Chinese Journal of Tissue Engineering Research ›› 2025, Vol. 29 ›› Issue (5): 1040-1049.doi: 10.12307/2025.279

Previous Articles     Next Articles

Evaluation of FTA-LAMP direct extraction method for extracting DNA from Streptococcus mutans 

Wang Yuehui1, Shang Jin1, Yang Chen1, Fu Dongge1, Cao Can1, Zhang Xiaodong1, Wang Jingfu1, 2   

  1. 1Department of Stomatology, General Hospital of Northern Theater Command, Shenyang 110016, Liaoning Province, China; 2Department of Maxillofacial Surgery, The Third Affiliated Hospital of Air Force Military Medical University, Xi’an 710004, Shaanxi Province, China
  • Received:2024-01-27 Accepted:2024-03-09 Online:2025-02-18 Published:2024-06-04
  • Contact: Zhang Xiaodong, MD, Chief physician, Department of Stomatology, General Hospital of Northern Theater Command, Shenyang 110016, Liaoning Province, China Co-corresponding author: Wang Jingfu, MD, Attending physician, Department of Stomatology, General Hospital of Northern Theater Command, Shenyang 110016, Liaoning Province, China; Department of Maxillofacial Surgery, The Third Affiliated Hospital of Air Force Military Medical University, Xi’an 710004, Shaanxi Province, China
  • About author:Wang Yuehui, Master’s candidate, Physician, Department of Stomatology, General Hospital of Northern Theater Command, Shenyang 110016, Liaoning Province, China
  • Supported by:
    Liaoning Provincial Science and Technology Plan (General Program), No. 2022-MS-043 (to CS)

PDF

133

Abstract: BACKGROUND: Streptococcus mutans is an important pathogen of dental caries, and timely detection of its levels is of great significance for early detection and treatment of dental caries. 
OBJECTIVE: To evaluate the effect of loop-mediated isothermal amplification (FTA-LAMP) direct extraction of Streptococcus mutans DNA. 
METHODS: (1) Bacterial suspensions containing ATCC standard strains (Streptococcus mutans) were prepared and inoculated into the brain-heart leachate medium. After mixed thoroughly, the mixture was then diluted in a 10-fold gradient into seven concentrations (4.2×107, 4.2×106, 4.2×105, 4.2×104, 4.2×103, 4.2×102, 4.2×10 CFU/ mL), two parallel controls were made for each dilution level, and sterile water was used as a blank control. (2) The DNA of Streptococcus mutans was extracted using FTA Elute card, boiling method, kit extraction and lysate extraction methods separately and then amplified using LAMP technology was amplified. A specificity test was also performed to compare the differences between the four DNA extraction methods.  
RESULTS AND CONCLUSION: The DNA extracted by all four methods met the requirements for LAMP amplification. Specificity test results showed that only Streptococcus mutans could specifically amplify the target gene. The detection limit value of the DNA concentration was 4.2×103 CFU/mL for the lysate method, 4.2×104 CFU/mL for the FTA Elute card extraction method, 4.2×106 CFU/mL for the kit extraction method, and 4.2×107 CFU/mL for the boiling method. In the other aspects of the four extraction methods, the kit extraction method had the highest experimental cost, number of steps and time; the other three methods had the same number of steps, with the FTA Elute card method requiring the least amount of instruments, the boiling method having the lowest single cost, and the lysate extraction method taking the least amount of time. Only a small amount of bacteria were needed for successful extraction using both the FTA Elute card and lysate extraction methods. Compared with the FTA Elute card method, the lysate extraction method was superior in terms of time, but it had a high single cost and required more equipment. To conclude, the FTA-LAMP technology established in this study has the advantages of ease of operation, high specificity, high sensitivity, and visualization, which is expected to be a new way for efficient extraction and detection of Streptococcus mutans.  

Key words: Streptococcus mutans, Loop-mediated isothermal amplification, FTA Elute card, DNA, visualization

CLC Number: