聚合酶链反应检测肺炎衣原体方法的建立与初步应用

目的探讨聚合酶链反应（ＰＣＲ）技术在肺炎衣原体检测中的应用。方法参照Ｃａｍｐｂｅｌ报道的肺炎衣原体全基因序列、Ｇ＋Ｃ比例和次级结构设计一对引物，采用ＰＣＲ法检测肺炎衣原体ＤＮＡ及临床标本中的肺炎衣原体感染。结果经对肺炎衣原体、沙眼衣原体、大肠埃希菌、溶血性链球菌、白色念珠菌和结核分枝杆菌培养物进行ＰＣＲ扩增，结果只有肺炎衣原体出现单一４３７ｂｐ的扩增区带；敏感性试验可检测出１０ｆｇ的肺炎衣原体ＤＮＡ；１０份模拟标本均扩增出４３７ｂｐ区带；２２份小儿肺部感染的咽拭子标本中有５份为阳性，阳性率为２２．７％，而显微镜检查只有２例可见包涵体；１２份正常小儿咽拭子标本均为阴性。结论ＰＣＲ法检测肺炎衣原体，具有特异、敏感、快速、准确的特点，为肺炎衣原体感染的临床诊断提供了科学的依据

Detection of Chlamydiae pneumoniae with polymerase chain reaction

Objective To devolop a molecular biologic technique for detection of Chlamydiae pneumoniae (CP) with polymerase chain reaction. Methods A pair of primers were synthesized according to the designation of Campbell. DNA fragments of CP and digested from clinical samples were detected with PCR. Results 437 bp DNA fragments were amplified from CP, and they were not positive in chlamydia trachomalis, escherichia coli, mycobacterium tuberculosis, candida albicans, staphylococcus aureus and etc. The sensitivity could be improved to 10 fg. Its positive rate was 22.7% in the detection of nasopharyngeal secretion samples collected from 22 pneumonic infection infants, and it was nagtive in 12 normal infants. Conclusions This method is not only sensitive, rapid and specific, but also provides an etiological basis for diagnosis of CP pneumonia.