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A novel detection method based on MIRA-CRISPR/Cas13a-LFD targeting the repeated DNA sequence of Trichomonas vaginalis

Zhenke Yang1, Jinghui Wang1, Yiming Qi1

  • 1Xinxiang Key Laboratory of Pathogenic Biology, Department of Pathogenic Biology, School of Basic Medical Sciences, Xinxiang Medical University, Xinxiang, China.

Parasites & Vectors
|January 8, 2024
PubMed
Abstract

Insights

A new MIRA-CRISPR/Cas13a-LFD test accurately detects Trichomonas vaginalis, the leading cause of non-viral STIs. This rapid and convenient method offers improved diagnosis for vaginitis compared to existing techniques.

Area of Science:

  • Medical Diagnostics
  • Molecular Biology
  • Infectious Diseases

Background:

  • Trichomonas vaginalis is the most common non-viral sexually transmitted infection globally.
  • It is associated with pelvic inflammatory disease, adverse pregnancy outcomes, and increased HIV risk.
  • Current detection methods are expensive and complex.

Purpose of the Study:

  • To develop a novel, rapid, and accurate diagnostic tool for T. vaginalis detection.
  • To evaluate the performance of the new assay compared to existing methods.

Main Methods:

  • Development of a multi-enzyme isothermal rapid amplification-clustered regularly interspaced short palindromic repeats (MIRA-CRISPR)/Cas13a-lateral flow device (LFD) assay.
  • The assay targets a specific DNA sequence of T. vaginalis and operates at 37°C for approximately 1 hour.
  • Testing was performed on clinical samples and compared with microscopy, PCR, and culture.

Main Results:

  • The MIRA-CRISPR/Cas13a-LFD assay demonstrated a detection limit of 1 × 10⁻⁴ ng/μl.
  • The assay showed 100% sensitivity and excellent diagnostic agreement (kappa=1) with the gold standard culture method, similar to nested PCR.
  • No cross-reactivity was observed with DNA from other common microorganisms.

Conclusions:

  • The MIRA-CRISPR/Cas13a-LFD assay is a convenient, rapid, stable, and accurate diagnostic tool for T. vaginalis.
  • This method shows potential for improving vaginitis diagnosis and management.
  • It represents a significant advancement over current diagnostic techniques.
Keywords:
CRISPR/Cas13aDiagnosisLateral flow deviceTrichomonas vaginalisVisualized

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