The invention discloses a primer
pool, a kit and a method for detecting
bloodstream infection by targeting high-
throughput sequencing based on a multiple amplification method. The primer
pool comprises a forward single-molecule tag primer and a reverse single-molecule tag primer, and each of the primers structurally comprises six parts, namely a forward single-molecule tag primer and a reverse single-molecule tag primer, and the forward single-molecule tag primer and the reverse single-molecule tag primer respectively comprises six parts: a tag universal amplification primer, a molecular tag UMI, a target specific primer sequence, an
RNA residue,
a DNA protection base and a blocking group sequentially arranged from a 5 '-3' end. The kit comprises the primer
pool and further comprises a unique
enzyme preparation and a reaction buffer. The kit can perform
absolute quantification, has high specificity, can significantly reduce primer dimers, can increase or decrease primers at any time according to expected target requirements, has strong expansibility, can simultaneously perform target co-detection of
RNA and
DNA in a
single tube, has comprehensive coverage, and reduces the detection cost.