Cao, Runzhen); Hu, QQ (Hu, Qinqin); Yin, K (Yin, Kun)

Source: ADVANCED SCIENCE DOI: 10.1002/advs.202306612  Early Access Date: DEC 2023  

Abstract: Human norovirus (HuNoV) is the leading cause of nonbacterial acute gastroenteritis, which is highly infectious, rapidly evolving, and easily transmitted through feces. The accurate and early detection of HuNoV subtypes is essential for effective treatment, early surveillance, risk assessment, and disease prevention. In this study, a portable multiplex HuNoV detection platform that combines integrated microfluidics and cascade isothermal amplification, using a streamlined protocol for clinical fecal-based diagnosis is presented. To overcome the problems of carryover contamination and the incompatibility between recombinase polymerase amplification (RPA) and loop-mediated isothermal amplification (LAMP), a Dynamic confined-space-implemented One-pot RPA-LAMP colorimetric detection system (DORLA) is developed by creating a hydrogen bond network. The DORLA system exhibits excellent sensitivity, with detection limits of 10 copies mu L-1 and 1 copy mu L-1 for HuNoV GI and GII, respectively. In addition, a portable diagnostic platform consisting of a thermostatic control module and an integrated 3D-printed microfluidic chip for specific HuNoV capture, nucleic acid pretreatment, and DORLA detection, which enables simultaneous diagnosis of HuNoV GI and GII is developed. A DORLA-based microfluidic platform exhibits satisfactory performance with high sensitivity and portability, and has high potential for the rapid point-of-care detection of HuNoV in clinical fecal samples, particularly in resource-limited settings.

Herein, a dynamic confined space implemented one-pot recombinase polymerase amplification and Loop-mediated isothermal amplification colorimetric detection system (DORLA) is established by creating a hydrogen bond network, which can achieve a simultaneous diagnosis of Human norovirus GI and GII on the portable diagnostic platform.image

Accession Number: WOS:001128443300001

PubMed ID: 38126673

Author Identifiers:

Author Web of Science ResearcherID ORCID Number

Yin, Kun        0000-0001-6300-6985

eISSN: 2198-3844