多种呼吸道病原微生物快速筛查技术的建立

目的解决临床急性呼吸道感染病原微生物快速筛查诊断的难题,建立一种能同时快速检测多种病原体的有效方法。方法采用多重PCR与Luminex xMAP多重分析技术平台相结合,以荧光微球为检测载体,建立快速、多重检测DNA或RNA技术平台。结果经14种(18个亚型)常见呼吸道病原微生物标准株检测,在反应体系中多重PCR产物只与相应的病原菌检测微球杂交,无非特异性反应;通过138例临床标本检测验证,肺炎患者支气管肺泡灌洗液标本中,细菌检出率占总病原微生物的37.68%,其中结核分枝杆菌占18.84%;病毒检出率占总病原微生物的44.93%,支原体、衣原体占17.39%;患者支气管肺泡灌洗液和鼻咽拭子标本中流感病毒A型的检出率分别为18.75%和26.92%。结论多重检测技术平台对急性呼吸道感染疾病的病原微生物快速检测具有实用价值,尤其在解决流行病病原学分析方面具有重要意义。

Detection Technology of Multiple Respiratory Pathogens in Clinical Specimens

OBJECTIVE To find a method for rapid detection of multiple respiratory pathogens in clinical specimens.METHODS The specific tiny microspheres, multiplex PCR technology and Luminex xMAP(flexible Multi-Analyte Profiling) were used in combination for rapid detection of 14 respiratory pathogens(18 typing) by DNA or RNA.RESULTS The specificity of the diagnostic system had been validated by 15 species of pathogens(19 typing) from ATCC.There was not cross-reaction in each other.In 138 samples collected from clinical respiratory tract infections,bacteria were detected in 26(37.68%) of 69 pathogens from 112 bronchoalveolar lavage,including 13(18.84%) of Mycobacterium tuberculosis and 44.93% of viral pathogens,and 17.39% of Mycoplasma pneumoniae and Chlamydia pneumoniae.Influenza A virus detected in bronchoalveolar lavage and 26 samples of nasopharyngeal swabs were 21 of 112(18.75%) and 7 of 26(26.92%),respectively by Luminex xMAP.CONCLUSIONS Luminex xMAP Multi-Analyte Profiling is highly sensitive and accurate,high throughput and increases assay speed for detecting multiple respiratory pathogens in clinical specimens.It is a useful tool for epidemiology yet.