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A mismatch amplification mutation assay for specific detection of ciprofloxacin-resistant Neisseria meningitidis
Yusuke Ota1, Reina Okada1, Hideyuki Takahashi2
1Department of Molecular Microbiology, Graduate School of Medicine and Dental Science, Tokyo Medical and Dental University, Tokyo, Japan.
Abstract:
Meningococcal chemoprophylaxis for people in close contact with patients with invasive meningococcal disease (IMD) is necessary for preventing the spread of Neisseria meningitidis. Ciprofloxacin (CIP) is commonly used to treat IMD. However, CIP-resistant N. meningitidis isolates have rapidly evolved worldwide; therefore, rapid and accurate detection of CIP-resistant N. meningitidis is essential. We developed a mismatch amplification mutation assay for identifying gyrA substitutions T91I and D95Y, associated with reduced CIP susceptibility, using two primer sets to detect these variants. Comparison with gyrA sequencing data showed complete congruency. This method enables reliable detection of CIP-resistant N. meningitidis, thus leading to efficient management and control of IMD infections.
Insights
Rapidly detect ciprofloxacin-resistant Neisseria meningitidis strains. This new assay identifies key genetic mutations, aiding in controlling invasive meningococcal disease (IMD) spread and ensuring effective treatment.
Area of Science:
- Microbiology
- Molecular Biology
- Infectious Diseases
Background:
- Invasive meningococcal disease (IMD) requires chemoprophylaxis for close contacts to prevent Neisseria meningitidis spread.
- Ciprofloxacin (CIP) is a common treatment for IMD, but resistance is increasing globally.
- Accurate and rapid detection of CIP-resistant N. meningitidis is crucial for effective disease management.
Purpose of the Study:
- To develop a rapid diagnostic method for identifying ciprofloxacin resistance in Neisseria meningitidis.
- To detect specific gyrA gene mutations (T91I and D95Y) associated with reduced CIP susceptibility.
Main Methods:
- Development of a mismatch amplification mutation assay (MAMA).
- Utilized two primer sets to specifically target gyrA substitutions T91I and D95Y.
- Validated the assay by comparing results with gyrA sequencing.
Main Results:
- The developed MAMA demonstrated complete congruency with gyrA sequencing data.
- The assay reliably identifies Neisseria meningitidis isolates with gyrA mutations conferring reduced ciprofloxacin susceptibility.
- Successful detection of key genetic variants linked to CIP resistance.
Conclusions:
- The MAMA provides a reliable and efficient method for detecting CIP-resistant N. meningitidis.
- This assay facilitates timely and appropriate clinical decisions for IMD management.
- Improved diagnostics can lead to better control of invasive meningococcal disease outbreaks.
