Journal of
Infectious Diseases and Immunity

  • Abbreviation: J. Infect. Dis. Immun.
  • Language: English
  • ISSN: 2141-2375
  • DOI: 10.5897/JIDI
  • Start Year: 2009
  • Published Articles: 87

Full Length Research Paper

Virological diagnosis of dengue fever in Jeddah, Saudi Arabia: Comparison between RT-PCR and virus isolation in cell culture

Esam Azhar1*, Moujahed Kao1, Matthias Niedrig2, Badr Masri1, Azad Godus1, Raed Badierah1, Nassem Khan3, Adnan Almazrooa4, Ahmad Ashshi5 and Ghazi Jamjoom1
1Special Infectious Agents Unit, Bio-safety level - 3, King Fahd Medical Research Center, King Abdulaziz University, P. O. Box: 128442, Jeddah-21362, Saudi Arabia. 2Robert Koch-Institute, Viral Hemorrhagic Fever Department, Nordufer 20, 13355 Berlin /Germany. 3Division of Rheumatology, University of Arkansas for Medical Sciences, Mail Slot - 509, 4301 West Markham, Little Rock, AR 72205, USA. 4Faculty of Medicine, King Abdulaziz University, Jeddah, Saudi Arabia. 5Medical Laboratory Department, Faculty of Applied Medical Sciences, Umm Al-Qura University, Makkah, Saudi Arabia.
Email: [email protected]

  •  Accepted: 27 September 2010
  •  Published: 30 November 2010

Abstract

A total of 233 serum samples were collected from patients presenting to King Abdulaziz University Hospital with suspected cases of Dengue Fever (DF) from 2006 to 2008. Dengue virus was successfully isolated from 70 samples by culture on C6/36 and LLC-MK2 cells; it was then detected by indirect immunofluorescence assay (IFA). The cytopathic effect (CPE) of dengue virus on C6/36 appeared in most of the samples within 1-4 days post-inoculation comparing to 7-12 days on LLC-MK2 cells, and this was characterized by the ability to induce syncytia and multinucleated giant cells.  On the other hand, by using RT-PCR technique, 87 (37.3%) samples were positive. All 70 (30.4%) samples with positive cell culture results were detectable by RT-PCR in addition to 17 culture-negative samples were RT-PCR positive. Dengue virus type 1 (DENV-1) was the dominant serotype followed by DENV- 3 and DENV-2, while DENV-4 was not detected in tested samples. These results indicate that DENV-RNA detection by RT-PCR is more sensitive than virus isolation. We suggest that the high sensitivity coupled with the turnaround time, have made the RT-PCR a better choice as a routine test for DENV diagnosis.

 

Key words: Dengue virus, Dengue fever, RT-PCR, IIF, Saudi Arabia, Jeddah, Dengue diagnosis, virus isolation, virus replication, C6/36, LLCMK-2.