Publication:
Comparison of droplet digital PCR and real time PCR method for HBV DNA quantification

1
0

Issued Date

2015

Resource Type

Article

File Type

application/pdf

ISSN

1252208

Other identifier(s)

2-s2.0-84957625468

Rights Holder(s)

Scopus

Bibliographic Citation

Journal of the Medical Association of Thailand. Vol 98, (2015), p.S140-S145

Suggested Citation

Wongjitrat C., Horthongkham N., Sutthent R., Srisurapanon S. Comparison of droplet digital PCR and real time PCR method for HBV DNA quantification. Journal of the Medical Association of Thailand. Vol 98, (2015), p.S140-S145. Retrieved from: https://hdl.handle.net/20.500.14740/6075

Title

Comparison of droplet digital PCR and real time PCR method for HBV DNA quantification

Author(s)

Wongjitrat C.
 Horthongkham N.
 Sutthent R.
 Srisurapanon S.

Abstract

Background: HBV infection causes a potential serious public health problem. The ability to detect HBV DNA concentration is an important issue that had been continuously improved. When using quantitative polymerase chain reaction (qPCR), several factors are of concern, for example, sources of material, standard curve calibration, and PCR efficiency. Digital PCR (dPCR) is an alternative PCR-based technique for absolute quantification using Poisson’s statistics without requiring a standard curve. Objective: Compare the data set of HBV DNA generated between dPCR and qPCR methods. Material and Methods: Fifty-four samples were quantified by Abbot’s real time PCR and with 2-6 log10 HBV DNA were selected for comparison with dPCR. Results: Of these 54 samples, there were two outlier samples defined as negative by dPCR, whereas 52 samples were positive by both of these assays. The difference between two assays was less than 0.25 log IU/mL in 24/52 samples (46%) of paired samples; less than 0.5 log IU/mL in 46/52 samples (88%) and less than 1 log in 50/52 samples (96%). The correlation coefficient (r) was 0.788 (p-value <0.0001). Comparison with qPCR method, data generated by dPCR tend to be an overestimation in the sample with the low level of HBV DNA concentration and underestimated in the sample with high viral load. The variation of DNA by dPCR measurement might be due to the pre-amplification procedure and PCR template. Conclusion: Measurement of HBV DNA by using dPCR, the results of the HBV DNA copy number tended to be deviated by over- or under-estimated when comparison to real time PCR method. In addition, a large quantity of DNA was used when compared to qPCR. However, the optimum processes of this assay have to be further investigated. © 2015, Medical Association of Thailand. All rights reserved.

Subject(s)

Correlation coefficient
 Gene amplification
 Intermethod comparison
 Major clinical study
 Polymerase chain reaction
 Quantitative study
 Real time polymerase chain reaction
 Statistical significance
 Statistics
 Virus load
 Blood
 Calibration
 Comparative study
 Hepatitis B
 Human
 Polymerase chain reaction
 Procedures
 Real time polymerase chain reaction
 Virus DNA
 Calibration
 DNA, Viral
 Hepatitis B
 Humans
 Polymerase Chain Reaction
 Real-Time Polymerase Chain Reaction
 Viral Load

View online Resources

Research Projects

Organizational Units

Journal Issue

URI

https://hdl.handle.net/20.500.14740/6075

Collections

Scopus: Year 1983-2021

Endorsement

Review

Supplemented By

Referenced By