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Rapid detection of the Streptococcus mutans cnm gene by loop-mediated isothermal amplification
Masae Kitagawa1, Kentaro Nagamine2, Hiroko Oka3
1Center of Oral Clinical Examination, Hiroshima University Hospital, Hiroshima, Japan.
Abstract:
This study investigated a method using loop-mediated isothermal amplification (LAMP) for the rapid detection of cnm-positive Streptococcus mutans (S. mutans) associated with cerebral microhemorrhage. LAMP amplified the cnm gene plasmid vector, but not human or microbial genomic DNA. The cnm DNA of the cnm-positive S. mutans strain was detected in saliva without DNA extraction after 1 day of culture. This method resulted in a cnm-positive rate of 26.4% in 102 samples, which was higher than that obtained with conventional PCR. In conclusion, LAMP may be used for the detection of cnm-positive S. mutans in a large number of samples.
Insights
Loop-mediated isothermal amplification (LAMP) offers rapid detection of cnm-positive Streptococcus mutans (S. mutans) linked to cerebral microhemorrhage. This method successfully identified S. mutans in saliva samples, outperforming traditional PCR.
Area of Science:
- Microbiology
- Molecular Biology
- Neurology
Background:
- Streptococcus mutans (S. mutans) is implicated in various infections.
- Cerebral microhemorrhage is a serious neurological condition.
- Rapid and accurate detection methods for S. mutans are crucial for timely diagnosis and treatment.
Purpose of the Study:
- To investigate the efficacy of loop-mediated isothermal amplification (LAMP) for detecting cnm-positive S. mutans.
- To assess the association of cnm-positive S. mutans with cerebral microhemorrhage.
- To compare LAMP with conventional PCR for S. mutans detection.
Main Methods:
- Loop-mediated isothermal amplification (LAMP) was employed to target the cnm gene of S. mutans.
- The method was tested on saliva samples from patients.
- DNA extraction was omitted for direct detection after a short culture period.
- Results were compared with conventional PCR.
Main Results:
- LAMP successfully amplified the cnm gene from a plasmid vector but not from human or microbial genomic DNA.
- cnm-positive S. mutans was detected in saliva samples after just one day of culture without DNA extraction.
- The cnm-positive rate using LAMP was 26.4% in 102 samples, exceeding the rate found with conventional PCR.
- LAMP demonstrated higher sensitivity and speed compared to conventional PCR.
Conclusions:
- LAMP is a viable and rapid method for detecting cnm-positive S. mutans.
- This technique shows promise for large-scale screening of S. mutans in clinical settings.
- LAMP facilitates the identification of S. mutans strains associated with cerebral microhemorrhage.
