Application of CRISPR detection technology in screening of stroke susceptibility genes

Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR) detection technique was used to screen stroke susceptibility gene methyltetrahydrofolate reductase (MTHFR), Shandong Province, to evaluate its predictive effect and clinical application value for stroke.

Sanger sequencing of MTHFR-C677T (rs 1,801,133) and A1298C (rs 1,801,131) was performed on blood samples of 635 patients with high-risk stroke, and the mutation of A1298C was detected by CRISPR. Based on the two sequencing results and the incidence of stroke, the predictive effect of MTHFR gene detection and the clinical application value of CRISPR technology were verified.

Sanger sequencing revealed a statistically significant difference (p < 0.05) in MTHFR C677T mutation frequency between the stroke group and controls, and between the high-risk group and controls, indicating an association of C677T polymorphism with stroke risk. Comparison between CRISPR and Sanger results showed similar mutation detection rates across groups (26.9% vs. 26.5% in stroke/TIA/high-risk groups; both 32.6% in controls). CRISPR demonstrated 97.6% sensitivity, 98.5% specificity, 98.3% concordance, and a Kappa value of 0.956. Furthermore, homocysteine level analysis indicated significant differences (p < 0.05) in MTHFR C677T and A1298C polymorphisms between individuals with normal and high homocysteine (HHcy) in stroke‑related groups.

CRISPR detection technique has high accuracy and is suitable for clinical application. The increase of homocysteine level may be one of the risk factors of stroke.