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

XU Liangyun; ZHANG Qigang; FAN Guanglai; ZHAO Jianhua

doi:10.7619/jcmp.20210246

Journal of Clinical Medicine in Practice > 2021 > 25(7): 11-14, 20. > DOI: 10.7619/jcmp.20210246

XU Liangyun, ZHANG Qigang, FAN Guanglai, ZHAO Jianhua. Establishment of fluorescent quantitative polymerase chain reaction detection method for human parvovirus B19[J]. Journal of Clinical Medicine in Practice, 2021, 25(7): 11-14, 20. DOI: 10.7619/jcmp.20210246

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

1.
Tumor Hospital Affiliated to Fourth Clinical Medical College of Nanjing Medical University, Nanjing, Jiangsu, 210009
2.
Gene Testing Technology Application Demonstration Center, Huai'an City Maternal and Child Health Hospital of Jiangsu Province, Huai'an, Jiangsu, 223002

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Received Date: January 12, 2021
Available Online: April 20, 2021
Published Date: July 27, 2021

Graphical Abstract

Abstract

Abstract

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.
- human parvovirus B19,
- fluorescent quantitative polymerase chain reaction,
- standard curve,
- sequence

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References

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