Hu Liuyang, Han Bing, Tong Qin, Xiao Hui, Cao Donglin
Department of Laboratory Medicine, Guangdong Second Provincial General Hospital, Guangzhou 510317, China.
The Second School of Clinical Medicine, Southern Medical University, Guangzhou 510515, China.
Can J Infect Dis Med Microbiol. 2020 Feb 26;2020:2697230. doi: 10.1155/2020/2697230. eCollection 2020.
. , , , , , , , and are primary respiratory bacterial pathogens contributing to morbidity and mortality in developing countries. This study evaluated the diagnostic performance of multiplex real-time PCR with fluorescence melting curve analysis (MCA) assay, which was used to detect eight respiratory bacterial pathogens simultaneously.
A total of 157 sputum specimens were examined by multiplex real-time with fluorescence MCA, and the results were compared with the conventional culture method.
Multiplex real-time PCR with fluorescence MCA specifically detected and differentiated eight respiratory bacterial pathogens by different melting curve peaks for each amplification product within 2 hours and exhibited high repeatability. The limit of detection ranged from 64 to 10 CFU/mL in the multiplex PCR system. Multiplex real-time PCR with fluorescence MCA showed a sensitivity greater than 80% and a 100% specificity for each pathogen. The kappa correlation of eight bacteria ranged from 0.89 to 1.00, and the coefficient of variation ranged from 0.05% to 0.80%.
Multiplex real-time PCR with fluorescence MCA assay is a sensitive, specific, high-throughput, and cost-effective method to detect multiple bacterial pathogens simultaneously.
[具体细菌名称]、[具体细菌名称]、[具体细菌名称]、[具体细菌名称]、[具体细菌名称]、[具体细菌名称]、[具体细菌名称]和[具体细菌名称]是发展中国家导致发病和死亡的主要呼吸道细菌病原体。本研究评估了采用荧光熔解曲线分析（MCA）的多重实时PCR检测方法的诊断性能，该方法用于同时检测8种呼吸道细菌病原体。
共157份痰标本采用荧光MCA多重实时PCR进行检测，并将结果与传统培养方法进行比较。
采用荧光MCA的多重实时PCR通过2小时内各扩增产物不同的熔解曲线峰特异性地检测和区分了8种呼吸道细菌病原体，且具有高重复性。多重PCR系统中的检测限为64至10 CFU/mL。采用荧光MCA的多重实时PCR对每种病原体的敏感性大于80%，特异性为100%。8种细菌的kappa相关性在0.89至1.00之间，变异系数在0.05%至0.80%之间。
采用荧光MCA的多重实时PCR检测方法是一种同时检测多种细菌病原体的灵敏、特异、高通量且经济高效的方法。
