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					Author(s): Huang, SH (Huang, Su-Hua); Yang, TC (Yang, Tsuey-Ching); Tsai, MH (Tsai,
Ming-Hong); Tsai, IS (Tsai, I-Shou); Lu, HC (Lu, Huang-Chih); Chuang, PH (Chuang,
Pei-Hsin); Wan, L (Wan, Lei); Lin, YJ (Lin, Ying-Ju); Lai, CH (Lai, Chih-Ho); Lin, CW (Lin,
Cheng-Wen)

Title: Gold nanoparticle-based RT-PCR and real-time quantitative RT-PCR assays for
detection of Japanese encephalitis virus

Source: NANOTECHNOLOGY, 19 (40): Art. No. 405101 OCT 8 2008

Language: English

Document Type: Article

KeyWords Plus: POLYMERASE-CHAIN-REACTION; ELECTRON DYNAMICS; SIZE; DNA;
AMPLIFICATION; DIAGNOSIS; RELAXATION; PARTICLES

Abstract: Virus isolation and antibody detection are routinely used for diagnosis of Japanese
encephalitis virus (JEV) infection, but the low level of transient viremia in some JE patients
makes JEV isolation from clinical and surveillance samples very difficult. We describe the use
of gold nanoparticle-based RT-PCR and real-time quantitative RT-PCR assays for detection of
JEV from its RNA genome. We tested the effect of gold nanoparticles on four different PCR
systems, including conventional PCR, reverse-transcription PCR (RT-PCR), and SYBR green
real-time PCR and RT-PCR assays for diagnosis in the acute phase of JEV infection. Gold
nanoparticles increased the amplification yield of the PCR product and shortened the PCR
time compared to the conventional reaction. In addition, nanogold-based real-time RT-PCR
showed a linear relationship between Ct and template amount using ten-fold dilutions of JEV.
The nanogold-based RT-PCR and real-time quantitative RT-PCR assays were able to detect
low levels (1-10 000 copies) of the JEV RNA genomes extracted from culture medium or whole
blood, providing early diagnostic tools for the detection of low-level viremia in the acute-phase
infection. The assays described here were simple, sensitive, and rapid approaches for
detection and quantitation of JEV in tissue cultured samples as well as clinical samples.

Addresses: [Yang, Tsuey-Ching; Tsai, Ming-Hong; Chuang, Pei-Hsin; Lin, Cheng-Wen]
China Med Univ, Dept Med Lab Sci & Biotechnol, Taichung 404, Taiwan; [Huang, Su-Hua;
Tsai, Ming-Hong; Lin, Cheng-Wen] Asia Univ, Dept Biotechnol, Coll Hlth Sci, Taichung,
Taiwan; [Tsai, I-Shou; Lu, Huang-Chih] Feng Chia Univ, Nanotechnol Res Ctr, Taichung
40724, Taiwan; [Wan, Lei; Lin, Ying-Ju] China Med Univ Hosp, Dept Med Genet & Med Res,
Taichung, Taiwan; [Lai, Chih-Ho] China Med Univ, Dept Microbiol & Immunol, Taichung,
Taiwan; [Lin, Cheng-Wen] China Med Univ Hosp, Clin Virol Lab, Dept Lab Med, Taichung,
Taiwan

Reprint Address: Lin, CW, China Med Univ, Dept Med Lab Sci & Biotechnol, 91 Hsueh Shih
Rd, Taichung 404, Taiwan.

E-mail Address: cwlin@mail.cmu.edu.tw
Funding Acknowledgement:

Funding Agency                      Grant Number

National Science Council (Taiwan)

                                    NSC96-2320- B-039-008-MY3
China Medical University
                                    CMU95-070

We would like to thank the National Science Council (Taiwan) and China Medical University
for financial support (NSC96-2320- B-039-008-MY3 and CMU95-070).

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Cited Reference Count: 28
Times Cited: 2
Publisher: IOP PUBLISHING LTD
Publisher Address: DIRAC HOUSE, TEMPLE BACK, BRISTOL BS1 6BE, ENGLAND
ISSN: 0957-4484
Article Number: 405101
DOI: 10.1088/0957-4484/19/40/405101
29-char Source Abbrev.: NANOTECHNOL
ISO Source Abbrev.: Nanotechnology
Source Item Page Count: 8
Subject Category: Engineering, Multidisciplinary; Nanoscience & Nanotechnology; Materials
Science, Multidisciplinary; Physics, Applied
ISI Document Delivery No.: 341TI

				
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