Bacterial etiology for chronic villitis is not supported by polymerase chain reaction for 16S rRNA DNA
Bacterial etiology for chronic villitis is not supported by polymerase chain reaction for 16S rRNA DNA
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DOI：
10.1007/s10024-005-0412-1
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发表时间：
2005-12-01
影响因子：
1.9
通讯作者：
Howe, JG
中科院分区：
文献类型：
--
作者：
Ernst, LM;Crouch, J;Howe, JG
慢性绒毛炎的特征是绒毛被淋巴细胞、组织细胞和有时浆细胞浸润。在一小部分病例中，慢性绒毛炎区域内可发现感染原。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的直接传播，细菌微生物对培养特别有问题。为了验证这一假设，使用通用细菌16S rRNA DNA引物的聚合酶链反应（PCR）对从耶鲁病理学数据库中的胎盘中选择的慢性绒毛炎区域提取的DNA进行。首先通过检查苏木精和伊红染色的切片确认慢性绒毛炎的特定区域，然后从存档的石蜡块中取出。还制备了加标已知细菌计数的对照组织，以测试实验的灵敏度。将所有组织脱蜡、脱水并用蛋白酶K消化。使用Gentra Puregene试剂盒进行DNA提取。使用引物p11和p13对16S rRNA DNA进行PCR。扩增产物经琼脂糖凝胶电泳鉴定。对19例多灶性慢性绒毛炎的标本进行了研究。尽管样品和对照中有足够的DNA，但使用16S rRNA DNA的PCR引物，慢性绒毛炎标本均无可证实的产物。该检测试剂盒对每个样本约1500个细菌具有灵敏度。总之，这些数据不支持慢性绒毛炎的细菌病因。
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.