两种不同基因引物检测痰液标本中肺炎链球菌的比较

目的建立cpsA和spn9802两种引物基因定量检测肺炎链球菌检测方法。方法根据基因库中两种基因的序列设计特异性引物,对127例肺炎患者痰液标本进行荧光定量PCR检测,比较两种方法检测的敏感性和特异性,并用传统痰培养方法同时检测标本。结果所有传统微生物检测肺炎链球菌阳性的12例标本在cpsA和spn9802检测中均为阳性;在传统微生物检测肺炎链球菌阴性的114例标本中cpsA法9例阳性,spn9802法29例阳性。统计分析表明,3种方法的阳性率比较存在显著性差异(P<0.05)。结论 cpsA和spn9802两种引物基因定量检测肺炎链球菌能提高肺炎患者痰液标本肺炎链球菌的阳性率。两种引物共同检测结果与培养结果相符,有一定的临床应用价值。

Comparison of Real-time PCR Methods by Two Different Primers for Identification of Stretococcus Pneumoniae

Objective To compare two real-time PCR methods by primer cpsA and spn9802 for detection and identification of stretococcus pneumoniae.Methods Two sets of primers targeting different genes of stretococcus pneumoniae were designed and synthesized.The sensitivity and specificity of real-time PCR were determined.127 sputum specimens from suspected pneumonia cases were detected by real-time PCR and bacterial culture simultaneously.Results Of the 127 sputum specimens,12 positive by culture were both positive by real-time PCR methods.Of the 115 negative by culture,12 were positive by cpsA real-time PCR and 29 were positive by spn9802 real-time PCR.The analysis showed that there was statistically significant difference between the three methods(P<0.05).Conclusion cpsA real-time PCR and spn9802 real-time PCR can effectively improve the rate of positivity for dignosis of suspicious pneumonia cases.Detecting sputum specimens with both cpsA and spn9802 can get the same results as bacterial culture,so it is useful for clinical diagnosis.