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- W2350730596 abstract "Objective To establish dye real-time PCR method for the rapid detection of Oreintia tsutsugamushi(Ot).Methods A pair of specific primers were designed according to the 56kD outer membrane protein gene sequence of Oreintia tsutsugamushi in order to establish the dye method(SYBR green I) real-time quantitative PCR and its sensitivity and specificity were assessed.Various samples including standard Ot strains from chick embryo cultures,Ot isolates from animal organs and patients' blood were detected.Results A linear relationship between threshold cycle(Ct) of the quantitative real-time PCR and the DNA copy number could be demonstrated(r2= 0.99).Sensitivity assessment found that only 28 copies of the gene can be detected in each 20 μl PCR reaction tube,and the minimum detectable concentration was 2 copies / μl,presenting an increase by 2 orders of magnitude than the ordinary PCR and having good repeatability; The SYBR I dye method has good specificity for the other five kinds of Rickettsia and other pathogenic DNA templates presenting no specific amplification; 57 of the total 59 samples including the standard Ot strains from chick embryo cultures and Ot isolates from animal organs and patients' blood were detected positive.This method was also successfully employed to compare the quantity of Orientia tsutsugamushi in different organs such as blood,liver,spleen and kidney of the infected mice resulting that the most in spleen,then in liver and kidney,and the least in blood.Conclusion These results suggest that the dye quantitative real-time PCR assay presents highly specific and sensitive and is suitable for the detection of Orientia tsutsugamushi in various samples,so it may be employed successfully in the rapid diagnosis scrub typhus." @default.
- W2350730596 created "2016-06-24" @default.
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- W2350730596 date "2014-01-01" @default.
- W2350730596 modified "2023-09-28" @default.
- W2350730596 title "Detection of Orientia tsutsugamushi by dye quantitative real-time PCR assay" @default.
- W2350730596 hasPublicationYear "2014" @default.
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