scholarly journals Effectiveness of CRISPR based tools as alternative methods for diagnosing COVID-19

2021 ◽  
Vol 8 (3) ◽  
pp. 150-152
Author(s):  
Shefali Mehta ◽  
Aditi Mehta

The Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR)-Cas (CRISPR – Associated Proteins) systems are prokaryotic adaptive immune mechanisms that are used for cleaving the invading nucleic acids in nature. Due to this reason, the CRISPR-based tools are used in numerous applications like genome and transcriptome engineering, gene therapy, epigenome editing and many others. In addition, the issue of errors in these tests is also increasing at a very rapid pace, due to which their reliability and efficiency and effectiveness are getting severely hampered. Therefore, it may not be wrong to say that there is a still a lot more room for improvement and development to enable, widespread, rapid, and scalable testing of the COVID-19 virus. In this regard, the RT-qPCR tests can be used as a routine molecular diagnostic method for detecting any COVID-19 symptoms. However, there are various limitations that mar the use and implementation of the RT-qPCR tests. But a few of these shortcomings can be overcome by using alternative molecular diagnostic methods like the CRISPR based tests.

2016 ◽  
Vol 32 (5) ◽  
Author(s):  
Sang Hee Lee ◽  
Kwang Seung Park ◽  
Jung Hun Lee ◽  
Moonhee Park ◽  
Asad Mustafa Karim

2020 ◽  
pp. 1-4
Author(s):  
Kai-Chen Wang ◽  
Chiao-Yuan Fang ◽  
Chi-Chang Chang ◽  
Chien-Kuan Chiang ◽  
Yi-Wen Chen

2021 ◽  
Vol 32 ◽  
pp. S322
Author(s):  
Lui Ng ◽  
Ryan W.Y. Sin ◽  
Oswens S.H. Lo ◽  
Carlos K.H. Wong ◽  
Cindy L.K. Lam ◽  
...  

2021 ◽  
Author(s):  
Nathan Tanner ◽  
Yinhua Zhang

Reverse transcription loop-mediated isothermal amplification (RT-LAMP) has emerged as a viable molecular diagnostic method to expand the breadth and reach of nucleic acid testing, particularly for SARS-CoV-2 detection and surveillance. While rapidly growing in prominence, RT-LAMP remains a relatively new method compared to the standard RT-qPCR, and contribution to our body of knowledge on designing LAMP primer sets and assays can have significant impact on its utility and adoption. Here we evaluate 18 LAMP primer sets for SARS-CoV-2, comparing speed and sensitivity with different LAMP formulations and conditions across more than 5,000 RT-LAMP reactions and identifying several primer sets with similar high sensitivity for different SARS-CoV-2 gene targets. Significantly we observe a consistent sensitivity enhancement by combining primer sets for different targets, confirming and building on earlier work to create a simple, general approach to building better and more sensitive RT-LAMP assays.


2016 ◽  
Vol 54 (12) ◽  
pp. 2990-2999
Author(s):  
C. E. Scantlebury ◽  
G. L. Pinchbeck ◽  
P. Loughnane ◽  
N. Aklilu ◽  
T. Ashine ◽  
...  

Histoplasma capsulatumvar.farciminosum, the causative agent of epizootic lymphangitis (EZL), is endemic in parts of Africa. Diagnosis based on clinical signs and microscopy lacks specificity and is a barrier to further understanding this neglected disease. Here, a nested PCR method targeting the internal transcribed spacer (ITS) region of the rRNA operon was validated for application to equine clinical samples. Twenty-nine horses with signs of EZL from different climatic regions of Ethiopia were clinically examined. Blood samples and aspirates of pus from cutaneous nodules were taken, along with blood from a further 20 horses with no cutaneous EZL lesions. Among the 29 horses with suspected cases of EZL,H. capsulatumvar.farciminosumwas confirmed by extraction of DNA from pus and blood samples from 25 and 17 horses, respectively. Positive PCR results were also obtained with heat-inactivated pus (24 horses) and blood (23 horses) spotted onto Whatman FTA cards. Two positive results were obtained among blood samples from 20 horses that did not exhibit clinical signs of EZL. These are the first reports of the direct detection ofH. capsulatumvar.farciminosumin equine blood and at high frequency among horses exhibiting cutaneous lesions. The nested PCR outperformed conventional microscopic diagnosis, as characteristic yeast cells could be observed only in 14 pus samples. The presence ofH. capsulatumvar.farciminosumDNA was confirmed by sequencing the cloned PCR products, and while alignment of the ITS amplicons showed very little sequence variation, there was preliminary single nucleotide polymorphism-based evidence for the existence of two subgroups ofH. capsulatumvar.farciminosum. This molecular diagnostic method now permits investigation of the epidemiology of EZL.


Author(s):  
I. BALLESTEROS ◽  
M. P. MARTÍN ◽  
L. CERENIUS ◽  
K. SÖDERHÄLL ◽  
M. T. TELLERÍA ◽  
...  

2020 ◽  
Vol 22 (4) ◽  
pp. 368 ◽  
Author(s):  
Masashi Iijima ◽  
Kazuyoshi Shigehara ◽  
Hideki Igarashi ◽  
Koichi Kyono ◽  
Yasuo Suzuki ◽  
...  

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