聚合酶链反应检测流感嗜血杆菌

Study on detection of Haemophilus influenzae by polymerase chain reaction

以一对来自编码流感嗜血杆菌（Ｈｉ）外膜蛋白Ｐ６的基因序列的引物，用聚合酶链反应（ＰＣＲ）扩增Ｈｉ各血清型标准菌株及３０株临床分离菌株，均获阳性结果。而７种对照菌扩增结果均为阴性。用ＰＣＲ反应后琼脂糖凝胶电泳检测的方法，由蒸馏水稀释的菌液及鼻咽深部分泌物模拟标本中分别至少可以检测到３～５个及５０个Ｈｉ菌体。对４０例肺炎患儿鼻咽深部分泌物进行Ｈｉ检测，结果２０例ＰＣＲ阳性，其中１７例培养也阳性。本文作者还首次用ＰＣＲ法从６例肺炎患儿尿标本中检测到了Ｈｉ。研究表明，ＰＣＲ可用于Ｈｉ的检测，该方法敏感、特异、简便、快速，具有良好的应用前景。

By using a primer set selected from the sequence of a gene encoding for Haemophilus influenzae (Hi) outer membrane protein P6,polymerase chain reaction(PCR) were carried out to detect Hi.All the 6 reference strains of serotype a f and 30 field strains examined can be amplified.While all the other 7 genera of bacteria examined gave negative results.In the experiment,PCR DNA amplification was able to detect as few as 3 5 Hi cells from distilled water diluted bacterial liquid and 50 Hi cells from simulated nasalpharynx secretion sample by agarose gel electrophoresis.40 samples of deep nasalpharynx secretion collected from children with pneumonia were investigated by PCR and culture.20 were PCR positive for Hi and 17 of these were also positive in culture.In the study,we had also first detected Hi out of the urine samples of 6 children with pneumonia by PCR. The study shows that PCR is a sensitive and specific technique,and may be used as a rapid and early diagnostic tool for Hi.