Articles Vol. 66 No. CĐ1-NCKH 24/01/2025

24. DETECTION OF METHICILLIN RESISTANT STAPHYLOCOCCUS AUREUS USING LOOP MEDIATED ISOTHERMAL AMPLIFICATION

Tran Hong Diem1,2, Nguyen Thi Truc Anh3,4, Nguyen Hong Phuc1, Vu Quang Hieu5,6
1 Viện Kỹ thuật công nghệ cao NTT, Trường Đại học Nguyễn Tất Thành
2 NTT Hitech Institute, Nguyen Tat Thanh University
3 University of Medicine and Pharmacy at Ho Chi Minh city
4 Trường Đại học Y Dược thành phố Hồ Chí Minh
5 Department of Biotechnology, NTT Hitech Institute, Nguyen Tat Thanh University
6 Khoa Công nghệ sinh học, Viện Kỹ thuật công cghệ cao NTT, Trường Đại học Nguyễn Tất Thành
Corresponding author: vqhieu@ntt.edu.vn
DOI: 10.52163/yhc.v66iCD1.1989
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Keywords
LAMP MRSA nasal swab isothermal amplification mecA gene
Abstract

Objective: To develop a loop-mediated isothermal amplification (LAMP) method for detecting Methicillin - resistant Staphylococcus aureus (MRSA) using the characteristic mecA gene, aimed at rapid MRSA detection in nasal swab samples.

Methods: An experimental study was conducted on 60 nasal swab samples from healthy individuals aged 18-25. Specific primers for the LAMP reaction were designed using primer design software. The LAMP reaction conditions were optimized in the laboratory to determine the temperature, specificity, and minimum DNA detection limit (LOD). The optimized LAMP reaction was then tested on 60 DNA extraction samples from nasal swabs, including 30 MRSA-positive and 30 MRSA-negative samples.

Results: Specific primers for mecA gene were successfully designed, and the amplification reaction was optimized at 65°C for 30 minutes. The LAMP method demonstrated high sensitivity and specificity, with the lowest detection limit (LOD) of 1 pg of DNA extracted from MRSA per reaction, and no cross-reactivity with other species was observed. When applied to 60 DNA samples extracted from nasal swabs (30 MRSA-positive and 30 MRSA-negative), the LAMP reaction achieved 100% sensitivity, 100% specificity, and a Kappa coefficient of 1, indicating a high degree of agreement compared to traditional microbiological culture and PCR methods.

Conclusions: The study successfully applied the LAMP method to detect the presence of MRSA in clinical samples with high sensitivity and specificity using the mecA gene. Furthermore, the study demonstrated a high level of agreement between the LAMP method, microbiological culture, and PCR diagnostics. LAMP is a promising method for MRSA detection, serving as a rapid diagnostic tool with high applicability to support effective treatment.

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