Detection of HIV-1 drug resistance in women following administration of a single dose of nevirapine: comparison of plasma RNA to cellular DNA by consensus sequencing and by oligonucleotide ligation assay.
A single dose of nevirapine (sdNVP) to prevent mother-to-child transmission of HIV-1 increases the risk of failure of subsequent NVP-containing antiretroviral therapy (ART), especially when initiated within 6 months of sdNVP administration, emphasizing the importance of understanding the decay of nevirapine-resistant mutants. Nevirapine-resistant HIV-1 genotypes (with the mutations K103N, Y181C, and/or G190A) from 21 women were evaluated 10 days and 6 weeks after sdNVP administration and at the initiation of ART. Resistance was assayed by consensus sequencing and by a more sensitive assay (oligonucleotide ligation assay [OLA]) using plasma-derived HIV-1 RNA and cell-associated HIV-1 DNA. OLA detected nevirapine resistance in more specimens than consensus sequencing did (63% versus 33%, P<0.01). When resistance was detected only by OLA (n=45), the median mutant concentration was 18%, compared to 61% when detected by both sequencing and OLA (n=51) (P<0.0001). The proportion of women whose nevirapine resistance was detected by OLA 10 days after sdNVP administration was higher when we tested their HIV-1 RNA (95%) than when we tested their HIV-1 DNA (88%), whereas at 6 weeks after sdNVP therapy, the proportion was greater with DNA (85%) than with RNA (67%) and remained higher with DNA (33%) than with RNA (11%) at the initiation of antiretroviral treatment (median, 45 weeks after sdNVP therapy). Fourteen women started NVP-ART more than 6 months after sdNVP therapy; resistance was detected by OLA in 14% of the women but only in their DNA. HIV-1 resistance to NVP following sdNVP therapy persists longer in cellular DNA than in plasma RNA, as determined by a sensitive assay using sufficient copies of virus, suggesting that DNA may be superior to RNA for detecting resistance at the initiation of ART.
Wagner
Thor A
TA
University of Washington, Seattle Children's Hospital Research Institute, 1900 Ninth Ave, 8th Floor, Seattle, WA 98101, USA.
Kress
Catherine M
CM
Beck
Ingrid
I
Techapornroong
Malee
M
Wittayapraparat
Pakorn
P
Tansuphasawasdikul
Somboon
S
Jourdain
Gonzague
G
Ngo-Giang-Huong
Nicole
N
Lallemant
Marc
M
Frenkel
Lisa M
LM
D000328:Adult; D019380:Anti-HIV Agents / Q000627:therapeutic use*; D018890:Chemoprevention / Q000379:methods; D017931:DNA Primers / Q000235:genetics; D004279:DNA, Viral / Q000097:blood; D024882:Drug Resistance, Viral; D005260:Female; D015658:HIV Infections / Q000188:drug therapy; D015497:HIV-1 / Q000235:genetics* / Q000302:isolation & purification; D006801:Humans; D018445:Infectious Disease Transmission, Vertical / Q000517:prevention & control*; D022521:Ligase Chain Reaction / Q000379:methods; D008826:Microbial Sensitivity Tests / Q000379:methods; D019829:Nevirapine / Q000627:therapeutic use*; D015345:Oligonucleotide Probes / Q000235:genetics; D011247:Pregnancy; D011251:Pregnancy Complications, Infectious / Q000188:drug therapy; D012367:RNA, Viral / Q000097:blood; D012680:Sensitivity and Specificity; D017422:Sequence Analysis, DNA / Q000379:methods
D019380:Anti-HIV Agents; D017931:DNA Primers; D004279:DNA, Viral; D015345:Oligonucleotide Probes; D012367:RNA, Viral; D019829:Nevirapine