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Development and evaluation of a real-time multienzyme isothermal rapid amplification assay for rapid detection of

Yun Xing1, Zhixiong Duan2, Yuansu Jiang1

  • 1Department of Laboratory Medicine, Daping Hospital, Army Medical University, Chongqing, China.

Scientific Reports
|July 31, 2024
PubMed

Insights

A new rapid amplification assay accurately detects Streptococcus pneumoniae, a leading cause of childhood and elderly deaths. This simple method offers a promising tool for early diagnosis of pneumococcal infections, especially in resource-limited settings.

Area of Science:

  • Microbiology
  • Infectious Diseases
  • Molecular Diagnostics

Background:

  • Streptococcus pneumoniae causes severe infections like pneumonia, meningitis, and septicemia.
  • Despite vaccines, pneumococcal disease remains a major global health threat, particularly for young children and the elderly.
  • Early and accurate detection of S. pneumoniae is critical for effective patient management and improved outcomes.

Purpose of the Study:

  • To evaluate a novel real-time multienzyme isothermal rapid amplification (MIRA) assay for the detection of Streptococcus pneumoniae.
  • To assess the assay's performance in identifying S. pneumoniae and differentiating it from other bacterial species.
  • To determine the utility of the MIRA assay for rapid and reliable diagnosis in clinical settings.

Main Methods:

  • Design and optimization of a primer-probe set targeting the S. pneumoniae lytA gene.
  • Development and validation of a real-time multienzyme isothermal rapid amplification (MIRA) assay.
  • Testing the assay with 79 clinical isolates previously identified using the VITEK MS system.

Main Results:

  • The real-time MIRA assay demonstrated 100% sensitivity and 100% specificity in detecting Streptococcus pneumoniae.
  • The assay successfully identified S. pneumoniae from clinical isolates.
  • The optimized MIRA assay proved to be rapid, simple, and reliable.

Conclusions:

  • The developed real-time MIRA assay is a highly effective tool for the rapid and accurate detection of Streptococcus pneumoniae.
  • This assay shows significant potential for diagnosing pneumococcal infections, especially in resource-limited environments.
  • The MIRA assay can be applied for the detection of both S. pneumoniae and other non-S. pneumoniae bacterial species.

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