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- W2385874906 abstract "Objective:To establish a rapid method to detect drug-resistance genotypes of extended spectrum-βLactamases(ESBLs) produced by gram negative bacillus using the real-time fluorescence quantitative PCR.Methods:According to clinical common genotypes of ESBLs, SHV, TEM,CTX-M,OXA and their homology, 9 pairs of specific primers were designed including SHV, TEM, CTX-M-1, CTX-M-2, CTX-M-8, CTX-M-9, OXA-1, OXA-2 and OXA-10.To extract DNA template by boiling assay, and then establish and grade up SYBR GREEN I real-time fluorescence quantitative PCR reaction system, finally definite real-time fluorescence quantitative PCR method.Its precision and range of linearity were tested.With established assay 51 multi-drug resistant ESBLs-E.coli K.pneumoniae were detected and compared with improved three dimensional extract tests.Results:Except OXA-2, 8 genotypes SHV, TEM, CTX-M-1, CTX-M-2, CTX-M8, CTX-M-9, OXA-1 and OXA-10 were amplified by quantitative PCR from 39 ESBLs+ and 51 multi-drug resistant ESBLsE.coli K.pneumoniae and confirmed by sequence testing.The range of linearity was 3×103-3×108 copies/mL, r =-0.994 7.Repetitive experiments showed that the average coefficient of variation between-runs was 9.6% .Comparing with three dimensional extract test, there was no significant difference(χ2 = 1.125,P 0.05).Conclusion:Testing drug-resistance genotypes of ESBLs with SYBR GREEN I real-time fluorescence quantitative PCR is a rapid,specific and sensitive method, which is capable of inspecting genotypes of ESBLs from clinical strains." @default.
- W2385874906 created "2016-06-24" @default.
- W2385874906 creator A5014964945 @default.
- W2385874906 date "2009-01-01" @default.
- W2385874906 modified "2023-09-27" @default.
- W2385874906 title "Real-time Fluorescence Quantitative PCR for Detecting Extended Spectrum-β-Lactamases Genotype" @default.
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