鹦鹉热衣原体rrn操纵元序列双重PCR检测方法的建立

Development of Duplex PCR method for detection of rrn operon sequence from Chlamydia Psittaci

为快速检测鹦鹉热衣原体,基于鹦鹉热衣原体rrn操纵元序列(即16S-23S rRNA间隔区基因和23S rRNA基因序列)建立了双重PCR法,对100份临床标本进行检测,并与单一PCR法、免疫荧光法进行了比较.结果显示:双重PCR法特异性与敏感性均高于单一PCR法、免疫荧光法.双重PCR平均阳性检出率为83.4%～97.2%,敏感性为500 fg/μL.结果显示鹦鹉热衣原体rrn操纵元序列双重PCR法不仅能够检测衣原体和非衣原体感染,而且能够同时实现鹦鹉热衣原体检测和鉴定,从而克服了单一引物应用时的不足,为未来鹦鹉热衣原体病分子流行病学调查及临床早期诊断提供了有效的检测手段.

To set up a method for rapid detection of Chlamydia psittaci, a duplex PCR method was developed according to the sequences of the rrn operon sequence of Chlamydia psittaci, that is, 16S ribosomal DNA sequence and 16S- 23S ribosomal DNA spacer sequences. The results of detection from clinical samples （n = 100） by duplex PCR were compared with those by general PCR and immune fluorescence method. It was found that the specificity and sensitivity of duplex PCR were higher than that of both general PCR and immune fluorescence method. The average positive rate of duplex PCR was 83.4 - 97.2 percent and the lowest detection level was 500 fg/μL. Through a single procedure of duplex PCR, not only Chlamydia or non-Chlamydia could be distinguished, but Chlamydia psittaci could be identified as well. The duplex PCR is rapid, reliable and simple and provides an effective method for the eptdemiology investigation and early diagnosis of Chlamydia psittacl.