title: Isolation, cultivation and identification of nanobacteria from placental calcification.
abstract: OBJECTIVE
  The etiology of placental calcification (PC) is lack of research. To detect the bacterial infection mechanisms for PC, the experiment of isolating, culturing and identifying the nanobacteria in PC was done.
  
  
  METHODS
  The calcified placental tissues from 18 confirmed PC cases with normal placental tissue samples from 18 cases were studied by transmission electron microscopy (TEM), special nanobacterial culture methods, and identification of 16S rRNA sequence.
  
  
  RESULTS
  Under transmission electron microscope (TEM), Nanobacteria-like particles (NLP) in extra-cellular matrix (ECM) of calcified placental tissues were found, they were 50-500 nm in diameter, existed aggregation, among hydroxyapatite (HAP) crystals. Isolation and culture of NLP from the calcified tissues with methods described for nanobacteria were successful. All calcified placental tissue samples showed white granular deposition, which were firmly attached to the bottom of the culture tubes visible to the naked eyes. In the control group they could not be seen. According to 16S rRNA gene sequences analysis and was amplified adopting PCR and obtained 1407 bp fragment. Submit to GenBank after sequencing with accession number JN029830.
  
  
  CONCLUSIONS
  Indicating that nanobacteria infection is related with placental calcification.
authors.affiliation: Department of Pathogenic Biology, Chong Qing Medical University, Chongqing, People’s Republic of China. lhluhe@126.com
authors.forename: He
authors.initials: H
authors.lastname: Lu
authors.forename: Ya-nan
authors.initials: YN
authors.lastname: Guo
authors.forename: Sheng-nan
authors.initials: SN
authors.lastname: Liu
authors.forename: Hui
authors.initials: H
authors.lastname: Zhu
authors.forename: De-chun
authors.initials: DC
authors.lastname: Zhang
mesh_terms: D000328:Adult; D059587:Calcifying Nanoparticles / Q000032:analysis* / Q000235:genetics / Q000302:isolation & purification*; D002114:Calcinosis / Q000382:microbiology* / Q000473:pathology; D018929:Cell Culture Techniques; D002478:Cells, Cultured; D005260:Female; D006801:Humans; D008828:Microbiological Techniques; D046529:Microscopy, Electron, Transmission; D010802:Phylogeny; D010920:Placenta / Q000382:microbiology / Q000473:pathology / Q000648:ultrastructure; D010922:Placenta Diseases / Q000382:microbiology* / Q000473:pathology; D011247:Pregnancy; D012336:RNA, Ribosomal, 16S / Q000032:analysis; D055815:Young Adult
chemical_list: D059587:Calcifying Nanoparticles; D012336:RNA, Ribosomal, 16S