Improved detection of HIV-2 proviral DNA in dually seroreactive individuals by PCR.
OBJECTIVE
To improve the detection rate of HIV-2 proviral DNA in primary uncultured peripheral blood mononuclear cells (PBMC) of HIV-2-seroreactive and HIV-1-HIV-2 dually seroreactive individuals.


METHODS
Two newly designed HIV-2 PCR primer pairs in the long terminal repeat (LTR) gag and gag-pol regions and a previously described env and LTR HIV-2 PCR primer pairs were tested on samples from 66 confirmed HIV-2-seropositive individuals (The Gambia, 40; Côte d'Ivoire, 17; Guinea-Bissau, nine), 209 dually seroreactive individuals (The Gambia, 82; Côte d'Ivoire, 127), 24 genetically characterized isolated HIV-1 strains (group M subtypes A-H and group O), one simian immunodeficiency virus (SIV) strain cpz, 10 HIV-2 isolates (subtype A, B and unidentified), two SIVsm isolates, and 10 seronegative samples.


RESULTS
All HIV-2 primers evaluated showed 100% specificity since there was no amplification observed with 24 HIV-1, one SIVcpz and 10 seronegative samples. One single copy of the HIV-2 genome could be detected with all outer primer pairs as well as all inner primer pairs on one PCR round used. Sensitivity of primers (at least one of the four primer pairs was positive) to HIV-2-seropositive samples was 100% (all nine) in Guinea-Bissau, 71% (12/17) in Côte d'Ivoire, 100% (all 20) in Gambian AIDS patients, and 85% (17/20) in Gambian pregnant women. Doubling the PBMC of dually seroreactive individuals from 7.5 x 10(4) to 1.5 x 10(5) in the PCR revealed the presence of both HIV-1 and 2 proviral DNA in 72% (92/127) in Côte d'Ivoire and 72% (59/82) in The Gambia. By doubling the number of PBMC, HIV-2 detection in dually seroreactive individuals by PCR was increased from 65 to 77% in Côte d'Ivoire and from 67 to 83% in The Gambia.


CONCLUSIONS
The use of 1.5 x 10(5) primary uncultured PBMC and the newly designed HIV-2 primer pairs allowed us to document the highest percentage (72%) ever reported of HIV-1-HIV-2 dual infections amongst HIV-1-HIV-2 dually seroreactive individuals in Côte d'Ivoire and The Gambia. Improved detection of HIV-2 proviral DNA, rather than exposure to both viruses, infection with only one virus, or infection with a unique third virus containing epitopes common to both HIV-1 and HIV-2, contributes to a more accurate monitoring of the prevalence of HIV-1-HIV-2 dual infections.
Ishikawa
K
K
Institute of Tropical Medicine, Antwerp, Belgium.
Fransen
K
K
Ariyoshi
K
K
Nkengasong
J N
JN
Janssens
W
W
Heyndrickx
L
L
Whittle
H
H
Diallo
M O
MO
Ghys
P D
PD
Coulibaly
I M
IM
Greenberg
A E
AE
Piedade
J
J
Canas-Ferreira
W
W
van der Groen
G
G
D017931:DNA Primers; D004279:DNA, Viral / Q000032:analysis*; D005260:Female; D005714:Gambia; D015752:Genes, env; D015750:Genes, gag; D015751:Genes, pol; D016325:HIV Long Terminal Repeat; D006679:HIV Seropositivity / Q000175:diagnosis* / Q000276:immunology / Q000821:virology*; D015497:HIV-1; D015498:HIV-2; D006801:Humans; D016133:Polymerase Chain Reaction / Q000379:methods*; D011247:Pregnancy; D011251:Pregnancy Complications, Infectious / Q000821:virology*; D011533:Proviruses; D012680:Sensitivity and Specificity
D017931:DNA Primers; D004279:DNA, Viral