Publication: Application of one-step reverse transcription droplet digital pcr for dengue virus detection and quantification in clinical specimens
4
Issued Date
2021-04-01
Resource Type
ISSN
20754418
Other identifier(s)
2-s2.0-85108986648
Rights
Mahidol University
Rights Holder(s)
SCOPUS
Bibliographic Citation
Diagnostics. Vol.11, No.4 (2021)
Suggested Citation
Dumrong Mairiang, Adisak Songjaeng, Prachya Hansuealueang, Yuwares Malila, Paphavee Lertsethtakarn, Sasikorn Silapong, Yongyuth Poolpanichupatam, Chonticha Klungthong, Kwanrutai Chin-Inmanu, Somchai Thiemmeca, Nattaya Tangthawornchaikul, Kanokwan Sriraksa, Wannee Limpitikul, Sirijitt Vasanawathana, Damon W. Ellison, Prida Malasit, Prapat Suriyaphol, Panisadee Avirutnan Application of one-step reverse transcription droplet digital pcr for dengue virus detection and quantification in clinical specimens. Diagnostics. Vol.11, No.4 (2021). doi:10.3390/diagnostics11040639 Retrieved from: https://repository.li.mahidol.ac.th/handle/123456789/76216
Research Projects
Organizational Units
Authors
Journal Issue
Thesis
Title
Application of one-step reverse transcription droplet digital pcr for dengue virus detection and quantification in clinical specimens
Author(s)
Dumrong Mairiang
Adisak Songjaeng
Prachya Hansuealueang
Yuwares Malila
Paphavee Lertsethtakarn
Sasikorn Silapong
Yongyuth Poolpanichupatam
Chonticha Klungthong
Kwanrutai Chin-Inmanu
Somchai Thiemmeca
Nattaya Tangthawornchaikul
Kanokwan Sriraksa
Wannee Limpitikul
Sirijitt Vasanawathana
Damon W. Ellison
Prida Malasit
Prapat Suriyaphol
Panisadee Avirutnan
Adisak Songjaeng
Prachya Hansuealueang
Yuwares Malila
Paphavee Lertsethtakarn
Sasikorn Silapong
Yongyuth Poolpanichupatam
Chonticha Klungthong
Kwanrutai Chin-Inmanu
Somchai Thiemmeca
Nattaya Tangthawornchaikul
Kanokwan Sriraksa
Wannee Limpitikul
Sirijitt Vasanawathana
Damon W. Ellison
Prida Malasit
Prapat Suriyaphol
Panisadee Avirutnan
Abstract
Detection and quantification of viruses in laboratory and clinical samples are standard assays in dengue virus (DENV) studies. The quantitative reverse transcription polymerase chain reaction (qRT-PCR) is considered to be the standard for DENV detection and quantification due to its high sensitivity. However, qRT-PCR offers only quantification relative to a standard curve and consists of several “in-house” components resulting in interlaboratory variations. We developed and optimized a protocol for applying one-step RT-droplet digital PCR (RT-ddPCR) for DENV detection and quantification. The lower limit of detection (LLOD95) and the lower limit of quantification (LLOQ) for RT-ddPCR were estimated to be 1.851 log10-copies/reaction and 2.337 log10-copies/reaction, respectively. The sensitivity of RT-ddPCR was found to be superior to qRT-PCR (94.87% vs. 90.38%, p = 0.039) while no false positives were detected. Quantification of DENV in clinical samples was independently performed in three laboratories showing interlaboratory variations with biases <0.5 log10-copies/mL. The RT-ddPCR protocol presented here could help harmonize DENV quantifi cation results and improve findings in the field such as identifying a DENV titer threshold correlating with disease severity.