PCR detection of dengue virus using dried whole blood spotted on filter paper.
Whole blood dried onto filter paper constitutes a potentially useful material for molecular testing of viruses, including dengue. In order to assess the stability of viral RNA, we carried out dengue-RNA detection in whole blood infected with dengue virus that had been previously spotted onto filter paper. Filter papers were stored at room temperature, 4 and -70 degrees C and processed for PCR assay at intervals of 2, 4, 6 and 9 weeks. Our results demonstrated that dengue-RNA was stable in filter paper for 9 weeks at all tested temperatures. Furthermore, we evaluated these conditions using frozen sera and dried blood samples onto filter paper from 52 patients with confirmed clinical diagnosis of dengue infection. PCR results showed a 100% specificity and 93% sensitivity for dried blood samples. This storage method facilitates the transportation and analysis by nucleic acid amplification techniques even when freezing conditions are not available.
Prado
Irina
I
Virology Department, PAHO/WHO Collaborating Center for Viral Diseases, Pedro Kourí Tropical Medicine Institute, Autopista Novia del Mediodía, Km 6 1/2, P.O. Box Marianao 13, Havana, Cuba.
Rosario
Delfina
D
Bernardo
Lídice
L
Alvarez
Mayling
M
Rodríguez
Rosmari
R
Vázquez
Susana
S
Guzmán
María G
MG
D001769:Blood / Q000821:virology*; D001800:Blood Specimen Collection; D003715:Dengue / Q000175:diagnosis* / Q000821:virology; D003716:Dengue Virus / Q000235:genetics / Q000302:isolation & purification*; D006801:Humans; D010209:Paper; D016133:Polymerase Chain Reaction / Q000379:methods*; D020871:RNA Stability; D012367:RNA, Viral / Q000032:analysis* / Q000302:isolation & purification; D012680:Sensitivity and Specificity; D013696:Temperature; D013997:Time Factors
D012367:RNA, Viral