Method for quantitatively detecting hantaan virus load through one-step fluorescence probe real-time quantitative reverse transcription polymerase chain reaction
A technology of reverse transcription polymerase and fluorescent probe, applied in the direction of microbial measurement/testing, microbial-based methods, biochemical equipment and methods, etc. Beach virus and other problems, to achieve the effect of high sensitivity, reduced operation steps, high sensitivity
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[0033] Example 1: Quantitative detection of Hantaan virus load in plasma of patients with hemorrhagic fever with renal syndrome:
[0034] Use the plasmid containing the 76-118S segment of the Hantaan virus standard strain to transcribe in vitro to obtain the S segment RNA of the 76-118 strain, quantify it accurately with an ultraviolet spectrophotometer, and calculate the molar concentration per unit volume based on its molecular weight as a standard; adjust the standard Concentration is 10 10 copies / ml, and 10 gradients were diluted 1:10 times as a standard curve.
[0035] After design and screening of primers and probes, the reverse transcription primers are random hexamers, in which:
[0036] The upstream primer is 5'-TACAGAGGGAAATCAATGCC-3';
[0037] The downstream primer is 5'-TGTTCAACTCATCTGGATCCTT-3';
[0038] The probe was 5'-(FAM)ATCCCTCACCTTCTGCCTGGCTATC(TAMRA)-3'.
[0039] Quantitative in vitro transcribed RNA was used as a standard, RNase-free water was used as...
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