𝔖 Bobbio Scriptorium
✦   LIBER   ✦

Development and evaluation of a fluorogenic real-time RT-PCR for the detection of dengue 3 virus

✍ Scribed by Irene L. Tan; Mark Pierre S. Dimamay; Corazon C. Buerano; Jhoe Anthony R. Alfon; Carol Z. Tanig; Ronald R. Matias; Filipinas F. Natividad

Publisher
John Wiley and Sons
Year
2010
Tongue
English
Weight
297 KB
Volume
82
Category
Article
ISSN
0146-6615

No coin nor oath required. For personal study only.

✦ Synopsis

Abstract

A dengue‐3‐specific real‐time reverse transcriptase‐polymerase chain reaction (RT‐PCR) was developed using the novel __L__ight __U__pon e__X__tension (LUX™) fluorogenic technology. A labeled forward primer and a standard reverse primer that target a conserved region within the non‐structural 1 (NS1) gene of dengue 3 strains were designed. The dengue‐3‐specific assay did not recognize other dengue serotypes and related flaviviruses. Using a tenfold serial dilution of plasmid DNA containing the dengue 3 NS1 gene as standards, the range of dengue virus detection was determined to be from 10^3^ to 10^9^ copies/ml or from 80 to 8 × 10^7^ copies/reaction with an average correlation coefficient of ≥0.99. The mean intra‐assay coefficient of variation (CV) at 2.01% and the mean inter‐assay CV at 2.68% suggest the repeatability of the procedure. Moreover, the fluorogenic assay was evaluated by using clinical specimens and comparing test results with historical data obtained from conventional RT‐PCR, which served as the criterion standard. Using patient sera as test samples, the assay demonstrated 95.45% sensitivity and 100% specificity, respectively. These results reveal that the real‐time RT‐PCR assay may be utilized as a rapid, convenient, and sensitive tool for the detection of dengue 3 in clinical and laboratory specimens. J. Med. Virol. 82:2053–2063, 2010. © 2010 Wiley‐Liss, Inc.

📜 SIMILAR VOLUMES

A simple one-step real-time RT-PCR for d
A simple one-step real-time RT-PCR for diagnosis of dengue virus infection
✍ Harryson Wings Godoy dos Santos; Telma Regina Ramos Silva Poloni; Kelly Paula So 📂 Article 📅 2008 🏛 John Wiley and Sons 🌐 English ⚖ 126 KB 👁 1 views

## Abstract Dengue is the most important arbovirus disease in tropical and sub‐tropical countries, and can be caused by infection with any of the four‐dengue virus (DENV) serotypes. Infection with DENV can lead to a broad clinical spectrum, ranging from sub‐clinical infection or an influenza‐like d

Comparison of real-time fluorescent RT-P
Comparison of real-time fluorescent RT-PCR and conventional RT-PCR for the detection of hepatitis E virus genotypes prevalent in China
✍ Chenyan Zhao; Zhuo Li; Baoshan Yan; Tim J. Harrison; Xinhui Guo; Feng Zhang; Jim 📂 Article 📅 2007 🏛 John Wiley and Sons 🌐 English ⚖ 128 KB 👁 1 views

## Abstract To compare the specificity and sensitivity of a real‐time fluorescent RT‐PCR assay with conventional RT‐PCR, sera from 110 healthy blood donors, 120 patients with a clinical diagnosis of chronic hepatitis B, and 416 patients with non‐A‐C acute hepatitis, as well as serial dilutions of H

Development and evaluation of a ‘real-ti
Development and evaluation of a ‘real-time’ RT-PCR for the detection of enterovirus and parechovirus RNA in CSF and throat swab samples
✍ Caroline E. Corless; Malcolm Guiver; Raymond Borrow; Valerie Edwards-Jones; Andr 📂 Article 📅 2002 🏛 John Wiley and Sons 🌐 English ⚖ 117 KB 👁 1 views

## Abstract A two‐step reverse transcriptase TaqMan™ duplex PCR (RT‐PCR) assay was developed using the ABI 7700 Sequence Detection System for the detection of enterovirus (EV) and parechovirus type 1 and 2 (PEV) RNA from samples of cerebrospinal fluid (CSF) and throat swabs. Using sequence‐specific

Detection and characterization of norovi
Detection and characterization of norovirus outbreaks in Germany: Application of a one-tube RT-PCR using a fluorogenic real-time detection system
✍ M. Höhne; E. Schreier 📂 Article 📅 2003 🏛 John Wiley and Sons 🌐 English ⚖ 259 KB

## Abstract Outbreaks of gastroenteritis caused by Norovirus are an increasing public health problem worldwide. The virus is transmitted by contaminated food and by person‐to‐person. In Germany, a new health reporting system including Norovirus infections has been introduced in 2001. Norovirus outb

Comparative evaluation of the seegene se
Comparative evaluation of the seegene seeplex RV15 and real-time PCR for respiratory virus detection
✍ David F. Bibby; Iain McElarney; Judith Breuer; Duncan A. Clark 📂 Article 📅 2011 🏛 John Wiley and Sons 🌐 English ⚖ 118 KB 👁 1 views

## Abstract Respiratory virus infections contribute substantially both to hospitalizations of young children, and to morbidity in immunocompromised patients such as those with hematological malignancies. Their rapid and accurate diagnosis is essential to patient management. To evaluate the prospect

Development and evaluation of a real-tim
Development and evaluation of a real-time PCR assay for rapid identification and semi-quantitation of measles virus
✍ Brenda Thomas; Stuart Beard; Li Jin; Kevin E. Brown; David W.G. Brown 📂 Article 📅 2007 🏛 John Wiley and Sons 🌐 English ⚖ 111 KB 👁 1 views

## Abstract A real‐time PCR assay for measles virus was designed and validated using clinical samples including oral fluids, sera, urines, throat swabs, blood samples, and nasopharyngeal aspirates. The test was specific for measles virus, with a slightly higher sensitivity compared to the conventio