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L Zekeng

Publications and source records attributed to L Zekeng.

At least 55 records · Page 3Linked to original sources
PubMed1997

Envelope sequence variability and serologic characterization of HIV type 1 group O isolates from equatorial guinea.

Four sera from Equatorial Guinea (EG) suspected to contain antibody against HIV-1 group O-related viruses were identified on the basis of unusual and differential serologic reactivity in selected commercial assays and Western blot. Degenerate primers, designed from HIV-1 group O published sequences, were used to PCR amplify envelope (env) gene sequences from the suspect EG sera. A complete envelope gene sequence from each serum was determined from the overlapping env gene fragments. Analysis (PHYLIP package of programs) of Env amino acid sequences (translated from nucleotide sequences) indicated that the amino acid sequences obtained from EG sera clustered more closely with HIV Env sequences of group O compared to group M. The amino acid sequences at the octameric tip of the V3 loop were either RIGPLAWY (one isolate), RIGPMAWY (two isolates), or GLGPLAVY (one isolate). The V3 tip tetrameric sequence GPLA is represented only once in the 1995 HIV (Los Alamos) database, but was present in two of our group O-related EG samples. The gp41 immunodominant regions (IDR) protein sequences were identical for sequences from three of the sera, RLLALETLIQNQQLLNLWGCKGR(K)L(I)VCYTSVK(T)W, whereas sequence from the fourth serum contained three changes as noted in parentheses. IDR sequences derived from EG sera were unique compared to those reported for other HIV-1 group O isolate ANT70, VAU, or MVP5180. Antibody in each EG serum directed against the IDR could be detected using synthetic peptides comprising sequences from the ANT70 or MVP5180 IDRs, but were most reactive against the sequences derived from the samples themselves. Little or no serologic reactivity was detected when EG sera were reacted against peptides comprising the IDR of HIV-1 group M (subtype B consensus) or HIV-2 (consensus).

Amino Acid Sequence↗
PubMed1997

Evaluation of a quantitative double ELISA strategy for confirmation and differentiation of HIV infection.

The current HIV pandemic is complicated by the spread of distinct types and subtypes of HIV. The currently used conventional diagnostic tests have shown limitations in the detection of antibodies against all HIV-1 subtypes, as demonstrated by recent identification of HIV-1 subtype O. To evaluate quantitatively the diagnostic potential of a double ELISA strategy for the detection and partial differentiation of HIV-1, HIV-1 subtype O and HIV-2 infections blood samples were examined at five different test centers: Blantyre, Malawi; Abidjan and Daloa, Ivory Coast; Yaoundé, Cameroon; Munich, Germany. All tests results, including ELISA extinction values and Western blot profiles, were forwarded to Munich for final interpretation. An indirect anti-HIV-1/2 ELISA and a competitive anti-HIV-1 ELISA were used in combination for the initial screening of blood specimens. All anti-HIV positive and anti-HIV negative samples were subjected to immunoblot analysis. Independent of the diversity of the extinction profiles, and of the test manufacturer, the quantitative evaluation of the ELISA extinction values could define two extinction areas with a 100% predictive value for HIV-1 seropositivity and HIV seronegativity; extinction values > 2 by the indirect ELISA and < 0.2 by the competitive ELISA for an anti-HIV-1 subtype A to I positive result; extinction values < 0.2 by the indirect ELISA and > 1.0 by the competitive ELISA for an anti-HIV negative result. Additionally, the quantitative evaluation of the extinction profile provides partial information on the HIV-1 subtype as far as the distinction in group M and group O is concerned. In conclusion, the quantitative evaluation of this double ELISA strategy can reduce the number of blood specimens that require additional confirmatory testing in developing countries and can be superior to the immunoblot method during early seroconversion.

Antigens, Viral↗
PubMed1997

Diversity of the immunodominant epitope of gp41 of HIV-1 subtype O and its validity for antibody detection.

The immunodominant regions of the gp41 from 13 HIV-1 subtype O strains from Cameroon, 11 from France and one from Germany were sequenced. The amino acid sequences were compared to those of the 3 published HIV-1 subtype O isolates, ANT70, MVP-5180 and VAU. All HIV-1 subtype O isolates had a very conserved amino acid sequence in this region and showed a subtype O specific structure. Within the cysteine loop there was a positive charge of two basic amino acids, arginine and lysine. Only two strains (CM.6778 and CM.8161) showed an acidic amino acid in this loop. None of the isolates showed the same amino acid sequence in this immunodominant region. A 25 residue peptide from the immunodominant domain of gp41 of the MVP-5180 strain was synthesized, cycled to form the cysteine-loop and coated to microtiter plates. Antibody binding was detected by indirect ELISA using an enzyme labeled anti-human IgG. Out of 111 anti-HIV-1 positive specimens, collected mainly from Cameroonian HIV infected patients, only 10 were not reactive in this assay. The 42 anti-HIV-1 subtype O positive specimens gave all a reaction above cut off. Despite the diversity found in the amino acid sequences within the 25 isolates a peptide-based indirect ELISA representing the immunodominant epitope of the strain MVP-5180 successfully detected all the anti-HIV-O sera so far tested, pointing to the importance of adding such a peptide for correct identification of HIV-1 subtype O infected patients, while some assays without HIV-O specific antigens partially fail to detect all anti-HIV-O specimens.

Amino Acid Sequence↗
PubMed1997

Reactivity of a new HIV-1 group O third generation A-HIV-1/-2 assay with an unusual HIV-1 seroconversion panel and HIV-1 group O/group M subtyped samples.

It was shown previously that about 97% of the anti-HIV-1 group O strain-positive samples were detected by crossreaction with native HIV-1 gp160 (Van Binsbergen et al., Evaluation of a new third generation anti-HIV-1/anti-HIV-2 assay with increased sensitivity for HIV-1 group O, J. Virol. Methods 60 (1996) 131-137). Fourteen out of 17 new anti-HIV-1 group O positive samples, selected with the Enzygnost HIV-1/2 plus assay, were already reactive when tested with HIV-1 gp160. When tested by the Vironostika HIV Uni-Form II plus O microELISA all 17 samples were reactive, demonstrating the necessity to implement an HIV-1 group O-specific antigen in the assay. On the other hand, it was surprisingly found that 40 out of 43 (93%) of anti-HIV-1 group M-positive samples, belonging to strain A, B, C, D, E or F, were detected by crossreaction with the HIV-1 group O (strain ANT70) synthetic peptide incorporated in the Vironostika HIV Uni-Form II plus O. Only HIV-1 subtype D-positive samples did not react with this peptide, presumably because of the presence of a histidine residue in the immunodominant region of HIV-1 subtype D gp41. Both crossreactions make the Vironostika HIV Uni-Form II plus O microELISA also sensitive for anti-HIV-1-positive samples originating from different geographical regions and resulting from different HIV-1 subtype infections. With an unusual seroconversion panel in which p24 Ag was present persistently, many anti-HIV-1/-2 assays produce alternating positive/negative results in anti-HIV antibody-positive bleeds. It was shown that the use of viral p24 and gp160 in a direct sandwich, allowing detection of anti-HIV IgG and IgM, explains the identification of all anti-HIV-positive bleeds by the Vironostika HIV Uni-Form II plus O. The high sensitivity of the plus O assay was confirmed with clinical samples of a so-called anti-HIV-1 low titer panel. The specificity of the Vironostika HIV Uni-Form II plus O determined in five blood transfusion centers, based on 135070 tests, was 99.97%.

AIDS Serodiagnosis↗
PubMed1997

Prevalence and phylogenetic analysis of HTLV-I isolates in Cameroon, including those of the Baka Pygmy.

Our previous analysis of an HTLV-I isolate (CMR229) from a Cameroonian Pygmy demonstrated that the isolate is distinct from typical HTLV-Is of the "Central African group," which has a close similarity to HTLV-I-related simian viruses (STLV-I) in Africa. In this study, we analyzed six new HTLV-Is from Cameroon consisting of three isolates from the Pygmy and three from the Bantu to examine further the genetic features of HTLV-I in Cameroon, especially in the Pygmy. A phylogenetic tree based on the long terminal repeats (LTR) region showed that all the new HTLV-Is belong to the Central African group. On the other hand, an env-based analysis of CMR229 confirmed the previous finding derived from LTR-based analysis that CMR229 has a similarity to African STLV-Is, but is distinct from the typical Central African group of HTLV-I. This suggests that multiple interspecies transmissions from non-human primates to humans have occurred in Central Africa, resulting in the presence of two distinct HTLV-I strains in this area. In addition, it seems likely that the Pygmy harbors the heterogeneous HTLV-I strains from which the main HTLV-I population spread into the Bantu.

Base Sequence↗
PubMed1997

Detection of genetically diverse human immunodeficiency virus type 1 group M and O isolates by PCR.

A panel of 136 genetically diverse group M and 5 group O adult isolates from outside the United States and Europe were evaluated by PCR with the Roche AMPLICOR HIV-1 test, a modified version of the AMPLICOR HIV-1 test, and a new primer pair/probe system. Detection of some of these isolates was less efficient with the AMPLICOR HIV-1 test; however, the assay was significantly improved by reducing the sample input and lowering the annealing temperature. The new primer pair/probe set detected 140 of 141 isolates, including the 5 group O isolates that were not detected with either of the AMPLICOR HIV-1 test formats.

Adult↗
PubMed1997

Molecular analyses of HIV-1 group O and HIV-2 variants from Africa.

Genetic variation among HIV isolates creates challenges for their detection by serologic and genetic techniques. To characterize the sequence variation and its correlation to serologic diversity of HIV-1 Group O and HIV-2 isolates, samples were identified by differential reactivity in selected commercial and research assays. Analysis of sera from Equatorial Guinea (EG) led to identification of 4 HIV-1 Group O variants. Viral RNA, extracted from these samples was used to PCR amplify overlapping sequences of the entire envelope gene using multiple primer pairs. Sequence analysis indicated that the V3 loop nucleotide and protein sequences aligned more closely with HIVANT70 compared to other Group O sequences. The amino acid sequences at the octameric tip of the V3 loop were RIGPLAWY, RIGPMAWY, or GLGPLAVY. The tetrameric tip GPLA is represented only once in the published 1994 HIV database (Los Alamos) but was present in 2 of 4 of EG samples. The immuno-dominant region (IDR) sequences derived from EG sera were unique in that none of the sequences were completely homologous to other HIV-1 group O variants. Further, the HIV-1 group O sequence variation could be correlated with differential serologic reactivity using IDR peptides. Compared to HIV-1, the sequence information on HIV-2 isolates is relatively limited, though the HIV-2 isolates also show genetic variation similar to HIV-1. To further establish a correlation between the genetic diversity and serologic detection of HIV-2, plasma samples from Western Africa were evaluated. Eight samples were selected based on weak serologic reactivity to env proteins. PCR amplification and sequence analysis of the gag, env V3 loop, and env IDR regions indicated that the samples could be classified as subtypes A (4 samples), B (3 samples) and D (1 sample). Across the subtypes, there was conservation in the IDR region of the sequence WGCAFRQVCHT. This region is absolutely conserved among the majority of currently known HIV-2 and related SIV viruses (1994 HIV database). One subtype B sample had a unique sequence immediately adjacent to the IDR, however, this did not change the serologic detection using a HIV-2 IDR specific monoclonal antibody.

Acquired Immunodeficiency Syndrome↗
PubMed1996

HIV-1 subtype O: epidemiology, pathogenesis, diagnosis, and perspectives of the evolution of HIV.

HIV-1 subtype O is a new HIV variant originating in the West-Central African region, with highest prevalences in countries such as Cameroon, Equatorial Guinea and Gabon. Detection of antibodies to HIV-1 subtype O can pose problems in unmodified ELISA tests, and confirmation of anti-HIV-1 subtype O in immunoblot may give false negative results in some specimens. Nucleic acid-based assays designed for HIV-1 detection do not amplify or detect sequences from HIV-1 subtype O. In their env sequences, HIV-1 subtype O strains show a higher heterogeneity than the classical HIV-1 subtypes, leading to the conclusion that HIV-1 subtype O has been introduced into the human population only recently. Further, unidentified subtypes are also likely to exist.

Acquired Immunodeficiency Syndrome↗
PubMed1996

Evaluation of a new third generation anti-HIV-1/anti-HIV-2 assay with increased sensitivity for HIV-1 group O.

Although the HIV-1 group O virus found in two persons of Cameroonian origin has been described in 1990 (De Leys et al., 1990), sera from group O infected individuals became available only recently. Several studies showed that some of the anti-HIV-1/HIV-2 screening tests failed to detect all of these samples (Loussert-Ajaka et al., 1994; Simon et al., 1994; Schable et al., 1994; Gürtler et al., 1995). In the current version of an anti-HIV-1/anti-HIV-2 screening assay, namely the Vironostika HIV Uni-Form II, an HIV-O specific peptide was introduced in order to improve HIV-1 group O reactivity. The peptide was derived from the immunodominant region of HIV-1 group O gp41 strain ANT70. All 30 anti-HIV-1 group O sera were detected by the so-called plus O assay, while 29 samples of this panel were positive the current assay. The sensitivity of the plus O assay for anti-HIV-1 and anti-HIV-2 positive samples is identical to that of the reference test without HIV-1 group O peptide. The clinical specificity of the HIV Uni-Form II plus O assay is improved to > or = 99.92% by an adjustment of the coat concentration of HIV-1 p24 (to avoid false positive p24 only reactions) without affecting sensitivity of the assay. The specific reaction of an HIV-1 group O specific rabbit serum for quality control purposes is presented.

AIDS Serodiagnosis↗
PubMed1996

[Cytologic aspects of cervical smears in optic microscopy in HIV seropositive women in Yaounde-Cameroon (Central Africa)].

Cytologic aspects of cervical smears in HIV seropositive women in Yaounde-Cameroon (Central Africa). The aim of this study was to present the cytologic aspects of cervical smears performed on HIV seropositive Cameroonian women and analysed by light microscope. Seropositive women (case group) and seronegative women (control group) had cervical smears which were stained by the Papanicolaou Method and analysed by light microscope. For the 65 seropositive women, there were: 62 inflammatory smears (95.5%), 2 normal (3%) and 1 low grade squamous intra-epithelial lesion (1.5%). The 50 seronegative women had: 35 inflammatory smears (70%), 13 normal (26%) and 2 low grade squamous intra-epithelial lesion (4%). In both groups, inflammatory smears were predominant. They were more frequent in seropositive women. There was no significant difference between the percentage of squamous intra-epithelial lesions in seropositive women (1.5%) and seronegative women (4%). We were unable to detect, in Cameroonian seropositive women, any specific lesions on cervical smears predictive of HIV infection without serology.

Adolescent↗
PubMed1995

Reactivity of five anti-HIV-1 subtype O specimens with six different anti-HIV screening ELISAs and three immunoblots.

Five anti-subtype O specimens were tested by anti-HIV-1/2 screening and confirmatory assays. They can be divided into three specimens, reactive with all ELISAs, independent of the nature of the antigen (recombinant proteins or peptides) and test configuration (indirect ELISA or double antigen/sandwich ELISA). One specimen was not detected by one peptide based ELISA. One specimen was only recognized by two ELISAs and should be considered as a marker sample for the weakness of currently used ELISAs with anti-subtype O. Three different immunoblot assays available commercially detected two of the specimens with a major binding of gp160 and other viral bands, especially the integrase and reverse transcriptase. Another two specimens lacked reactivity with glycoproteins almost completely, but showed some staining with the enzymes of HIV, and would most probably be interpreted as indeterminate. The fifth specimen, which was also missed by most of the ELISAs, had very faint staining of the gp160 and a very weak staining of p24, and would most probably be interpreted as negative. Adaption of currently available tests to anti-subtype O is needed for the future reliability of anti-HIV diagnostic reagents.

AIDS Serodiagnosis↗
PubMed1995

Nonoxynol-9 use, genital ulcers, and HIV infection in a cohort of sex workers.

OBJECTIVES: To measure the associations between use of nonoxynol-9 (N-9) and incidence of genital ulcers, and incident ulcers and HIV seroconversion. METHODS: In a study of barrier contraceptive use and HIV infection, 273 female sex workers used condoms and 100 mg N-9 suppositories, and recorded sexual activity on coital logs. Genital ulcers were diagnosed clinically at monthly clinic visits. HIV infection was diagnosed by ELISA and Western blot. We calculated ulcer incidence rates by level of N-9 use. A nested matched case-control analysis assessed the effect of ulcers on HIV acquisition. RESULTS: More frequent N-9 use was not associated with genital ulcers and may have been protective against the lesions. Ulceration was not a strong risk factor for HIV acquisition in this study (odds ratio 1.1; 95% confidence interval 0.3-3.5). CONCLUSIONS: Frequent use of N-9 can cause genital irritation and ulceration. Ulcers, in turn, may be risk factors for HIV acquisition. This study, however, did not find an association between N-9 use and ulcers, nor between ulcers and HIV. There is probably a threshold of N-9 use frequency or dose below which the risk of ulceration is minimal. Ulcers due to infectious causes may have been prevented by N-9 use in this cohort.

Adult↗
PubMed1994

Sensitivity of United States HIV antibody tests for detection of HIV-1 group O infections.

Infections by highly divergent strains of HIV-1, first detected in central Africa and grouped provisionally as group O, have not been reliably detected by certain European HIV screening tests. Serum specimens from eight probable group O infections from Cameroon were tested by ten HIV assays licensed by the US Food and Drug Administration. All assays based on synthetic peptides or recombinant antigens failed to detect at least one of the infections; assays based on whole-virus lysates performed better. Divergent HIV strains may be undetected by current HIV tests. Thus active surveillance for and characterisation of HIV variants to evaluate and, when necessary, modify current tests is urgently needed.

AIDS Serodiagnosis↗