Viscoelastic Separation and Concentration of Fungi from Blood for Highly Sensitive Molecular Diagnostics

Jeonghun Nam, Woong Sik Jang, Da Hye Hong, Chae Seung Lim

Research output: Contribution to journalArticle

2 Citations (Scopus)

Abstract

Isolation and concentration of fungi in the blood improves sensitivity of the polymerase chain reaction (PCR) method to detect fungi in blood. This study demonstrates a sheathless, continuous separation and concentration method of candida cells using a viscoelastic fluid that enables rapid detection of rare candida cells by PCR analysis. To validate device performance using a viscoelastic fluid, flow characteristics of 2 μm particles were estimated at different flow rates. Additionally, a mixture of 2 μm and 13 μm particles was successfully separated based on size difference at 100 μl/min. Candida cells were successfully separated from the white blood cells (WBCs) with a separation efficiency of 99.1% and concentrated approximately 9.9-fold at the center outlet compared to the initial concentration (~2.5 × 10 7 cells/ml). Sequential 1st and 2nd concentration processes were used to increase the final number of candida cells to ~2.3 × 10 9 cells/ml, which was concentrated ~92-fold. Finally, despite the undetectable initial concentration of 10 1 CFU/ml, removal of WBCs and the additional buffer solution enabled the quantitative reverse transcription (RT)-PCR detection of candida cells after the 1st concentration (Ct = 31.43) and the 2nd concentration process (Ct = 29.30).

Original languageEnglish
Article number3067
JournalScientific Reports
Volume9
Issue number1
DOIs
Publication statusPublished - 2019 Dec 1

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Molecular Pathology
Candida
Fungi
Polymerase Chain Reaction
Leukocytes
Reverse Transcription
Buffers
Cell Count
Equipment and Supplies

ASJC Scopus subject areas

  • General

Cite this

Viscoelastic Separation and Concentration of Fungi from Blood for Highly Sensitive Molecular Diagnostics. / Nam, Jeonghun; Jang, Woong Sik; Hong, Da Hye; Lim, Chae Seung.

In: Scientific Reports, Vol. 9, No. 1, 3067, 01.12.2019.

Research output: Contribution to journalArticle

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