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STUDY ON AMOTILE BACTERIA OF POSITIVE CULTURE IN NEWBORN: THE ANALYSIS OF PLASMID AND RESTRICTION ENZYME AND DETERMINATION OF OUTER MEMBRANE PROTEIN

ZHANG Wanming¹, WU Shixiao², LIU Guanxin²

1. Hubei Medical Staff College, Jingzhou, China

2. Children’s Hospital, Chongqing Medical University

Objective: To search for the reasons of high positive rate of amotile bacteria and the diagnosis of septicemia in newborn.

Methods: The blood was drawn from the different site of the newborn with septicemia and carried out blood culture. The drug sensitivity test had been done by the method of paper stripdiffusion. The plasmids of bacteria were extracted rapidly by modified Birnboim method and the plasmid analysis was carried out .The plasmids’s DNA of 35 epidemic strains was cut off by both restriction enzyme of Hind Ⅲ and EcoR1.The outer membrane protein (OMP) was determined by SDS-polyacrylamide gel electrophoresis.

Results: There are 51 patients with positive blood culture amotile bacterium, of them, pollution:35 cases(68.6%), septicemia: only 16 cases (31.4%),54.8%(57/104) strains bacteria have drug resistance to more of 12 drugs. 87.3%(165/189) strains bacteria have plasmids.They are cut off as 6 DNA fragments (1.9, 2.0, 4.0, 5.0, 8.5and 18kb) by Hind Ⅲ restriction enzyme .and as 5 DNA fragments (2.0, 2.6, 3.2, 6.3 and 22kb)by EcoRⅠ restriction enzyme, it is showed that they come from a same clone .The epidemic strain include 10 slips OMP, but non-epidemic strain have 11 slips OMP, increase a 25kd belt. The amotile bacteria with above mentioned plasmid spectrum, restriction enzyme spectrum and OMP spectrum are only seen in the air , therapeutic dish and syringe needle

Conclusion: The pollution is an important reason of amotile bacterium high positive rate in newborn. Diagnosing septicemia should depend on bacteria culture, plasmid analysis, restriction enzyme analysis of plasmid DNA, OMP determination and combining medical history and clinical manifestation.