分子鉴别诊断(MDD)技术在呼吸道病原体检测中的应用

The application of molecular differential diagnoses platform in identification of the respiratory pathogens

目的:以靶序列富集多重PCR(Tem-PCR)和液相芯片(xMAP)检测技术结合,建立常见呼吸道病原体的分子鉴别诊断(MDD)平台,对其可靠性进行评估,并用于临床样品的检测。方法:采集苏州大学附属儿童医院呼吸科住院病例呼吸道灌洗液样品22例和苏州大学附属第一医院呼吸内科门诊病例咽拭子样品20例,使用Tem-PCR技术对样品的核酸进行多重扩增,以液相芯片技术进行高通量的多重检测。结果:对已知样品的检测表明该平台具有高度的特异性和敏感性。对42例临床样品进行检测的结果显示,22例呼吸道灌洗液的阳性检出率为63.6%。呼吸道灌洗液阳性检出率高于门诊病例咽拭子样品的阳性检出率,RNA病原体(以病毒为主)的阳性率高于DNA病原体(以细菌为主)的阳性率。结论:建立了呼吸道病原体的MDD平台,具备了对呼吸道传染性疾病的高通量快速诊断能力。

AIM： To estabilish a MDD（molecular differential diagnoses）platform for diagnosing the pathgens that may cause respiratory infection by combination of the advanced Tem-PCR（Target enriched multiplex PCR）with xMAP（multiple analyses profiling）,and to evaluate the reliability and further use the platform to test clinic samples.METHODS： 22 throat swab specimen from outpatient patients of respiratory department in First Affiliated Hospital of Suzhou University and 20 respiratory tract lavage fluid specimen from inpatients of respiratory department in Affiliated Children Hospital of Suzhou University were collected,and the nucleic acids of the samples were amplified by Tem-PCR and xMAP.RESULTS： Testing of the the known samples showed that the platform had excellent specificity and sensitivity.Testing of the clinic samples showed that the positive rate of the espiratory tract lavage fluid specimen was 63.6%,higher than that of the throat swab specimen,and that the positive rate of RNA pathgens was higher than that of DNA pathgens.CONCLUSION： A reliable MDD platform for detection of respiratory pathgens has been established with high-throughput detection capacity.