Ernst Linda M, Crouch Jill, Rinder Henry, Howe John Greg
Department of Pathology, Room 5203, Children's Hospital of Philadelphia, 324 South 34th Street, Philadelphia, PA 19104, USA.
Pediatr Dev Pathol. 2005 Nov-Dec;8(6):647-53. doi: 10.1007/s10024-005-0412-1. Epub 2005 Jul 14.
Chronic villitis is characterized by chorionic villi infiltrated by lymphocytes, histiocytes, and sometimes plasma cells. In a small percentage of cases, an infectious agent can be demonstrated within areas of chronic villitis. However, the pathogenesis of most lesions is idiopathic. Chronic villitis may represent the direct spread of chronic endometrial infection by bacterial organisms that are particularly problematic for culture. To test this hypothesis, polymerase chain reaction (PCR) using primers for the universal bacterial 16S rRNA DNA was performed on DNA extracted from areas of chronic villitis selected from placentas in the Yale Pathology database. Specific areas of chronic villitis were first confirmed by examination of sections stained with hematoxylin and eosin and then removed from archived paraffin blocks. Control tissue spiked with known bacterial counts was also prepared to test the sensitivity of the experiment. All tissue was deparaffinized, dehydrated, and digested with proteinase K. DNA extraction was performed with the Gentra Puregene kit. PCR was done using primers p11 and p13 for the 16S rRNA DNA. The 233-bp amplified target product was identified by agarose gel electrophoresis. Nineteen specimens with multifocal chronic villitis without confinement to anchoring villi were studied. None of the chronic villitis specimens had a demonstrable product using the PCR primers for 16S rRNA DNA, despite adequate DNA in the samples and controls. The assay was sensitive down to approximately 1500 bacteria per specimen. In conclusion, these data do not support a bacterial etiology for chronic villitis.
慢性绒毛炎的特征是绒毛膜绒毛被淋巴细胞、组织细胞浸润,有时还有浆细胞浸润。在一小部分病例中,可在慢性绒毛炎区域内发现感染因子。然而,大多数病变的发病机制是特发性的。慢性绒毛炎可能代表慢性子宫内膜感染通过细菌病原体的直接传播,而这些细菌对于培养来说特别困难。为了验证这一假设,我们对从耶鲁病理学数据库中选取的胎盘慢性绒毛炎区域提取的DNA进行了聚合酶链反应(PCR),使用针对通用细菌16S rRNA DNA的引物。首先通过苏木精和伊红染色切片检查确认慢性绒毛炎的特定区域,然后从存档的石蜡块中取出。还制备了添加已知细菌计数的对照组织以测试实验的敏感性。所有组织均进行脱蜡、脱水,并用蛋白酶K消化。使用Gentra Puregene试剂盒进行DNA提取。PCR使用针对16S rRNA DNA的引物p11和p13进行。通过琼脂糖凝胶电泳鉴定233 bp的扩增靶产物。研究了19例多灶性慢性绒毛炎且不限于固定绒毛的标本。尽管样品和对照中有足够的DNA,但使用针对16S rRNA DNA的PCR引物,慢性绒毛炎标本中均未检测到可证明的产物。该检测方法对每个标本约1500个细菌敏感。总之,这些数据不支持慢性绒毛炎的细菌病因学。