A primer-probe composition and kit for synchronous detection of 8 fungi and parasite-related pathogens associated with digestive tract infections in children
A technology for simultaneous detection of gastrointestinal infections, applied in the field of detection kits, to achieve the effects of rapid detection, strong primer specificity, and wide application
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[0029] The kit consists of primer-probe mixture, RT-PCR buffer, mixed enzyme solution, positive quality control and RNase-free water; RT-PCR buffer includes PCR buffer, dATP, dUTP, dCTP, dGTP and MgCl 2 ; The mixed enzyme solution includes HotStart Taq enzyme, reverse transcriptase and UNG enzyme; the positive quality control contains plasmids or RAN particles corresponding to the amplified gene sequences of gastrointestinal pathogens.
[0030] The final concentration of the primer in the amplification system is 100-1000nM; the final concentration of the probe in the amplification system is 50-500nM.
[0031] The reaction system of the kit is 30 μL, specifically: 7 μL of nucleic acid template, 20 μL of RT-PCR buffer, 1 μL of mixed enzyme solution, and 2 μL of primer-probe mixture.
[0032] Kit operation and result determination:
[0033] (1) Add 7 μL each of the sample DNA / RNA co-extraction template (extracted from the stool of the patient), RNase-free water, and positive qua...
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