The invention discloses a reverse transcription primer which can be applied to relative quantification detection of relevant
cell factors of a Thl / Th2
cell, a plurality of associatively amplified
gene specific primers and a kit. The kit comprises a
reagent A: 50muL of 5muM reverse transcription primer, a
reagent B: 50muL of 20U / muL M-MuLV
reverse transcriptase, a
reagent C: 200muL of 5*reverse transcription
buffer solution containing 5mM of dNTP (deoxy-
ribonucleoside triphosphate) including dATP (
deoxyadenosine triphosphate), dCTP (
deoxycytidine triphosphate), dGTP (
deoxyguanosine triphosphate) and dTTP (deoxythymidine triphosphate)], 250mM of
Tris-HCl, 250mM of KCl, 20mM of MgCl2 and 50nM of DTT (
dithiothreitol), a reagent D: 2*PCR (
Polymerase Chain Reaction)
buffer solution, wherein 1mL of
buffer solution contains 100 U of taq
enzyme, 600nM of ROX (
roxithromycin), Green I dye, 2mM of dNTPs (including dATP, dCTP, dGTP and dTTP), 80mM of
Tris-HCl, 80mM of KCl, 40mM of (NH4)2SO4 and 6mM of MgSO4, and 24 primer-containing PCR 8
cascade reaction tubes. The entire set of kit provided by the invention can be applied to relative quantification of the
cell factors of the Thl / Th2 cell; moreover, relative quantification analysis can be performed on mRNA (
messenger ribonucleic acid) fragments of six types of cell factors of different types of T cells, and the
immune state of an individual can be estimated according to a result.