Comparison of LigAmp and an ASPCR assay for detection and quantification of K103N-containing HIV variants

Jessica D. Church, William I. Towler, Donald R. Hoover, Sarah E. Hudelson, Newton Kumwenda, Taha E Taha, James Eshleman, Susan Eshleman

Research output: Contribution to journalArticle

Abstract

We compared the ability of the LigAmp assay and an ASPCR assay to detect and quantify K103N-containing HIV variants in samples from 63 women who received single-dose nevirapine in a clinical trial. Samples were first analyzed with the ViroSeq HIV Genotyping system, and ViroSeq PCR products were used as templates for the LigAmp and ASPCR assays. A cutoff of 0.5% K103N for detection of K103N was used for both assays. Results for the percentage K103N were similar for the two assays (R2 = 0.92). Forty-six samples (73.0%) were positive for K103N by both assays and 13 samples (20.6%) were negative by both assays. Four samples (6.3%) were positive by ASPCR only. No samples were positive by LigAmp only. Eight discordant samples were analyzed in more detail. Sequence polymorphisms near oligonucleotide binding sites provided a possible explanation for the discordance in four of eight samples. The percentage K103N was also determined by analyzing 40 HIV clones from each of these eight samples, using a combined amplification/sequencing method (AmpliSeq). The percentage K103N determined by clonal analysis was consistent with the LigAmp result for five of eight samples, and was consistent with the ASPCR result for three of eight samples. Among 320 clones analyzed, we identified eight different codons at position 103 (mean = 3.8 codons/sample), which encoded six different amino acids, illustrating the extensive genetic diversity in HIV. Further studies are needed to compare performance of assays for detection and quantification of HIV drug resistance mutations in clinical samples.

Original languageEnglish (US)
Pages (from-to)595-605
Number of pages11
JournalAIDS Research and Human Retroviruses
Volume24
Issue number4
DOIs
StatePublished - Apr 1 2008

Fingerprint

HIV
Codon
Clone Cells
Nevirapine
Drug Resistance
Oligonucleotides
Binding Sites
Clinical Trials
Amino Acids
Polymerase Chain Reaction
Mutation

ASJC Scopus subject areas

  • Immunology
  • Virology

Cite this

Comparison of LigAmp and an ASPCR assay for detection and quantification of K103N-containing HIV variants. / Church, Jessica D.; Towler, William I.; Hoover, Donald R.; Hudelson, Sarah E.; Kumwenda, Newton; Taha, Taha E; Eshleman, James; Eshleman, Susan.

In: AIDS Research and Human Retroviruses, Vol. 24, No. 4, 01.04.2008, p. 595-605.

Research output: Contribution to journalArticle

Church, Jessica D. ; Towler, William I. ; Hoover, Donald R. ; Hudelson, Sarah E. ; Kumwenda, Newton ; Taha, Taha E ; Eshleman, James ; Eshleman, Susan. / Comparison of LigAmp and an ASPCR assay for detection and quantification of K103N-containing HIV variants. In: AIDS Research and Human Retroviruses. 2008 ; Vol. 24, No. 4. pp. 595-605.
@article{7b513b22a8bb46d9a07ceaff737334b4,
title = "Comparison of LigAmp and an ASPCR assay for detection and quantification of K103N-containing HIV variants",
abstract = "We compared the ability of the LigAmp assay and an ASPCR assay to detect and quantify K103N-containing HIV variants in samples from 63 women who received single-dose nevirapine in a clinical trial. Samples were first analyzed with the ViroSeq HIV Genotyping system, and ViroSeq PCR products were used as templates for the LigAmp and ASPCR assays. A cutoff of 0.5{\%} K103N for detection of K103N was used for both assays. Results for the percentage K103N were similar for the two assays (R2 = 0.92). Forty-six samples (73.0{\%}) were positive for K103N by both assays and 13 samples (20.6{\%}) were negative by both assays. Four samples (6.3{\%}) were positive by ASPCR only. No samples were positive by LigAmp only. Eight discordant samples were analyzed in more detail. Sequence polymorphisms near oligonucleotide binding sites provided a possible explanation for the discordance in four of eight samples. The percentage K103N was also determined by analyzing 40 HIV clones from each of these eight samples, using a combined amplification/sequencing method (AmpliSeq). The percentage K103N determined by clonal analysis was consistent with the LigAmp result for five of eight samples, and was consistent with the ASPCR result for three of eight samples. Among 320 clones analyzed, we identified eight different codons at position 103 (mean = 3.8 codons/sample), which encoded six different amino acids, illustrating the extensive genetic diversity in HIV. Further studies are needed to compare performance of assays for detection and quantification of HIV drug resistance mutations in clinical samples.",
author = "Church, {Jessica D.} and Towler, {William I.} and Hoover, {Donald R.} and Hudelson, {Sarah E.} and Newton Kumwenda and Taha, {Taha E} and James Eshleman and Susan Eshleman",
year = "2008",
month = "4",
day = "1",
doi = "10.1089/aid.2007.0224",
language = "English (US)",
volume = "24",
pages = "595--605",
journal = "AIDS Research and Human Retroviruses",
issn = "0889-2229",
publisher = "Mary Ann Liebert Inc.",
number = "4",

}

TY - JOUR

T1 - Comparison of LigAmp and an ASPCR assay for detection and quantification of K103N-containing HIV variants

AU - Church, Jessica D.

AU - Towler, William I.

AU - Hoover, Donald R.

AU - Hudelson, Sarah E.

AU - Kumwenda, Newton

AU - Taha, Taha E

AU - Eshleman, James

AU - Eshleman, Susan

PY - 2008/4/1

Y1 - 2008/4/1

N2 - We compared the ability of the LigAmp assay and an ASPCR assay to detect and quantify K103N-containing HIV variants in samples from 63 women who received single-dose nevirapine in a clinical trial. Samples were first analyzed with the ViroSeq HIV Genotyping system, and ViroSeq PCR products were used as templates for the LigAmp and ASPCR assays. A cutoff of 0.5% K103N for detection of K103N was used for both assays. Results for the percentage K103N were similar for the two assays (R2 = 0.92). Forty-six samples (73.0%) were positive for K103N by both assays and 13 samples (20.6%) were negative by both assays. Four samples (6.3%) were positive by ASPCR only. No samples were positive by LigAmp only. Eight discordant samples were analyzed in more detail. Sequence polymorphisms near oligonucleotide binding sites provided a possible explanation for the discordance in four of eight samples. The percentage K103N was also determined by analyzing 40 HIV clones from each of these eight samples, using a combined amplification/sequencing method (AmpliSeq). The percentage K103N determined by clonal analysis was consistent with the LigAmp result for five of eight samples, and was consistent with the ASPCR result for three of eight samples. Among 320 clones analyzed, we identified eight different codons at position 103 (mean = 3.8 codons/sample), which encoded six different amino acids, illustrating the extensive genetic diversity in HIV. Further studies are needed to compare performance of assays for detection and quantification of HIV drug resistance mutations in clinical samples.

AB - We compared the ability of the LigAmp assay and an ASPCR assay to detect and quantify K103N-containing HIV variants in samples from 63 women who received single-dose nevirapine in a clinical trial. Samples were first analyzed with the ViroSeq HIV Genotyping system, and ViroSeq PCR products were used as templates for the LigAmp and ASPCR assays. A cutoff of 0.5% K103N for detection of K103N was used for both assays. Results for the percentage K103N were similar for the two assays (R2 = 0.92). Forty-six samples (73.0%) were positive for K103N by both assays and 13 samples (20.6%) were negative by both assays. Four samples (6.3%) were positive by ASPCR only. No samples were positive by LigAmp only. Eight discordant samples were analyzed in more detail. Sequence polymorphisms near oligonucleotide binding sites provided a possible explanation for the discordance in four of eight samples. The percentage K103N was also determined by analyzing 40 HIV clones from each of these eight samples, using a combined amplification/sequencing method (AmpliSeq). The percentage K103N determined by clonal analysis was consistent with the LigAmp result for five of eight samples, and was consistent with the ASPCR result for three of eight samples. Among 320 clones analyzed, we identified eight different codons at position 103 (mean = 3.8 codons/sample), which encoded six different amino acids, illustrating the extensive genetic diversity in HIV. Further studies are needed to compare performance of assays for detection and quantification of HIV drug resistance mutations in clinical samples.

UR - http://www.scopus.com/inward/record.url?scp=42449083333&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=42449083333&partnerID=8YFLogxK

U2 - 10.1089/aid.2007.0224

DO - 10.1089/aid.2007.0224

M3 - Article

C2 - 18370589

AN - SCOPUS:42449083333

VL - 24

SP - 595

EP - 605

JO - AIDS Research and Human Retroviruses

JF - AIDS Research and Human Retroviruses

SN - 0889-2229

IS - 4

ER -