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. Method: 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. Result: 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. Conclusion: Indicating that nanobacteria infection is related with placental calcification.