The SHERLOCK Platform: An Insight into Advances in Viral Disease Diagnosis

Abstract

Persistence and prevalence of microbial diseases (pandemics, epidemics) is the most alarming threats to the human resulting in huge health and economic losses. Rapid detection and understanding of the disease dynamics by molecular biotechnology tools allow for robust reporting, treatment and control of diseases. As per WHO, the optimal diagnostic approach should be quick, specific, sensitive, without a stringed instrument, and low cost. The drawbacks of traditional detection techniques promote the use of CRISPR-mediated nucleic acid detection methods such as SHERLOCK as detection method. It takes advantage of the unexpected in vitro features of CRISPR-Cas system to develop field-deployable sensitive detection tools. Previously, CRISPR-mediated diagnostic methods have extensively been reviewed particularly for SARS-COV-2 detection, but it fails to provide the insight into advances of this technique. This study is the first attempt to review the advances of SHERLOCK approach as diagnostic tool for viral diseases detection. Variations of SHERLOCK mechanism for improved efficiency are discussed. Particularly integrated SHERLOCK approaches in terms of extraction-free assay and Bluetooth-enabled detection are reviewed to access their feasibility for the development of simpler and cost-effective diagnostic toolkits. Insight in to perks and limitations of diagnostic methods indicates its potential as ultimate diagnostic instrument for disease management. Graphical Abstract

Recombinase polymerase amplification: method’s characteristics and applications in diagnostics of infectious diseases

“…Одним из наиболее активно развивающихся методов амплификации НК является «Specifc High-Sensitivity Enzymatic Reporter На основе системы CRISPR-Cas разработаны различные диагностические платформы для многих инфекционных патогенов, предложены разнообразные системы пробоподготовки: от применения термического лизиса в качестве экстракции НК до обработки магнитными частицами с целью накопления искомого материала и повышения чувствительности теста [25][26][27]. По данным публикаций, метод SHERLOCK обладает более высокой чувствительностью по сравнению с аналогичными подходами, предел обнаружения сочетанного использования технологий РПА и CRISPR-Cas составляет несколько копий мишени в реакции [24,28].…”

CRISPR Technology for Diagnosis: A Brief Review

“…Cas12 enzyme is currently used to diagnose Covid-19, HPV, cervix, and colorectal tumors. Other smaller enzymes, including Cas14 and CasX, are recently considered for utilizing in the SHERLOCK system as a detecting approach 6 .…”

Advancements in the synergy of isothermal amplification and CRISPR-cas technologies for pathogen detection

“…This platform is capable of detecting both RNA and DNA and boasts the advantages of versatility, convenience, and high sensitivity ( Kellner et al, 2019 ). The SHERLOCK detection process comprises three major steps: sample acquisition, amplification, and CRISPR-mediated collateral activity and detection ( Zahra et al, 2023 ). This technology is proving to be invaluable for rapid detection applications, playing a pivotal role in human health, particularly in distinguishing between different strains of viruses and other pathogenic variants ( Khan et al, 2021 ).…”

“…However, it is important to note that these technologies still face certain challenges, including multistep nucleic acid amplification and commercialization limitations ( Zahra et al, 2023 ). Nevertheless, the combined application of RPA/RAA technology and CRISPR-Cas has significantly altered the landscape by addressing environmental and site constraints, reducing costs, and enhancing sensitivity.…”