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- W2361411180 abstract "Objective To establish the rapid screening method for bacterial contamination of blood and related products.Methods Multiple sequences of 16S rDNAs from over 20 bacteria were aligned,and the universal primers were designed for fluorescence quantitative PCR(FQ-PCR)assay.Various genomic DNAs from 12 kinds of common pathogenic bacteria,2 kinds of fungi,1 kind of mycoplasma,1 kind of virus and human genomic DNA were used as templates of routine PCR to confirm the specificity of the universal primers.Genomic DNA of Staphylococcus aureus was extracted,and the target 16S rDNA fragment was amplified by PCR with universal primers to construct standard plasmid pMDT-Bfr.The standard quantitation curve was then established by using of the dilution gradients of pMDT-Bfr.To determine the sensitivity of FQ-PCR assay,dilution series of DNAs of Staphylococcus aureus and E.coli as representative strains for gram-positive and gram-negative bacteria,respectively,were first detected by FQ-PCR with SYBR Green I.To further verify the sensitivity of established FQ-PCR in the screening of bacterial contamination,DNAs extracted from blood samples spiked with series diluted Staphylococcus aureus and E.coli were used as templates.ResultsThe amplified products only have high homology with bacterial 16S rDNAs.The sensitivity of the FQ-PCR assay was 103 copies/μl of 16S rDNAs for both gram-positive and gram-negative bacteria.For the blood samples of artificial contamination,bacteria could be detected at concentrations of ≥105 CFU/ml.The above results suggest that FQ-PCR targeting the 16S rDNAs has high specificity and sensitivity.Conclusions The FQ-PCR assay targeting 16S rDNAs gene used to detect bacterial contamination of blood exhibits perfect application prospect in the future,because it is rapid,affordable,high sensitive and specific." @default.
- W2361411180 created "2016-06-24" @default.
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- W2361411180 date "2007-01-01" @default.
- W2361411180 modified "2023-09-23" @default.
- W2361411180 title "Rapid FQ-PCR screening for bacterial contamination of blood products based on 16S rDNAs" @default.
- W2361411180 hasPublicationYear "2007" @default.
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