Establishment of fluorescent quantitative polymerase chain reaction detection method for human parvovirus B19

  Objective  To establish a fluorescent quantitative polymerase chain reaction detection method (PCR) method for simultaneous detection of human parvovirus B19(HPV B19).

  Methods  The online tool MAFFT was used to compare and analyze the sequence NS1 gene conserved region of three genotypes HPV B19, and specific primers and probes were designed for conserved region sequences. The fluorescent quantitative PCR method for detecting HPV B19 was established, and the sensitivity and specificity of the method were evaluated.

  Results  A standard curve drawn with a plasmid DNA template at a concentration of 1×10⁷ to 1×10² copies/μL and 20 copies/μL, the reaction efficiency E was 0.946 and R² was 0.996 6; the detection limit of this method was 10 copies/μL and the lower limit of quantification was 20 copies/μL. Herpes simplex virus (HSV) type 2, cytomegalovirus (CMV), group B streptococcus (GBS), hepatitis B virus (HBV), Epstein-Barr virus (EBV) positive DNA virus test results were negative, and the reference plasmid and HPV B19 positive samples were amplified.

  Conclusion  The established fluorescent quantitative PCR detection method can be applied for the screening and diagnosis of HPV B19 infection as high sensitivity and specificity.