Dengue virus detection using whole blood for reverse transcriptase PCR and virus isolation.
Dengue is one of the most important diseases in the tropical and subtropical regions of the world, with an estimated 2.5 billion people being at risk. Detection of dengue virus infections has great importance for the clinical management of patients, surveillance, and clinical trial assessments. Traditionally, blood samples are collected in serum separator tubes, processed for serum, and then taken to the laboratory for analysis. The use of whole blood has the potential advantages of requiring less blood, providing quicker results, and perhaps providing better sensitivity during the acute phase of the disease. We compared the results obtained by reverse transcriptase PCR (RT-PCR) with blood drawn into tubes containing EDTA and those obtained by RT-PCR with blood samples in serum separator tubes from 108 individuals clinically suspected of being infected with dengue virus. We determined that the extraction of RNA from whole blood followed by RT-PCR resulted in a higher detection rate than the use of serum or plasma. Using a selection of these samples, we also found that our ability to detect virus by direct C6/36 cell culture and mosquito inoculation was enhanced by using whole blood but not to the same extent as that seen by the use of RT-PCR.
Klungthong
Chonticha
C
Armed Forces Research Institute of Medical Sciences, Department of Virology, Bangkok, Thailand.
Gibbons
Robert V
RV
Thaisomboonsuk
Butsaya
B
Nisalak
Ananda
A
Kalayanarooj
Siripen
S
Thirawuth
Vipa
V
Nutkumhang
Naowayubol
N
Mammen
Mammen P
MP
Jarman
Richard G
RG
D001769:Blood / Q000821:virology*; D002648:Child; D003715:Dengue / Q000821:virology; D003716:Dengue Virus / Q000302:isolation & purification*; D005260:Female; D006801:Humans; D008297:Male; D010949:Plasma / Q000821:virology; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity; D044967:Serum / Q000821:virology; D013048:Specimen Handling / Q000379:methods*; D014776:Virus Cultivation / Q000379:methods