TỐI ƯU HÓA PHẢN ỨNG REAL-TIME PCR PHÁT HIỆN VI KHUẨN STREPTOCOCCUS PNEUMONIAE GÂY VIÊM PHỔI CỘNG ĐỒNG Ở NGƯỜI LỚN

Abstract

Background: Streptococcus pneumoniae is the major contagious cause of communityacquired pneumoniae in adults. In microbiological testing, the culture method is considered the "gold standard"; however, this method requires time-intensive for results. This study to optimize a real-time PCR reaction using specific primers and probes targeting the cpsA gene to detect Streptococcus pneumoniae, the causative agent of community-acquired pneumonia in adults. Objectives: To develop a real-time PCR process to detect Streptococcus pneumoniae causing community-acquired pneumoniae in adults. Materials and methods: The study was conducted using the Streptococcus pneumoniae ATCC 6305 strain. Proceed to design specific primers and probes, then evaluate the annealing temperature, primer and probe concentrations, specificity and sensitivity of the reaction. Results: Optimization of primer pair and probe selection for the cpsA gene for use in the detection of Streptococcus pneumoniae. The annealing temperature was optimized at 57.1°C, with a probe concentration of 0.25 μM and primer concentration of 0.3 μM. The study demonstrated 100% specificity, a sensitivity limit of 6.4 × 10² copies, an accuracy of 98.22%, and an amplification efficiency of 97.1%. Conclusions: The study successfully optimized a real-time PCR assay targeting the cpsA gene for the rapid and specific detection of Streptococcus pneumoniae in cases of community-acquired pneumonia in adults.