TY - JOUR
T1 - Improved detection of HIV-2 proviral DNA in dually seroreactive individuals by PCR
AU - Ishikawa, Koichi
AU - Fransen, Katrien
AU - Ariyoshi, Koya
AU - Nkengasong, John N.
AU - Janssens, Wouter
AU - Heyndrickx, Leo
AU - Whittle, Hilton
AU - Diallo, Mamadou O.
AU - Ghys, Peter D.
AU - Coulibaly, Issa Malick
AU - Greenberg, Alan E.
AU - Piedade, João
AU - Canas-Ferreira, Wanda
AU - Van Der Groen, Guido
PY - 1998/8/20
Y1 - 1998/8/20
N2 - 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. Materials and 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; Cote d'lvoire, 17, Guinea-Bissau, nine), 209 dually seroreactive individuals (The Gambia, 82; Cote d'lvoire, 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 SIV(sm) 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 SIV(cpz) 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 Cote d'lvoire, 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 104 to 1.5 x 105 in the PCR revealed the presence of both HIV-1 and 2 proviral DNA in 72% (92/127) in Cote d'lvoire 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 Cote d'lvoire and from 67 to 83% in The Gambia. Conclusions: The use of 1.5 x 105 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 Cote d'lvoire 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.
AB - 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. Materials and 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; Cote d'lvoire, 17, Guinea-Bissau, nine), 209 dually seroreactive individuals (The Gambia, 82; Cote d'lvoire, 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 SIV(sm) 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 SIV(cpz) 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 Cote d'lvoire, 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 104 to 1.5 x 105 in the PCR revealed the presence of both HIV-1 and 2 proviral DNA in 72% (92/127) in Cote d'lvoire 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 Cote d'lvoire and from 67 to 83% in The Gambia. Conclusions: The use of 1.5 x 105 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 Cote d'lvoire 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.
KW - Dual seroreactivity
KW - HIV-2
KW - PCR
KW - West Africa
UR - http://www.scopus.com/inward/record.url?scp=15144357227&partnerID=8YFLogxK
UR - https://journals.lww.com/aidsonline/Fulltext/1998/12000/Improved_detection_of_HIV_2_proviral_DNA_in_dually.3.aspx
U2 - 10.1097/00002030-199812000-00003
DO - 10.1097/00002030-199812000-00003
M3 - Article
C2 - 9727562
AN - SCOPUS:15144357227
VL - Vol. 12
SP - 1419
EP - 1425
JO - AIDS
JF - AIDS
SN - 0269-9370
IS - n.º 12
ER -