Elsevier

Analytica Chimica Acta

Volume 1212, 15 June 2022, 339937
Analytica Chimica Acta

VIR-CRISPR: Visual in-one-tube ultrafast RT-PCR and CRISPR method for instant SARS-CoV-2 detection

https://doi.org/10.1016/j.aca.2022.339937Get rights and content

Highlights

  • 1.An instant, ultrasensitive and visual method was proposed for SARS-CoV-2 detection.

  • 2.Ultrafast RT-PCR accomplished in only a few minutes was reported for the first time.

  • 3.It integrated all components in-one-tube and realized sampling-to-result detection.

  • 4.It is highly specific and could detect SARS-CoV-2 at nearly single molecule level.

  • It showed 100% positive predictive agreement during clinical samples detection.

Abstract

Until now, corona virus disease 2019 (COVID-19) remained to be an enormous threat for global health. As one viral illness induced by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), versatile, rapid and sensitive method for SARS-CoV-2 detection in early stage is urgently needed. Here, we reported an ultrasensitive and visual in-one-tube detection method which could be accomplished within half an hour from sampling-to-result. By integrating all reactions in one tube, liquid handling steps were omitted and amplicon contamination could be totally avoided. Magnetic beads were employed to achieve the fast extraction of viral nucleic acid and increase the sensitivity. Using portable thermocycler and blue light, the fluorescent results could be directly observed by naked eyes. The proposed method is of higher specificity and sensitivity, nearly at single molecule level. More important, results demonstrated 100% positive detection rate for 40 clinical samples, which was consistent with standard RT-PCR. Thus, our method is considerably simple, rapid, sensitive and accurate, holding great promise for the instant detecting of viruses including SARS-CoV-2 and the next generation of molecular diagnosis.

Graphical abstract

We proposed an ultrasensitive, instant and visual detection method termed as VIR-CRISPR for point-of-care SARS-CoV-2 detection. It integrated ultrafast RT-PCR and CRISPR in-one-tube with high specificity and detection sensitivity at nearly single molecule level, holding great promise for rapid clinical diagnosis.

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Keywords

CRISPR
SARS-CoV-2
Viral
Integration
Visual
Molecular diagnosis

Cited by (0)

(1) Rui Wang, Ph.D., is a research scientist in Human Phenome Institute of Fudan University. Her research is focused on rapid nucleic acid amplification and detection, microfluidics and nanotechnology.

(2) Yongfang Li, Ph.D. candidate at the department of Chemistry in Fudan University. Her current research interests are focused on the luminescent materials and their applications in in-vitro diagnosis.

(3) Yanan Pang received his Doctor's degree from China Second Military Medical University in 2019. Now, he is working as assistant research fellow in Changhai Hospital, Shanghai, China. He is major in non-coding RNA and the early diagnosis of cancer.

(4) Shihua Luo, Ph.D., is a a doctor at Department of Traumatology, Ruijin Hospital, School of Medicine, Shanghai Jiaotong University. He is mainly focus on disease treatment.

(5) Chunyan Qian, MD, the director of the clinical laboratory, Linping Campus, The Second Affiliated Hospital of Zhejiang University School of Medicine, focusing on the diagnosis of infectious diseases.

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