Separation Of Pathogenic Bacteria And Research On The Sixteen Natural Plant Pathogen Microbes In The Cosmetics

With the rapid economic development and people’s living standard continues to improve, cosmetic safety issure are valued more and more people attention, microbiological testing of cosmetics is an important quality control project.Heiner pink, lavender, Osman are the Xinjiang Production of natural plant cosmetics market in recent years, sales of large, but the research on the kind of microorganisms in natural cosmetics little..In this study, 16 kinds of natural plant cosmetics as sample specimens, using membrane filtration method and inhibitors of pathogenic microorganisms were isolated samples tested by the VITEK 2 biochemical identification system for all strains isolated were identified and applied the United States Nccls promulgated by standard methods to identify pathogens have been doing drug susceptibility testing.Finally, isolated from Bacillus RpoB gene, Klebsiella pneumoniae pho E gene, Pseudomonas aeruginosa gyrA gene as a target gene, three pairs of specific primers were designed to establish the triple PCR reaction, causing rapid detection of these three molecular biology of bacteria.The test results shows that 16 kinds of samples through the use of filter method, preservatives interference suppression test, separated filter out typical bacteria, with satisfactory results.Using membrane filtration, flushing volume of 300 mL, seeded seven representatives of bacteria contamination recoveries were higher than 90%, so that membrane filtration method can effectively rule out bacteria preservatives interference.Screened for the detection of microorganisms were isolated, Gram staining, biochemical identification by VITEK 2 system identification to obtain a single purified strains: 5 E. faecalis, 1 Staphylococcus aureus, 3 Pseudomonas aeruginosa, 6 Klebsiella pneumoniae, 10 Bacillus licheniformis, 10 Enterobacter cloacae, 1 Streptococcus, 1 Enterococcus.6 kinds of Antibiotics and reagents susceptibility test results were significantly inhibitory diameter ring, and the strains showed varying degrees of resistance, no significant error determination result.The test established by triple PCR has a high sensitivity and specificity, artificially contaminated hair dye test sensitivity detection limit were Klebsiella pneumoniae 4.5×10 CFU / mL; Pseudomonas aeruginosa 6×102 CFU / mL; Bacillus licheniformis 1×10 CFU / mL. By annealing temperature test, optimize three pairs of primers, Mg2 +, Easy Taq DNA Polymerase and other parameters to determine the reaction system and response procedures triple the PCR. The results are as follows:10×Easy Taq Buffer(Mg2 +) 1.5 μL, 2.5 mM dNTPs 2.5 μL, on 10 μM gyrA each 1.0 μL downstream primer, 10 μMphoE upstream and downstream primers 1.0 μL, on 10 μM RpoB each 2 μL downstream primers 0.5μL, template DNA, 5 U / μL EasyTaq DNA Polymerase 0.5 μL, ddH2 O complement 25 μL; reaction program was: 94℃denaturation for 3 min; according to 94 45 s, 58 45 s, 72 ℃ ℃ ℃ 60 s were 30 cycles; last 72 for 7 min.℃...