CAJ | 학술논문

目的：建立TaqMan实时荧光定量PCR检测肺炎链球菌的方法，应用于临床儿童社区获得性肺炎（CAP）标本的快速筛查。方法根据 GenBank公布的肺炎链球菌自溶酶基因（lytA）设计引物和探针，建立实时荧光定量 PCR，并对体系进行优化；同时以痰培养法做双盲对照，应用于1504份 CAP患儿标本检测。结果 TaqMan实时荧光定量 PCR菌液的最低检出限可达18.75 cfu/PCR，无交叉反应，特异度好；对1504份儿童CAP标本检测，141份肺炎链球菌实时荧光定量PCR阳性，其中140份肺炎链球菌痰培养阳性，该方法灵敏度为100％，特异度为99.93％。从样品处理到结果报告仅需2.5 h。结论 TaqMan实时荧光定量PCR快速、简便、灵敏度高、特异度强，可用于儿童CAP中肺炎链球菌的初筛和指导抗生素的的合理使用。
목적：건립TaqMan실시형광정량PCR검측폐염련구균적방법，응용우림상인동사구획득성폐염（CAP）표본적쾌속사사。방법근거 GenBank공포적폐염련구균자용매기인（lytA）설계인물화탐침，건립실시형광정량 PCR，병대체계진행우화；동시이담배양법주쌍맹대조，응용우1504빈 CAP환인표본검측。결과 TaqMan실시형광정량 PCR균액적최저검출한가체18.75 cfu/PCR，무교차반응，특이도호；대1504빈인동CAP표본검측，141빈폐염련구균실시형광정량PCR양성，기중140빈폐염련구균담배양양성，해방법령민도위100％，특이도위99.93％。종양품처리도결과보고부수2.5 h。결론 TaqMan실시형광정량PCR쾌속、간편、령민도고、특이도강，가용우인동CAP중폐염련구균적초사화지도항생소적적합리사용。
Objective To develop TaqMan real-time PCR assay for detection and identification of streptococcus pneumonia iso-lated from children CAP.Methods Based on the sequences of lytA gene,primers and probe were designed and the assay was optimized.Then 1 504 sputum samples were detected by culture and the developed assay with double-blind testing.Results The lower limit of detection in the developed real-time PCR assay was 18.75 cfu/PCR,and had no cross reaction.141 strep-tococcus pneumonia strains were detected from 1 504 samples and 140 were isolated by culture.The whole process just nee-ded 2.5 h.Conclusion The established assay is rapid,simple,high sensitivity and specificity.It is not only valuable for the i-dentification of streptococcus pneumonia,but also provide evidence for antibiotic therapy.