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As one important member of the biological sample library , blood samples are rich in biological molecules, which can be used for disease diagnosis , prognosis of disease staging , and prognosis et al.compared with tissue samples, blood samples have some advantages , such as easier to access, sampling in a row and rich test items and so on.However, after blood samples were collected, which need to be processed, packed and frozen in ice for long-term preservation container.Otherwise RNA in the blood sample is unstable and easily degradation in normal temperature transportation and preservation condition , thus subsequently affecting the molecular diagnostic biomarkers ′detection, analysis, research and development.In this paper, This review summarizes the current clinical and research work of routine blood collection and preservation methods effecting on blood RNA degradation and solution for avoiding these degrading which can help testing changings of blood biomarker more accurately .
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Objective To establish the method of identifying MRSA with Taqman-fluorescence quantitative PCR basing on mecA/nuc/fem B three gene combined detecting.Methods Taking the coagulase positive MRSA,which isolated from the clinical samples and confirmed by VITEK 2 compact microbial analyzer,as the research obj ect,designed mecA/nuc/fem B specific PCR primers and Taqman fluorescent probe by bio-software PrimerPremier 5 and Designer Beacon 7,FAM,HEX and ROX markers were used to label the fluorescent probe at 5’,and the end of 3’was labeled with BHQ1,detected by fluo-rescence quantitative PCR instrment.Results ①1 g/dl gel electrophoresis results showed that the primer’s specificity of mec A/nuc/fem B were good,and molecular weight of the amplification band consistent with the expected molecular weight and no non-specific amplification band.②Three genes were obtained specific amplification in a single tube single channel and single tube multiple channel detection in PCR,and the three gene amplification effect in a single tube single tube single chan-nel and multichannel PCR similar.Conclusion Successfully established a method of multi channel Taqman-probe fluores-cence quantitative PCR identification of MRSA,mec A/nuc/fem B combined detection can effectively differentiate coagulase negative and positive MRSA,improve the accuracy of identification.
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Objective To understand the relationship between the positive rate of Epstain‐Barr virus (EBV) and human cyto‐megalovirus (HCMV) nucleic acid detection with the age in the patients with aplastic anemia ,acute lymphoblastic leukemia and my‐eloid leukemia .Methods The patients clinically diagnosed as aplastic anemia ,acute lymphoblastic leukemia ,and myeloid leukemia after bone marrow puncture in the Navy General Hospital from January 2012 to December 2013were collected as the research sub‐jects .the nucleic acid test kit from DAAN Gene Company was adopted for conducting the EBV and HCMV nucleic acid testing and analyzing the relationship between the positive rate of the EBV and HCMV infection with the patient′s age .Results There were different testing positive rate among 3 kinds of hematonosis .As a whole ,the positive rate of HCMV was lower than that of EBV (15 .8% vs .43 .7% ) .The EBV nucleic acid detection positive rate had statistically significant difference among different ages of pa‐tients with aplastic anemia and myeloid leukemia .(P< 0 .01) .Conclusion For children patients with aplastic anemia disease and myeloid leukemia ,more attention should be paid to monitoring of EBV and HCM V nucleic acid .
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OBJECTIVE To study the pathogenicity of the Vibrio fluvialis isolated from the coastal seawater.METHODS Virulence experiment group:22 Kunming mice were divided into four subgroups in random:V.fluvialis was injected into abdominal cavity in the test subgroup.And Staphylococcus aureus and Escherichia coli were injected into the positive control subgroups,separately and aseptic physiological saline was injected into the negative control group.Wound infection group:22 SPF mice were divided into four supgroups in random after their legs were injured:the experimental supgroup(soaked in artificial seawater with V.fluvialis);the positive control groups(with S.aureus and E.coli,separately);the negative control group(soaked in aseptic artificial seawater).The general condition,blood routine,blood culture,organ culture and wound secretion culture of the mice were observed.The pathological analysis of the mice was taken after sacrifice on the 3rd day.RESULTS In virulence experiment group,among all the 7 mice′s blood culture of V.fluvialis supgroup,5 mice were found V.fluvialis positive after 12 h injection,and 2 mice kept on positive until 24 h.In wound infection group,pathological examination showed there were a large number of neutrophils distributed over the striated muscle of the injured sites and cellulitis formed.CONCLUSIONS The V.fluvialis isolated from the sea water has pathogenicity,and can cause wound) infection and septicemia when the concentration reached 106 CFU/ml.