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Detection of dengue virus in saliva and urine by

real time RT-PCR


The electronic version of this article is the complete one and can be found online
at: http://www.virologyj.com/content/7/1/22
Received:

17 November 2009

Accepted:

27 January 2010

Published:

27 January 2010

2010 Poloni et al; licensee BioMed Central Ltd.


This is an Open Access article distributed under the terms of the Creative
Commons

Attribution

License

(http://creativecommons.org/licenses/by/2.0),

which permits unrestricted use, distribution, and reproduction in any medium,


provided the original work is properly cited.
Abstract
Early diagnosis of dengue virus (DENV) infection is important for patient
management and control of dengue outbreaks. The objective of this study was to
analyze the usefulness of urine and saliva samples for early diagnosis of DENV
infection by real time RT-PCR. Two febrile patients, who have been attended at
the General Hospital of the School of Medicine of Ribeirao Preto, Sao Paulo
University were included in the study. Serum, urine and saliva samples collected
from both patients were subjected to real time RT-PCR for DENV detection and
quantification. Dengue RNA was detected in serum, urine and saliva samples of
both patients. Patient 1 was infected with DENV-2 and patient 2 with DENV-3.
Data presented in this study suggest that urine and saliva could be used as
alternative samples for early diagnosis of dengue virus infection when blood

samples are difficult to obtain, e.g., in newborns and patients with hemorrhagic
syndromes.
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In addition to serum, we were able to detect dengue virus in both urine and saliva
samples by real time RT-PCR. In a previous case report study, dengue virus was
detected in urine and saliva samples up to 14 days after the onset of fever, when
no viremia was detected, suggesting that the virus could remain in urine and
saliva for a longer period of time than in serum [9] . However, in our study,
dengue virus was detected in serum, urine and saliva samples of both patients,
two days after the onset of symptoms, but not after nine days. The lower viral
load found in urine and saliva compared to serum suggests that dengue virus
would be detected in these clinical samples for a shorter period of time.
Recently, the usefulness of saliva in dengue serologic diagnosis has been
studied showing that specific IgM could be detected in patients with primary
infection and IgM/IgG in patients with secondary infection [10] . We were able to
detect IgM and IgG in the two patients with a primary infection.
Our results suggest that urine and saliva samples could be used for dengue
diagnosis in cases where blood samples are difficult to obtain, such as in
newborns or in patients with hemorrhagic syndromes. Urine and saliva samples
are easy to collect without invasive procedures and urine samples can be

collected in large volumes. However, further studies, with a higher number of


patients, are needed to confirm these results.
List of abbreviations
(DENV): Dengue virus; (DF): Dengue fever; (DHF/DSS): Dengue hemorrhagic
fever/dengue shock syndrome; (RT-PCR): Reverse transcription-polymerase
chain reaction; (NJ): Neighbor-joining; (hLTR): Hierarchical likelihood ratio test.

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