CRISPR-Cas13a-based diagnostic method for Chlamydia trachomatis from nongonococcal urethritis

Bioanalysis. 2021 Jun;13(11):901-912. doi: 10.4155/bio-2021-0022. Epub 2021 May 7.

Abstract

Aim: Development of a routine screening technique for Chlamydia trachomatis infection. The proposed approach involves the CRISPR RNA (crRNA). In the presence of the target sequence, the RNase activity of the Cas13a protein is activated, and it cleaves RNA fluorescent probe so that fluorescence will be emitted. Results: The sensitivity of the detection based on CRISPR-Cas13a was 10 fM. The results obtained by CRISPR-Cas13a and quantitative polymerase chain reaction were closely correlated: χ2 = 81.798 (p < 0.001). Conclusion: The method can be carried out at room temperature and yields results within 2 h. The developed technique does not require expensive instruments and, thus, can meet the needs of community hospitals and other institutions for screening.

Keywords: CRISPR-Cas13a; Chlamydia trachomatis; nucleic acid detection; on-site detection.

MeSH terms

  • CRISPR-Cas Systems / genetics*
  • Chlamydia Infections / diagnosis*
  • Chlamydia trachomatis / isolation & purification*
  • Humans
  • Polymerase Chain Reaction
  • Urethritis / diagnosis*