Karunakaran Karuna P, Blanchard James F, Raudonikiene Ausra, Shen Caixia, Murdin Andrew D, Brunham Robert C
University of British Columbia Centre for Disease Control, Vancouver, Canada.
J Clin Microbiol. 2002 Nov;40(11):4010-4. doi: 10.1128/JCM.40.11.4010-4014.2002.
Recent whole-genome analysis has demonstrated limited genetic variation in Chlamydia pneumoniae, with one strain (AR39) containing a 4,524 nucleotide single-stranded DNA bacteriophage, PhiCpn1. Using PCR, reverse transcription (RT)-PCR, and Western blotting, we confirmed the presence and functional expression of PhiCpn1 in C. pneumoniae strain AR39 and its absence in strain CWL029. Six additional epidemiologically distinct clinical isolates of C. pneumoniae also did not contain PhiCpn1. We generated recombinant viral protein 1 (Vp1) from PhiCpn1 in Escherichia coli and showed that Vp1 antigen is highly immunogenic in mice and that murine antisera readily recognize native Vp1 from C. pneumoniae strain AR39 elementary bodies (EB). We developed an enzyme-linked immunosorbent assay (ELISA) to measure antibodies to recombinant Vp1 in human sera collected from 32 patients with abdominal aortic aneurysm (AAA) and 40 controls. Among the 72 subjects, 61 had C. pneumoniae EB antibodies shown by ELISA. Antibodies to Vp1 were found in 39 of the 61 (64%) seropositive individuals and were significantly correlated with AAA (adjusted odds ratio, 13.9; 95% confidence interval, 1.1 to 175). Our studies indicate that phage-containing strains of C. pneumoniae are uncommonly found by isolation but may commonly infect individuals with vascular disease.
最近的全基因组分析表明,肺炎衣原体的遗传变异有限,其中一个菌株(AR39)含有一种4524个核苷酸的单链DNA噬菌体PhiCpn1。我们通过聚合酶链反应(PCR)、逆转录(RT)-PCR和蛋白质免疫印迹法,证实了PhiCpn1在肺炎衣原体AR39菌株中的存在及其功能表达,而在CWL029菌株中不存在。另外6株在流行病学上不同的肺炎衣原体临床分离株也不含PhiCpn1。我们在大肠杆菌中表达了来自PhiCpn1的重组病毒蛋白1(Vp1),并表明Vp1抗原在小鼠中具有高度免疫原性,且鼠抗血清能轻易识别肺炎衣原体AR39菌株原体(EB)中的天然Vp1。我们开发了一种酶联免疫吸附测定(ELISA)法,以检测从32例腹主动脉瘤(AAA)患者和40例对照者采集的人血清中针对重组Vp1的抗体。在这72名受试者中,ELISA检测显示61人有肺炎衣原体EB抗体。在61名血清反应阳性个体中的39人(64%)发现了Vp1抗体,且与AAA显著相关(校正比值比为13.9;95%置信区间为1.1至175)。我们的研究表明,含噬菌体的肺炎衣原体菌株通过分离不常见,但可能普遍感染血管疾病患者。