Application of Hybridization Chain Reaction/CRISPR-Cas12a for the Detection of SARS-CoV-2 Infection

被引:4
作者
Sagoe, Kate Obaayaa [1 ]
Kyama, Mutinda Cleophas [2 ]
Maina, Naomi [3 ]
Kamita, Moses [4 ]
Njokah, Muturi [3 ]
Thiong'o, Kelvin [5 ]
Kanoi, Bernard N. [4 ]
Wandera, Ernest Apondi [4 ,6 ]
Ndegwa, Davies [7 ]
Kinyua, Dickson Mwenda [8 ,9 ]
Gitaka, Jesse [4 ]
机构
[1] Pan African Univ Inst Basic Sci Technol & Innovat, Dept Mol Biol & Biotechnol, POB 62000-00200, Nairobi, Kenya
[2] Jomo Kenyatta Univ Agr & Technol, Coll Hlth Sci, Dept Med Lab Sci, POB 62000-00200, Nairobi, Kenya
[3] Jomo Kenyatta Univ Agr & Technol, Coll Hlth Sci, Dept Biochem, POB 62000-00200, Nairobi 00200, Kenya
[4] Mt Kenya Univ, Directorate Res & Innovat, POB 342-01000, Thika, Kenya
[5] Kenya Govt Med Res Ctr, Ctr Biotechnol Res & Dev, POB 54840-00200, Nairobi, Kenya
[6] Kenya Govt Med Res Ctr, Ctr Virus Res, POB 54840-00200, Nairobi, Kenya
[7] Kenya Med Training Coll, Dept Med Lab Sci, POB 30195-00100, Nairobi, Kenya
[8] Meru Univ Sci & Technol, Dept Phys Sci, POB 972-60200, Meru, Kenya
[9] Kirinyaga Univ, Dept Pure & Appl Sci, POB 143-10300, Kerugoya 10300, Kenya
关键词
hybridization chain reaction (HCR); severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2); clustered regularly interspaced short palindromic repeats (CRISPR); COVID-19; point-of-care (POC); diagnostics; CRISPR-CAS12A; DESIGN; COST;
D O I
10.3390/diagnostics13091644
中图分类号
R5 [内科学];
学科分类号
1002 ; 100201 ;
摘要
Globally, the emergence of the coronavirus disease (COVID-19) has had a significant impact on life. The need for ongoing SARS-CoV-2 screening employing inexpensive and quick diagnostic approaches is undeniable, given the ongoing pandemic and variations in vaccine administration in resource-constrained regions. This study presents results as proof of concept to use hybridization chain reaction (HCR) and clustered regularly interspaced short palindromic repeats (CRISPR)/Cas12a complex for detecting SARS-CoV-2. HCR hairpin probes were designed using the NUPACK web-based program and further used to amplify the SARS-CoV-2 N gene in archived nasopharyngeal samples. The results were visualized using agarose gels and CRISPR Cas12a-based lateral flow strips. The assay was evaluated using the gold standard, real-time polymerase chain reaction (RT-PCR), as recommended by the World Health Organization (WHO). The results show the comparative efficiency of HCR to RT-PCR. This study shows that HCR and CRISPR are viable alternatives for diagnosing SARS-CoV-2 in samples.
引用
收藏
页数:11
相关论文
共 44 条
  • [21] Genomic characterisation and epidemiology of 2019 novel coronavirus: implications for virus origins and receptor binding
    Lu, Roujian
    Zhao, Xiang
    Li, Juan
    Niu, Peihua
    Yang, Bo
    Wu, Honglong
    Wang, Wenling
    Song, Hao
    Huang, Baoying
    Zhu, Na
    Bi, Yuhai
    Ma, Xuejun
    Zhan, Faxian
    Wang, Liang
    Hu, Tao
    Zhou, Hong
    Hu, Zhenhong
    Zhou, Weimin
    Zhao, Li
    Chen, Jing
    Meng, Yao
    Wang, Ji
    Lin, Yang
    Yuan, Jianying
    Xie, Zhihao
    Ma, Jinmin
    Liu, William J.
    Wang, Dayan
    Xu, Wenbo
    Holmes, Edward C.
    Gao, George F.
    Wu, Guizhen
    Chen, Weijun
    Shi, Weifeng
    Tan, Wenjie
    [J]. LANCET, 2020, 395 (10224) : 565 - 574
  • [22] Mathieu E, 2021, NAT HUM BEHAV, V5, P947, DOI 10.1038/s41562-021-01122-8
  • [23] Miti A, 2018, METHODS MOL BIOL, V1811, P115, DOI 10.1007/978-1-4939-8582-1_8
  • [24] Ngamsom B, 2022, medRxiv, DOI [10.1101/2022.01.06.22268835, 10.1101/2022.01.06.22268835, DOI 10.1101/2022.01.06.22268835]
  • [25] Low picomolar, instrument-free visual detection of mercury and silver ions using low-cost programmable nanoprobes
    Rana, Muhit
    Balcioglu, Mustafa
    Robertson, Neil M.
    Hizir, Mustafa Salih
    Yumak, Sumeyra
    Yigit, Mehmet V.
    [J]. CHEMICAL SCIENCE, 2017, 8 (02) : 1200 - 1208
  • [26] Advanced Molecular and Immunological Diagnostic Methods to Detect SARS-CoV-2 Infection
    Rotondo, John Charles
    Martini, Fernanda
    Maritati, Martina
    Caselli, Elisabetta
    Gallenga, Carla Enrica
    Guarino, Matteo
    De Giorgio, Roberto
    Mazziotta, Chiara
    Tramarin, Maria Letizia
    Badiale, Giada
    Tognon, Mauro
    Contini, Carlo
    [J]. MICROORGANISMS, 2022, 10 (06)
  • [27] Hybridization Chain Reaction Lateral Flow Assays for Amplified Instrument-Free At-Home SARS-CoV-2 Testing
    Schulte, Samuel J.
    Huang, Jining
    Pierce, Niles A.
    [J]. ACS INFECTIOUS DISEASES, 2023, : 450 - 458
  • [28] Hybridization chain reaction enables a unified approach to multiplexed, quantitative, high-resolution immunohistochemistry and in situ hybridization
    Schwarzkopf, Maayan
    Liu, Mike C.
    Schulte, Samuel J.
    Ives, Rachel
    Husain, Naeem
    Choi, Harry M. T.
    Pierce, Niles A.
    [J]. DEVELOPMENT, 2021, 148 (22):
  • [29] CRISPR Technology in Gene-Editing-Based Detection and Treatment of SARS-CoV-2
    Shademan, Behrouz
    Nourazarian, Alireza
    Hajazimian, Saba
    Isazadeh, Alireza
    Biray Avci, Cigir
    Oskouee, Mahin Ahangar
    [J]. FRONTIERS IN MOLECULAR BIOSCIENCES, 2022, 8
  • [30] Pathological Features and Neuroinflammatory Mechanisms of SARS-CoV-2 in the Brain and Potential Therapeutic Approaches
    Sodagar, Aisha
    Javed, Rasab
    Tahir, Hira
    Abd Razak, Saiful Izwan
    Shakir, Muhammad
    Naeem, Muhammad
    Yusof, Abdul Halim Abdul
    Sagadevan, Suresh
    Hazafa, Abu
    Uddin, Jalal
    Khan, Ajmal
    Al-Harrasi, Ahmed
    [J]. BIOMOLECULES, 2022, 12 (07)