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Journal of Clinical Microbiology, November 2000, p. 3919-3925, Vol. 38, No. 11
0095-1137/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.
Genetic Diversity of Protease and Reverse
Transcriptase Sequences in Non-Subtype-B Human Immunodeficiency Virus
Type 1 Strains: Evidence of Many Minor Drug Resistance Mutations in
Treatment-Naive Patients
Laurence
Vergne,1
Martine
Peeters,1,*
Eitel
Mpoudi-Ngole,2
Anke
Bourgeois,2
Florian
Liegeois,1
Coumba
Toure-Kane,3
Souleymane
Mboup,3
Claire
Mulanga-Kabeya,1
Eric
Saman,4
Jacques
Jourdan,5
Jacques
Reynes,6 and
Eric
Delaporte1,6
Laboratoire Retrovirus, IRD,
Montpellier,1 CHU, Caremeau, 30029 Nimes
cedex 4,5 and CHU, Gui de Chauliac,
34295 Montpellier Cedex 5,6 France;
Projet PRESICA, BP 906, Yaounde,
Cameroon2; CHU, Le Dantec, BP 7325, Dakar, Senegal3; and Innogenetics, 9052 Zwijnaarde, Belgium4
Received 24 February 2000/Returned for modification 11 May
2000/Accepted 17 July 2000
 |
ABSTRACT |
Most human immunodeficiency virus (HIV) drug susceptibility studies
have involved subtype B strains. Little information on the impact of
viral diversity on natural susceptibility to antiretroviral drugs has
been reported. However, the prevalence of non-subtype-B (non-B) HIV
type 1 (HIV-1) strains continues to increase in industrialized countries, and antiretroviral treatments have recently become available
in certain developing countries where non-B subtypes predominate. We
sequenced the protease and reverse transcriptase (RT) genes of 142 HIV-1 isolates from antiretroviral-naive patients: 4 belonged to group
O and 138 belonged to group M (9 subtype A, 13 subtype B, 2 subtype C,
5 subtype D, 2 subtype F1, 9 subtype F2, 4 subtype G, 5 subtype J, 2 subtype K, 3 subtype CRF01-AE, 67 subtype CRF02-AG, and 17 unclassified
isolates). No major mutations associated with resistance to nucleoside
reverse transcriptase inhibitors (NRTIs) or protease inhibitors were
detected. Major mutations linked to resistance to non-NRTI agents were
detected in all group O isolates (A98G and Y181C) and in one subtype J virus (V108I). In contrast, many accessory mutations were found, especially in the protease gene. Only 5.6% of the 142 strains, all
belonging to subtype B or D, had no mutations in the protease gene.
Sixty percent had one mutation, 22.5% had two mutations, 9.8% had
three mutations, and 2.1% (all group O strains) had four mutations. In
order of decreasing frequency, the following mutations were identified
in the protease gene: M36I (86.6%), L10I/V (26%), L63P (12.6%),
K20M/R (11.2%), V77I (5.6%), A71V (2.8%), L33F (0.7%), and M46I
(0.7%). R211K, an accessory mutation associated with NRTI resistance,
was also observed in 43.6% of the samples. Phenotypic and clinical
studies are now required to determine whether multidrug-resistant viruses emerge more rapidly during antiretroviral therapy when minor
resistance-conferring mutations are present before treatment initiation.
| |
INTRODUCTION |
Human immunodeficiency virus (HIV)
replication is markedly inhibited by highly active antiretroviral drug
combinations. Drugs belonging to three different classes
nucleoside
analogue reverse transcriptase (RT) inhibitors (NRTIs), non-NRTIs
(NNRTIs), and protease inhibitors (PIs)are currently used in various
combinations to treat HIV-infected patients (3, 14, 20, 43).
Replication of drug-resistant HIV type 1 (HIV-1) during combination
therapy is considered a major cause of treatment failure (18,
43). Drug resistance arises from mutations in the genes that
encode the molecular targets for the drugs, i.e., the RT and protease pol gene products. This viral polymorphism is due to the
high rate of HIV-1 replication and the low fidelity of RT (19, 30, 45). The emergence of amino acid substitutions associated with resistance to RT and PIs has been extensively characterized
(18, 34; http://hivdb.stanford.edu/hiv/), and
these substitutions can be classified into major and accessory
(modifying) mutations. Major mutations lead to a severalfold decrease
in sensitivity to one or more antiretroviral drugs (18).
Accessory mutations may not result in a significant decrease in
sensitivity but are associated with an increase in viral fitness
(replication capacity) (14, 18). Thus, the appearance of a
major mutation in a genome already containing accessory mutations could
influence the speed with which highly resistant viruses are selected
during therapy.
Genetic characterization and phylogenetic analysis of HIV-1 isolates
from different geographic localities have revealed that HIV-1 can be
divided into at least three distinctive groups, designated M (major), N
(new or non-M, non-O), and O (outlier) (38). Group M
comprises most of the HIV-1 strains responsible for the AIDS pandemic
(15) and can be further subdivided into subtypes (subtypes A
to K) (8, 42). Recombination events among sequences of different genetic subtypes of HIV-1 group M have frequently been identified (32, 33). Some of these mosaic HIV-1 genomes are unique, but others play a major role in the AIDS pandemic and are named
circulating recombinant forms (CRFs) (8). They are designated, according to new nomenclature proposals (33a),
by an identifying number and letters indicating the source subtypes, e.g., CRF01-AE (initially env subtype E) and CRF02-AG
(AG-IBNG-like viruses) (9, 16).
Most HIV-1 isolates in North America and Europe belong to subtype B. Therefore, anti-HIV drug testing and characterization of drug
resistance mutations that confer resistance have been done in studies
with subtype B isolates, but subtype B isolates are the cause of only a
limited proportion of infections worldwide (15). The
efficacy of antiretroviral treatment can be influenced by the viral
subtype. Like HIV-2, HIV-1 group O viruses are naturally resistant to
NNRTIs (11, 28, 31). Within group M, some subtype F samples
are less susceptible to the tetrahydroimidazo [4,5,1-jk] [1,4]-benzodiazepin-2-(1H)-one and -thione (TIBO) derivative, an
NNRTI, and some subtype G strains have decreased susceptibility to PIs
(2, 12).
With the increasing frequency of non-subtype-B (non-B) isolates in
Europe (1, 4, 13) and the recent introduction of antiretroviral drugs in developing countries, there is a need to test
the efficacies of existing and new drugs against non-B strains and to
monitor resistance. Natural mutations that confer drug resistance have
been described in drug-naive patients infected with subtype B strains
before the drugs were first used in the relevant population (6, 7,
24, 26, 36), but the prevalence of such mutations has not been
routinely studied in untreated individuals infected with non-B strains
(10, 29, 39). In this study, we analyzed the protease and RT
sequences of isolates from 129 treatment-naive patients infected with
non-B HIV-1 strains for the presence of natural mutations linked to
resistance to antiretroviral drugs.
| |
MATERIALS AND METHODS |
Patients.
One hundred forty-two antiretroviral drug-naive,
HIV-1-infected patients were studied. Samples were collected between
1995 and 1999 from patients from four distinct sites: 62 from Cameroon, 38 from Senegal, 18 from the Democratic Republic of Congo (DRC; formerly Zaire), and 24 from a hospital in southern France. Among the
24 patients recruited in France, 9 were infected in Europe, 2 were
infected in Cambodia, and 13 were infected in Africa (Ivory Coast
[n = 3], Guinea Conakry [n = 1],
Central African Republic [CAR; n = 3], Burkina Faso
[n = 1], and Cameroon [n = 1]); the country of infection was unknown for the other 4 patients).
RNA and DNA extraction, cDNA synthesis, and PCR.
For samples
from DRC and Cameroon, proviral DNA was extracted from uncultured
peripheral blood mononuclear cells and from cultured peripheral blood
mononuclear cells, respectively, with the QIAamp Blood and Tissue kit
(QIAGEN, Courtaboeuf, France). For samples from Senegal and France,
viral RNA was isolated from plasma with the QIAamp Viral RNA Mini kit (QIAGEN).
Viral RNA was transcribed to cDNA by using Expand RT (Boehringer
Mannheim, Germany) with a reverse primer (primer IN3
[5'-TCTATBCCATCTAAAAATAGTACTTTCCTGATTCC-3'] in RT fragment
at position 4261) (44). Reverse transcription was carried
out in a final volume of 20 µl containing 1× Expand RT buffer, 10 mM
dithiothreitol, 1 mM each deoxynucleoside triphosphate, 40 pmol of
reverse primer, and 50 U of Expand RT at 42°C for 60 min and 92°C
for 2 min. A 2,200-bp fragment encompassing the protease and RT genes
was amplified from DNA or cDNA with the Expand Long Template PCR system
(Boehringer, Mannheim, Germany) by a seminested PCR method with outer
primers G25REV (5'-GCAAGAGTTTTGGCTGAAGCAATGAG-3'; at
position 1873) and IN3 and inner primers AV150
(5'-GTGGAAAGGAAGGACACCAAATGAAAG-3'; at position 2042) and
IN3 (44). PCRs were carried out in final volumes of 50 µl
(first round) and 100 µl (second round). The reaction mixture
consisted of 1× PCR buffer with 0.75 mM MgCl2, 0.2 mM each
deoxynucleoside triphosphate, 20 pmol of each primer, and 2.5 U of
Expand Long Template enzyme mixture. The PCR conditions were 92°C for
5 min, followed by 39 cycles at 92°C for 20 s, 50°C for
30 s, and 72°C for 2 min, with a final extension step at 72°C for 10 min.
Sequencing reactions.
The amplified fragments were purified
with the QIAquick gel extraction kit (QIAGEN) and were directly
sequenced with the ABIPRISM BigDye Terminator Cycle Sequencing
Ready Reaction Kit with AmpliTaq DNA polymerase (FS; Perkin-Elmer,
Roissy, France) on an automated sequencer (373A stretch; Applied Biosystems).
Phylogenetic and sequence analyses.
The genetic subtypes
were determined by phylogenetic tree analysis. The new nucleotide
sequences and the sequences of the protease and RT genes from reference
strains representing the different genetic subtypes were aligned with
the CLUSTAL W program, bearing in mind the protein sequences
(40). Phylogenetic trees constructed by the neighbor-joining
method and the reliability of the branching orders obtained by the
bootstrap approach were implemented with the CLUSTAL W program. Genetic
distances were calculated by the two-parameter method of Kimura
(22). To determine whether the viruses were recombinants in
the sequenced region, several additional analyses were performed.
Diversity plots, obtained with the DIVERT program (available online
[http://193.50.234.246/~beudoin/anrs/Diversity.html]) were used to
determine the percent diversity between selected pairs of sequences by
moving a window of 300 bp along the genome alignment in 20-bp
increments. The divergence values for each pairwise comparison were
plotted at the midpoint of the 300-bp segment. Simplot, version 2.5, software (http://www.med.jhu.edu/deptmed/sray/download) was used to
calculate bootstrap plots by bootscanning the neighbor-joining trees
with SEQBOOT, DNADIST (by the two-parameter method of Kimura [22] and with a transition/transversion ratio of 2.0),
NEIGHBOR, and CONSENSUS from the Phylip package for a 300-bp window
moving along the alignment in increments of 20 bp. We evaluated 100 replicates for each phylogenetic analysis. The bootstrap values for the
sequences studied were plotted at the midpoint of each window.
The amino acid sequences of the protease and RT genes, deduced from the
nucleic acid sequences, were compared to a subtype
B consensus sequence
from the Stanford HIV RT and Protease Sequence
database
(
37;
http://hivdb.stanford.edu/hiv/) and
analyzed for
mutations associated with reduced sensitivity to
antiretroviral
drugs. A consensus
pol (protease and RT)
sequence was calculated
for each subtype with VESPA (Viral
Epidemiologic Signature Pattern
Analysis) software (
23), and
signatures specific to each subtype
were deduced relative to subtype
B.
Nucleotide sequence accession numbers.
The protease and RT
sequences are available in GenBank (EMBL) with the following accession
numbers: AJ286930 to AJ286979, AJ286133 to AJ286135, AJ286137 to
AJ286140, AJ286142, AJ286143, AJ249236, AJ249237, and AJ249239 for
sequences from Cameroon; AJ286980 to AJ287014, AJ286136, AJ286141, and
AJ251057 for sequences from Senegal; AJ287015 to AJ287031 and AJ249235
for sequences from DRC; and AJ287032 to AJ287054 and AJ249238 for
sequences from France.
| |
RESULTS |
Phylogenetic analysis of the protease and RT sequences.
Table
1 summarizes the genetic subtypes in the
pol region, and Fig. 1 shows
the phylogenetic trees for the HIV-1 group M sequences from Senegal,
Cameroon, DRC, and France. Phylogenetic tree analysis of all the
isolates together showed that no laboratory contamination had occurred
(data not shown). Four of the 142 isolates tested belonged to group O
(1 isolate from Cameroon and 3 isolates from Senegal). Among the
remaining 138 group M isolates, the overall subtype distribution was as
follows: 9 subtype A, 13 subtype B, 2 subtype C, 5 subtype D, 2 subtype
F1, 9 subtype F2, 4 subtype G, 5 subtype J, 2 subtype K, 3 subtype
CRF01-AE, and 67 subtype CRF02-AG. The subtypes of 17 HIV-1 strains
could not be clearly determined, as they did not cluster with any of
the known subtypes.
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FIG. 1.
Unrooted phylogenetic trees of protease and RT
nucleotide sequences (1,600 bp) from 138 group M isolates (in gray) and
from reference strains (in black) representing the different subtypes.
The trees were generated as described in Materials and Methods. The
reference sequences (8) used in the trees were as follows:
A.U455, A.Q2317, A.92UG037.1, B.OYI, B.HXB2, B.JRFL, B.RF, D.NDK,
D.ELI, D.84ZR085, D.94UG114.1, C.ETH2220, C.92BR025.8, F1.93BR020.1,
F1.FIN9363, G.SE6165, G.92NG083, G(A).92NG003, G.HH8793.1.1, H.VI991,
H.VI997, H.90CF056.1, J.SE9173.3, J.SE9280.9, CRF01-AE.CM240,
CRF01-AE.93TH253.3, CRF01-AE.90CF402.1, CRF02-AG.IBNG, and
CRF02-AG.DJ263. The reference sequences for the K and F2 subtypes
(41, 42) were used either as references or as samples
(K.96CAM.MP535, K.97ZR.EQTB11, F2.95CM.MP255, F2.95CM.MP257). For
sequences that did not cluster with high bootstrap values with any of
the known subtypes or CRFs, the results of complementary analyses
indicating their recombinant nature are shown.
a, the isolates designated A/G and G/A/G are
intersubtype A/G recombinant viruses in which the breakpoints between A
and G are different from those for the prototype CRF02-AG strain. The
numbers at the branch points indicate bootstrap values as
percentages.
|
|
More detailed analysis of the 17 unclassified strains by bootscanning
and with the Divert program showed that 15 were recombinants
in the
pol region studied. Six isolates from DRC were A/G
recombinants
(but different from the CRF02-AG strains) and formed a
distinct
cluster in the phylogenetic tree; another two isolates from
DRC
were G/A/G recombinants. Two isolates (one isolate from Senegal
and
one isolate from a patient from Burkina Faso attending a hospital
in
France) were G/K recombinants. One Cameroonian isolate was
a D/G/D
recombinant in the studied region. Four isolates (one
isolate from
Cameroon, one isolate from Senegal, and two isolates
from DRC) had a
similar recombinant profile, with an unclassified
protease and an
unclassified 5' end of the RT gene and with the
3' end of the RT being
subtype K. Finally, two samples from DRC
did not cluster with any of
the known subtypes over the entire
protease and RT
regions.
Isolates of almost all known subtypes and CRFs were represented in our
study. CRF02-AG was predominant, representing most
of the viruses
circulating in Senegal and Cameroon and non-B isolates
in France. These
strains are subtype G in the protease gene and
at the 5' end of the RT
gene and subtype A at the 3' end of the
RT
gene.
Protease sequence variability in isolates from untreated non-B
HIV-1-infected patients.
The amino acid sequence of each strain
was compared to the subtype B consensus amino acid sequence (Stanford
HIV protease sequence database) for mutations associated with
resistance to protease inhibitors (37;
http://hivdb.stanford.edu/hiv/). No major mutations (D30N, G48V, I50V,
V82A/T/F, I84V, or L90M) were seen in any of the subtype B or non-B
strains from our sample. In contrast, many minor or accessory mutations
were found at the following positions, in order of decreasing
frequency: M36I (n = 123 [86.6%]), L10I/V
(n = 37 [26%]), L63P (n = 18 [12.6%]), K20M/R (n = 16 [11.2%]), V77I
(n = 8 [5.6%]), A71V (n = 4 [2.8%]), L33F (n = 1 [0.7%]), and M46I
(n = 1 [0.7%]). No accessory mutations were seen at
positions F53L, G73S, and N88D. Table 2
summarizes the frequencies of the different mutations according to
subtype. No mutations specific for a given subtype were noted, but the M36I mutation was present in 123 of the 129 non-B strains but was not
present in any of the subtype B strains. In general, subtype B strains
(n = 13) bore few mutations (only L63P [n = 6] and V77I [n = 3]). Group O strains bore the
resistance-conferring minor mutations L10I/V, M36I, L63P (n = 3), and A71V, with the last mutation being specific for group O
viruses.
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TABLE 2.
Amino acid substitutions in the HIV-1 protease sequences
of isolates from 142 treatment-naive patients at key positions
associated with resistance to available drugs
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|
Considering all the minor mutations related to resistance, only 8 (5.6%) of the 142 strains had no mutations, and they were
all of
subtype B or D. Eighty-five strains (60%) had one mutation,
32 (22.5%) had two mutations, 14 (9.8%) had three mutations, and
3 (2.1%) (all group O) had four
mutations.
Amino acid changes associated with resistance to protease inhibitors
(ABT378 and DMP450) currently being tested in clinical
trials were also
identified: G16E (
n = 27) and D60E (
n = 8) (
17).
In addition, at some key positions (i.e., positions known for amino
acid mutations associated with drug resistance), amino
acid
substitutions different from those linked to PI resistance
were
observed. Eighty-five strains (59.8%) had an amino acid different
from
lysine (K) at position 20: K20I (
n = 79), K20C
(
n = 4), and
K20V (
n = 2). The K20I
substitution was found in 63 of the 67
CRF02-AG strains, in all subtype
G strains, in all A/G and G/A/G
recombinant strains from DRC, and in
the G/K recombinants. Thirty-two
strains showed a polymorphism other
than L63P. For all the subtype
G strains, the G/A/G recombinants, and
the three CRF02-AG viruses,
the valine (V) at position 82 was replaced
by an isoleucine
(I).
Figure
2 shows the alignment of the amino
acid consensus sequences of the protease gene for each subtype compared
with a subtype
B consensus sequence from the database. The subtype B
consensus
sequence from the isolates from the 13 HIV-1 subtype
B-infected
patients in our study corresponds to the subtype B consensus
sequence
from the database. Overall, the three functional sites in
which
major mutations are located (except L90M) were highly conserved
among all the subtypes. Leucine (L) was replaced by a methionine
(M) at
position 89 of all group M subtype isolates (except isolates
of
subtypes B and D) and by an isoleucine (I) in group O isolates.
In the
nonfunctional sites of protease, more amino acid substitutions
were
seen in the group O sequences than in the group M sequences.
Other
mutations specific to certain subtypes were seen in other
regions of
the protease gene by using the VESPA program (80% threshold):
13V for
subtypes A, G, and CRF02-AG; 15V for subtype K; 35D for
subtypes A,
CRF01-AE, and F; 41K for all the subtypes except J;
61F and 67C for
subtype J; 69K for all the subtypes except F and
K; 89M for all the
subtypes except D and K; and 93L for subtype
C.
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FIG. 2.
Amino acid alignment of protease consensus sequences for
each subtype of group M and for group O. The sequences were aligned
against a subtype B consensus sequence from the database; dots indicate
homology. The amino acid positions associated with drug resistance are
depicted in boldface italics; the major mutations (D30N, G48V, I50V,
V82A/F/T, I84V, L90M) are marked at the top of the consensus sequence
( ), as are the minor mutations (L10I/V, K20M/R, L24I, V32I, L33F,
M36I, M46I/L, I47V, I54L/V, L63P, A71T/V, G73S, V77I, N88D) ( ). The
functional domains of the protease (*, active site [amino acids 22 to 34]; **, flap region [amino acids 47 to 56]; ***,
substrate binding site [amino acids 78 to 88]) are shown under the
consensus sequence.
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|
Sequence variability in RT region of isolates from untreated non-B
HIV-1-infected patients.
The mutations leading to resistance to
NRTIs and NNRTIs are well defined and differ between the two classes of
RT inhibitors (18, 34).
None of the strains had major mutations associated with NRTI
resistance. Only one patient, a European infected with a subtype
B
isolate from a zidovudine (AZT)-treated partner, had the accessory
mutation M41L, along with T215D, which represents a transition
to the
T215Y/F major mutation associated with resistance to AZT
(
37;
http://hivdb.stanford.edu/hiv/). Accessory
mutations were
also observed in the RT gene: 62 (43.6%) of the
strains, representing
all subtypes, had the R211K mutation, and 5 strains had the G333E
mutation.
In some strains, polymorphisms were observed at key positions but were
not associated with resistance, namely, L210Y (
n =
4)
and L210Q (
n = 4; all group O), R211S (
n = 10), and R211N (
n = 2).
Major mutations associated with NNRTI resistance were seen: V108I in
one subtype J isolate and A98G and Y181C in the four
group O isolates
(
18;
http://hivdb.stanford.edu/hiv/). Other
amino
acid changes not associated with drug resistance were observed
at key
positions in a few strains, namely, A98S (
n = 8) and
K101R
(
n = 1).
Amino acid changes associated with resistance to NNRTIs being tested in
clinical trials were also identified, namely, V106I
(
n = 3) (HBY097), V179D (
n = 1) (trovirdine, QM96521,
UC-10, ADAMII,
L-697,661, and TIBO R82913), V179E (
n = 6) (L-697,661), and V189I
(
n = 1) (HBY097)
(
17).
| |
DISCUSSION |
We analyzed the protease and RT gene-coding regions of 142 HIV-1
isolates from treatment-naive patients. Most strains were isolated in
Africa, where access to antiretroviral drugs is very limited. More than
90% of the strains were non-B, and isolates of all genetic subtypes
with the exception of subtype H and two CRFs were represented. Isolates
of subtype CRF02-AG, which is predominant in west and west-central
Africa (27), represented 47% of the isolates.
Overall, the protease gene region was less conserved than the RT gene
region. No major mutations conferring resistance to PIs were seen, but
more than 34% of the strains had two or more minor mutations
associated with PI resistance; only 5.6% of strains (mainly subtype B
strains) had no minor mutations. M36I, the predominant minor mutation,
was observed in 87% of the strains overall and in 95% of the non-B
strains. Amino acid substitutions associated with PI resistance have
been reported as natural variants in treatment-naive patients (5,
6, 10, 24, 25, 29, 35, 36, 39), but the prevalence of
substitutions determined from our data are significantly higher than
those from previous studies with subtype B isolates (10, 24, 37,
39). Several mutations not classically associated with resistance
were observed, and their biological consequences remain to be studied.
Accessory mutations are not always associated with a decrease in in
vitro susceptibility, in contrast to major mutations (18,
43). For example, the protease sequences of subtype C strains
isolated in Zimbabwe had several accessory mutations, but in vitro
susceptibility tests with a subset of these samples showed no decrease
in sensitivity (35). Minor mutations can compensate for the
reduced fitness of resistant mutants, and in contrast to major
mutations, which are different for each protease inhibitor,
compensatory mutations are similar for many PIs (14, 18,
43). The efficacy of a switch from one PI to another might
therefore be compromised when the virus has had the opportunity to
develop compensatory mutations. One consequence of preexisting
accessory mutations might be the faster emergence of viruses resistant
to PIs. Preliminary data suggest that prior M36I and L10I/V mutations
are associated with a more rapid fall in sensitivity during treatment
(C. F. Perno, A. D'Arminio-Monforte, A. Cozzi-Lepri, C. Balotta,
F. Forbici, A. Bertoli, P. Pezzotti, G. Facchi, L. Monno, G. Angarano,
P. Bottura, V. Vullo, A. Cargnel, M. Capobianchi, G. Ippolito, and M. Moroni, 7th Conference on Retroviruses and Opportunistic Infections, 2000).
No major mutations conferring resistance to NRTIs were seen, but two
accessory mutationsR211K (43.6%) and G333E (3.5%)were found.
These mutations facilitate dual resistance to AZT and lamivudine (3TC)
in association with M184V and other AZT resistance mutations (17,
21). In contrast to NRTIs, only major mutations associated with
resistance to NNRTIs were seen. A pol subtype J isolate from one patient originally from CAR had a mutation (V108I) associated with
resistance to nevirapine and efavirenz. In keeping with previous reports, major mutations to NNRTIs were present in all four group O isolates.
In conclusion, the prevalence of major mutations associated with
resistance to NRTIs, NNRTIs, and PIs is very low among non-B group M
HIV-1 isolates of African origin, whereas many minor or accessory
mutations related to resistance to NRTIs and PIs are present as natural
variants. Phenotypic and clinical studies are necessary to determine
whether, when antiretroviral therapy is started, these viruses generate
multidrug-resistant strains more rapidly.
| |
ACKNOWLEDGMENTS |
This study was cosponsored by grants from the Agence Nationale de
Recherche contre le SIDA (ANRS; project SIDAK), the European Union
(INCO-DC; grant IC18CT97-0216), and SIDACTION.
| |
FOOTNOTES |
*
Corresponding author. Mailing address: Laboratoire
Retrovirus, IRD, 911 Avenue Agropolis, BP 5045, 34032 Montpellier Cedex 1, France. Phone: 33-4 67 41 62 97. Fax: 33-4 67 61 94 50. E-mail: martine.peeters{at}mpl.ird.fr.
| |
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Journal of Clinical Microbiology, November 2000, p. 3919-3925, Vol. 38, No. 11
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Abstract
Introduction
