A kind of human sepsis pathogen detection kit and detection method
A detection kit and technology for sepsis, which can be used in biochemical equipment and methods, microbial determination/examination, resistance to vector-borne diseases, etc. Sensitivity and specificity issues, to achieve the effect of good sensitivity and specificity, high clinical application prospects, time and cost advantages
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Embodiment 1
[0022] Example 1: Composition of the kit
[0023] 1. Specific primer probes
[0024]
[0025]
[0026]
[0027] Note: where W is G or T: R is G or A; Y is C or T; M is A or C; S is G or C; D is A or G or T; K is G or T.
[0028] 2. The PCR reaction reagent is: TaqPath TM 1-Step Multiplex Master Mix (No ROX).
[0029] 3. Deionized water.
Embodiment 2
[0030] Example 2: DNA / RNA preparation
[0031] 1ml of human peripheral venous blood was collected according to routine medical operations, and EDTA was added for anticoagulation. Using Thermo's MagMAX TM CORE Nucleic Acid Purification Kit+MagMAX TM CORE Mastitis&PanbacteriaModule extraction kit to extract DNA and RNA.
Embodiment 3
[0032] Embodiment 3: PCR reaction system preparation
[0033] Prepare the PCR reaction system according to the following system:
[0034] A total of 7 reaction systems (the total reaction system is 20 μL), which are respectively S1-S7 in the above table, are composed of mixed internal reference primers and probes. PCR reaction reagent: 10 μL; the concentration of each primer and probe is 10 M, the volume of primers is 0.8 μL, and the volume of probe is 0.4 μL; sample template: 2 μL; deionized water makes up 20 μL.
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