反向点杂交快速检测流感嗜血杆菌

目的　研究以分子杂交为基础的流感嗜血杆菌 (Hi)的快速检测方法。方法　将Hi探针固定在尼龙膜上 ;用标记有地高辛的细菌通用引物聚合酶链反应 (PCR)扩增不同细菌DNA ,产物分别与点有探针的膜条杂交。结果　 8种细菌均扩增出 4 6 8bp目的带 ,而人白细胞及轮状病毒DNA扩增均阴性。 6株Hi杂交阳性 ,7株其他临床常见菌杂交结果阴性。结论　反向点杂交法简便、快速、准确 ,是检测Hi的一种理想方法。

Quick identification of haemophilus influenza with reverse dot blot

Objective To study the way of quick identification of haemophilus influenzae with reverse dot blot.Methods Oligonucleotide probe which is specially targeted to 16SrDNA of haemophilus influenzae was designed, then fixed the probe to nylon membranes, and hybridized with the production of gain with the universal primers.Results The universal primers could hybridize the target sequence from common pathogenic bacteria by PCR, and oligonucleotide probe could hybridize with haemophilus influenza specially and could not hybridize with other bacterias. It proved that the probe was of highly speciality.Conclusion Reverse dot blot is a good method of quickly identification of haemophilus influenzae.