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CRISPR based systems are enhancing the sensitivity, accuracy, and efficiency of molecular diagnostics.
FREMONT, CA: CRISPR-based technologies are appropriate for detecting the nucleic acid sequences of distinct pathogenic strains with single-base precision in molecular diagnostics. This is a significant benefit over the produced primers used in PCR to amplify target sequences, which are susceptible to non-specific amplification and off-target effects. Because of the ability to cleave or "cut" nucleic acids, CRISPR-based technologies are well-known for their usage in gene editing and gene therapy applications. Researchers have just started harnessing bacteria-derived technology to produce the next generation of speedy, precise, and low-cost molecular diagnostic devices that can identify everything, including antibacterial resistance, viral breakout and cancer-causing mutations in moving tumor cells.
How CRISPR is helping in diagnostics:
Cas13-based diagnostics have the benefit of directly targeting RNA. An RNA virus RT-PCR test necessitates amplification of the RNA into DNA before PCR, resulting in non-specific amplification and false-negative diagnosis. To identify target RNA molecules, the SHERLOCK platform employs Cas13a and a fluorescent reporter. To identify target RNA molecules, the SHERLOCK platform uses Cas13a and a fluorescent reporter. SHERLOCK has been recently improved to discover viral genetic material straight from patient serum, urine, and saliva samples in under two hours without using any other apparatus. Similarly, the DETECTR technique targets DNA with Cas12a to identify human papillomavirus in patient samples and is connected to a fluorescent reporter.
Many bacterial sequences linked to antibacterial drug resistance can be targeted using the FLASH-NGS technology, which combines Cas9 and NGS technologies. FLASH-NGS has been used to identify MRSA infections in clinical settings. Researchers recently used Cas12a to create a highly sensitive test for Mycobacterium tuberculosis (Mtb). The CRISPR-MTB assay demonstrated higher sensitivity than Mtb culture while maintaining a 98 percent specificity. Patients may receive reliable findings sooner since CRISPR-MTB does not require culture, in addition to improved sensitivity, which minimizes the risk of false negatives.
Cancer and genetics
Detecting genetic changes like single nucleotide polymorphisms (SNPs) presently requires benchtop tools or genome sequencing, but the specificity of CRISPR-based diagnostics could change that. CRISPR-based assays' high specificity and sensitivity would be particularly beneficial for detecting cell-free DNA and circulating tumor cells, which are present in highly minute levels in serum and require a highly sensitive test.
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