CRISPR-based biosensing is prospective for rapid and sensitive diagnosis of pediatric tuberculosis

Abstract

Pediatric tuberculosis (TB) is an important part of global TB prevention and control. Diagnosis of childhood TB still remains challenging when using conventional tests, due to the non-specific clinical manifestations and paucibacillary nature of the specimens. Thus, a sensitive, rapid and low-cost diagnostic test is of great demand. Benefiting from specific and rapid Cas-protein-based catalytic activities, CRISPR-based biosensing platforms (CRISPR platforms) are showing superiority in detecting pathogen nucleic acid traces in clinical samples. Based on their excellent sensitivity, and time and cost saved in existing research, this study aimed to highlight the potential of CRISPR platforms as a tool for diagnosing pediatric TB, and advocate for studies to evaluate its performance in specimens collected from children, especially noninvasive specimens. These platforms are also promising in identifying drug resistance and genotyping. All of the above will help early diagnosis of pediatric TB, thus guide reasonable treatment, and be significant in achieving the World Health Organization End-TB strategy.

Development and Evaluation of Rapid and Accurate CRISPR/Cas13-Based RNA Diagnostics for Pneumocystis jirovecii Pneumonia

“…first established a Cas13a-based molecular detection platform, termed Specific High-Sensitivity Enzymatic Reporter UnLOCKing, to detect specific strains of the Zika and Dengue viruses, to distinguish pathogenic bacteria, to genotype human DNA, and to identify mutations in cell-free tumor DNA ( Srivastava et al., 2020 ). Later, CRISPR technology has been increasingly used in molecular diagnostics, including African swine fever virus ( Wang et al., 2020 ) and tuberculosis ( Lyu et al., 2020 ). However, it has not been used in the detection of P. jirovecii .…”

Exploring nano-enabled CRISPR-Cas-powered strategies for efficient diagnostics and treatment of infectious diseases

“…Using 16S rRNA CRISPR/Cas13-based biosensor, it is now possible to detect M. tb following reverse transcription is prior to and after amplification. In the meantime, preventing RNA depletion should be prioritized, which could restrict the CRISPR/Cas13 system's use in tuberculosis molecular diagnosis [163]. Hu et al (2020) outlined the prototype of DNA-AuNPs bioprobes using a freezing-based approach that was salt-free and did not require thiol manipulation.…”

Next-Generation Diagnostic with CRISPR/Cas: Beyond Nucleic Acid Detection

“…With dCas9, the first approach for TB detection was initiated following which the CRISPR/Cas12 effector was utilized as a pulmonary and extra-pulmonary TB-detection platform [ 9 , 59 , 68 , 110 , 111 ]. CRISPR-based detection eliminates pathogen cultures and provides faster results without requiring an expert technician.…”