Intelligent Screening Method in Oral Drugs by Visual Lamp System Specific Detection Technology

According to the relevant provisions of Chinese Pharmacepoeia
^[1]
, Staphylococcus aureus cannot be detected in oral drugs. In this experiment, the optimized LAMP reaction system was used to detect Staphylococcus aureus. Based on the thermostable nuclease gene (NUC) of Staphylococcus aureus. One external primer F3 and B3, and one internal primer FIP and BIP were designed in the conserved region based on the heat resistant nuclease gene (NUC) of Staphylococcus aureus. A visual loop-mediated isothermal amplification method for detection of Staphylococcus aureus was established by adding calcein Mn
²⁺
dye into the system; With this method, the specificity of 10 strains preserved in the laboratory was detected; And the sensitivity of the bacterial solution was detected by 10 times gradient dilution; Finally, the optimized LAMP reaction system was used to detect the commercial oral drugs, and the sensitivity of oral drugs contaminated by Staphylococcus aureus. The optimized lamp system was used to detect the specificity of 10 strains. The result was that the staphylococcus aureus, Pseudomonas aeruginosa and Streptococcus pneumoniae systems turned green, while the other systems were brown-yellow. At the same time, the results were verified by glue running Pseudomonas aeruginosa and Streptococcus pneumoniae showed trapezoid stripe in the UV Gel imaging system, and the rest had no stripe, with a specificity of 77%. The gradient diluted Staphylococcus aureus was detected based on lamp reaction, and the detection rate of the system was
$4.6 \times 10^{1}\ \text{CFU}/\text{mL}$
; The system was used for visual lamp detection of oral drugs sold in the market and not tested by the food and drug administration, and no Staphylococcus aureus was detected. However, visual LAMP detection of oral drugs artificially contaminated with S. aureus showed trapezoidal bands in experimental results, and the detection limit of s. aureus color in oral drugs detected by the system was up to
$5.3 \times 10^{2}\ \text{CFU}/\text{mL}$
. Conclusion: the visual lamp detection system of Staphylococcus aureus can be used to screen oral drugs containing Staphylococcus aureus.