Field-friendly and ultra-fast detection platform without nucleic acid extraction for virus detection

Gaihua Cao; Yifan Xiong; Yue Qiu; Jun Yang; Yu Wang; Fuping Nie; Danqun Huo; Changjun Hou

doi:10.1016/j.aca.2023.341865

Field-friendly and ultra-fast detection platform without nucleic acid extraction for virus detection

Anal Chim Acta. 2023 Nov 1:1280:341865. doi: 10.1016/j.aca.2023.341865. Epub 2023 Oct 3.

Authors

Gaihua Cao¹, Yifan Xiong¹, Yue Qiu¹, Jun Yang², Yu Wang², Fuping Nie³, Danqun Huo⁴, Changjun Hou⁵

Affiliations

¹ Key Laboratory for Biorheological Science and Technology of Ministry of Education, Bioengineering College of Chongqing University, Chongqing, 400044, PR China; State Key Laboratory of Cattle Diseases Detection (Chongqing) of Customs, Diagnosis and Testing Laboratory of Lumpy Skin Disease, Chongqing Customs Technology Center, Chongqing, 400020, PR China.
² State Key Laboratory of Cattle Diseases Detection (Chongqing) of Customs, Diagnosis and Testing Laboratory of Lumpy Skin Disease, Chongqing Customs Technology Center, Chongqing, 400020, PR China.
³ State Key Laboratory of Cattle Diseases Detection (Chongqing) of Customs, Diagnosis and Testing Laboratory of Lumpy Skin Disease, Chongqing Customs Technology Center, Chongqing, 400020, PR China. Electronic address: nie1626@163.com.
⁴ Key Laboratory for Biorheological Science and Technology of Ministry of Education, Bioengineering College of Chongqing University, Chongqing, 400044, PR China. Electronic address: huodq@cqu.edu.cn.
⁵ Key Laboratory for Biorheological Science and Technology of Ministry of Education, Bioengineering College of Chongqing University, Chongqing, 400044, PR China; Chongqing Key Laboratory of Bio-perception & Intelligent Information Processing, School of Microelectronics and Communication Engineering, Chongqing University, Chongqing, 400044, PR China. Electronic address: houcj@cqu.edu.cn.

PMID: 37858555
DOI: 10.1016/j.aca.2023.341865

Abstract

The polymeric chain reaction (PCR) has come under fire for being time-consuming, requiring expensive equipments, and requiring the extraction and purification of nucleic acids. Here, an ultra-fast and sensitive detection platform without nucleic acid extraction solved the above problems. Firstly, the RoomTemp Sample Lysis Kit released the nucleic acid in 3 min and removed the inhibition to facilitate the amplification reaction. What's more, ultra-fast PCR (UF-PCR) can complete 40 cycles in just 15 min and 50 s. To improve the sensitivity and provide more convenient reading modes, CRISPR/Cas12a was mediated to detect Lumpy skin disease virus (LSDV). The platform output fluorescence and Lateral flow dipstick (LFD) signals. The actual detection limit was 2 × 10¹ copies·μL^-1. The portable platform realized visualization, excellent sensitivity and quick speed. In summary, the field-friendly testing platform had great potential in practical testing.

Keywords: CRISPR/Cas12a; Lumpy skin disease virus (LSDV); No nucleic acid extraction; Nucleic acid detection; Ultra-fast quantitative real-time-PCR (qPCR).

MeSH terms

Animals
Cattle
Nucleic Acid Amplification Techniques*
Nucleic Acids*
Sensitivity and Specificity

Substances

Nucleic Acids