肺炎衣原体的体外传代培养  

The propagation culture of chlamydia pneumoniae in vitro

作　　者：杨彤[1] 张玉玲[1] 吴乃宝[2] 

机构地区：[1]吉林大学第四医院心内科,吉林省长春市130011 [2]吉林大学第四医院核医学科,吉林省长春市130011

出　　处：《中国厂矿医学》2008年第2期146-147,共2页Chinese Medicine of Factory and Mine

摘　　要：目的探讨肺炎衣原体(Chlamydia pneumoniae,Cpn)在体外连续扩增的方法。方法通过Cpn感染HEp-2细胞,采用高低速反复离心纯化的方法,并经Giemsa染色及DNA聚合酶链式反应(polymerase chain reaction,PCR)检测肺炎衣原体的DNA。结果Cpn感染HEp-2细胞可见包涵体形成,Cpn连续传三代后,应用Cpn特异性基因片段进行检测扩增,结果均可见437bp特异性扩增阳性带。结论Cpn能有效地在体外连续扩增。Objective To study the method of continuous amplified culture of chlamydia pneumoniac（Cpn） in vitro. Methods Using Robins＇method after infected by Cpn（ CWL-029）, HEp-2 cell was cultivated in 25 cm^2 flask and repeatedly centrifuged and purified, then Giemsa staining and polymerase chain reaction（PCR） were performed. Primers were HL-1 （5′-GTTGTTCATGAAGGCCTACT-3′） and HR-1 （5′- TGCATAACCTACGGTGTGTT-3′）. A 437bp DNA fragment of Cpn-specific DNA sequence was arnplified. Results Inclusion body could be seen in HEp-2 cell after infected by Cpn. After going down to three generations, Cpn-specific gene fragment was detected and amplified. The resuhs demonstrated that specific 437 bp positive zone was seen. Condusions Cpn can continuously be propagated in vitro.

关 键 词：肺炎农原体 HEP-2细胞 聚合酶链式反应 GIEMSA染色 

分 类 号：R518[医药卫生—内科学] R56[医药卫生—临床医学]