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
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页数:11
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