The invention belongs to the technical field of
virus in-vitro
nucleic acid detection and particularly relates to a digital PCR quantitative detecting kit for HBV (
hepatitis B
virus)
cccDNA and an application of the digital PCR quantitative detecting kit for HBV
cccDNA. A digital PCR quantitative detecting probe composition of the HBV
cccDNA is firstly designed, and the kit comprises a container for detecting the probe composition and a container for a purifying
reagent containing liver puncture tissue
DNA. Using of the kit comprise the following steps of extracting the
DNA in the liver puncture tissue, and adopting PSAD (
prostate specific
antigen density)
enzyme to digest the HBV cccDNA; by taking a digital PCR detecting
chip as a reaction container and adopting a complex fluorescent multiple PCR primer, amplifying target
DNA in detecting micro pores of the digital
PCR chip; and obtaining the quantitative results of the HBV cccDNA in each
cell according to a
fluorescence signal in each micro pore of the digital PCR detecting
chip. The digital PCR quantitative detecting kit for HBV cccDNA is strong in specificity, high in sensitivity, simple and convenient and quick. HBV cccDNA and HBV rcDNA are not needed to be separated and extracted, and the defects in the conventional HBV cccDNA detection are overcome, and therefore, the digital PCR quantitative detecting kit is suitable for large-scale popularization and application.