Usage of dried blood spots for molecular diagnosis and monitoring HIV-1 infection.
The usage of dried blood spots as specimens for diagnosis and monitoring of HIV-1 infection in Thailand was evaluated. EDTA blood samples, which were collected from 100 HIV seronegative and 109 HIV seropositive individuals, were tested on dried blood spots; Whatman, Schleicher and Schuell (S&S) No. 903 and S&S IsoCode filter paper. Nucleic acid was extracted and used as a template for HIV-1 proviral DNA detection by an "in-house" multiplex PCR and a commercial Amplicor HIV-1 PCR test (Roche, version 1.0). HIV-1 RNA qualitative (QL) and quantitative (QT) detection was determined by Nucleic Acid Sequence Based Amplification (NASBA). The average DNA per blood spot recovered from Whatman and S&S IsoCode was not statistically different (p = 0.512) with a range of 218.9+/-46.84 and 225.63+/-88.33 microg, respectively. The concordance of HIV-1 proviral DNA detection by PCR from dried blood spots Whatman and S&S IsoCode was 94% versus 89.4% for sensitivity and 100% versus 100% for specificity. The sensitivity and specificity of HIV-1 RNA QL detection in dried blood spots was 89.7 and 97.5%, respectively. The HIV-1 RNA QT from dried blood spots showed a good correlation in paired dried blood spots and plasma with Pearson correlation, r = 0.817 (R2 = 0.667, P < 0.05). The data showed that dried blood spots could be used for the diagnosis and monitoring of HIV-1 infection.
Uttayamakul
Sumonmal
S
National HIV Repository and Bioinformatic Center, Department of Microbiology, Faculty of Medicine Siriraj Hospital, Mahidol University, 2 Prannok Road, Bangkok 10700, Thailand.
Likanonsakul
Sirirat
S
Sunthornkachit
Rujanee
R
Kuntiranont
Karoon
K
Louisirirotchanakul
Suda
S
Chaovavanich
Achara
A
Thiamchai
Vipa
V
Tanprasertsuk
Sombat
S
Sutthent
Ruengpung
R
D001769:Blood / Q000821:virology; D004279:DNA, Viral / Q000032:analysis / Q000302:isolation & purification; D005260:Female; D015658:HIV Infections / Q000175:diagnosis* / Q000635:transmission / Q000821:virology; D015497:HIV-1 / Q000235:genetics / Q000302:isolation & purification*; D006801:Humans; D007223:Infant; D007231:Infant, Newborn; D018445:Infectious Disease Transmission, Vertical; D016133:Polymerase Chain Reaction; D011247:Pregnancy; D011251:Pregnancy Complications, Infectious / Q000175:diagnosis / Q000821:virology; D011533:Proviruses / Q000302:isolation & purification; D012367:RNA, Viral / Q000032:analysis / Q000302:isolation & purification; D011933:Reagent Kits, Diagnostic; D021161:Self-Sustained Sequence Replication; D012680:Sensitivity and Specificity; D013785:Thailand
D004279:DNA, Viral; D012367:RNA, Viral; D011933:Reagent Kits, Diagnostic