The Diamniadio Institute for Health Research, Epidemiological Surveillance, and Training (Iressef): One Man's Vision and a Bet on the Future.

To reduce congestion in its capital, Dakar, Senegal decided in 2012 to construct a new city in Diamniadio, in the suburbs. This new urban center, an integral part of the Emerging Senegal Plan (PSE), is a major first step towards the organization of land use planning. The Institute for Health Research, Epidemiologic Surveillance and Training (Iressef) is one of the very first new operational buildings in this new city. Conceived and directed by Professor Souleymane Mboup, Iressef was made possible by the support of the government of Senegal and the GILEAD Foundation. The vision of its sponsors is to make it a hub of excellence for research in the domain of tropical infectious diseases, with expertise and technical equipment and facilities comparable to those in research institutes in developed countries. The existing platforms include, among others, several state-of-the-art laboratories, a health and demographic surveillance system covering a population of 30 000 inhabitants, a clinical trial center, a dynamic community participation program, and a training center. To achieve this vision in the short- and long-terms, Iressef has developed a strategic 5-year plan focusing on two principal objectives, that is: (i) to conduct research programs according to the strictest ethical standards, and (ii) to train an elite group of Senegalese and African scientists, competitive and capable of developing health research in Africa. To attain these objectives, international partners will play an essential role.

[Use of dried blood spots in early diagnosis of HIV-1 infection in children born to HIV-infected mothers as part of the prevention of mother-to-child transmission in Benin]. / Utilisation des prélèvements capillaires sur buvard dans le diagnostic précoce de l'infection à VIH-1 chez des enfants nés de mères infectées dans le cadre de la prévention de la transmission mère-enfant au Bénin.

The goal of this study was to evaluate using the molecular diagnosis, infection transmission rate of HIV in children born to HIV-1 positive mothers as part of the prevention of mother-to-child transmission (PMTCT) in Benin. The sample consisted of 524 dried blood spots (DBS) of children born to HIV-1 positive mothers, from 30 sites (PMTCT) taken between October 2009 and June 2010. The diagnosis of HIV-1 was performed by the qualitative detection of viral nucleic acids (RNA and DNA) in DBS on filter paper using the Abbott RealTime(®) HIV-1 Qualitative assay. We found that 51 DBS were positive (9.7%) and 473 were negative (90.3%). The failure rate of PMTCT among 420 mothers who received antiretroviral prophylaxis was 6.7% (28/420). This failure rate was significantly higher among children born to infected mothers on antiretroviral monotherapy than on triple therapy (HAART). The results of our study enrich the data in the literature on highly active antiretroviral chemoprophylaxis to reduce the transmission of HIV-1 from mother to child.

[The challenges of training in medical laboratories in Africa]. / Le défi de la formation dans le domaine du laboratoire de biologie clinique en Afrique.

Sub-Saharan Africa has a considerable deficit in laboratory facilities. For a decade, international and national public and private initiatives have multiplied to expand both the supply and quality of medical laboratories in Africa. By 2020, the World Health Organization, with as its main operator the African Society for Laboratory Medicine, will have provided training for 30,000 laboratory personnel and encouraged 2,500 laboratories to begin the accreditation process. In addition, the World Health Organization recommendations for treatment and care of HIV-infected individuals in resource-limited settings, revised in 2013, emphasize the need for laboratory monitoring to guide antiretroviral therapy. The University Diploma in Biological Retrovirology at the Cheikh Anta Diop University in Dakar, Senegal, offers multidisciplinary training in French at the postgraduate level in the complex and diverse field of biological monitoring of HIV infection in Africa. In nearly 10 years, more than 200 African biologists have been trained.

Tenofovir-emtricitabine-efavirenz in HIV-I-infected adults in Senegal: a 96-week pilot trial in treatment-naive patients.

We report the results of a pilot open-label trial of a tenofovir (TDF)/emtricitabine (FTC)/efavirenz (EFV) combination conducted in Dakar, Senegal. Forty HIV-1-infected patients, naive of antiretroviral treatment and without active opportunistic disease, were included and followed through 96 weeks. At weeks 48 and 96, respectively, 82.5% and 85% of patients had HIV-1 RNA <400 copies/mL (72.5% and 77.5% with HIV-1 RNA <50 copies/mL). Between baseline and week 96, the mean (SD) CD4 count increased from 126 (102) to 338 (155) cells/mm(3). The mean (SD) creatinine clearance decreased from 92 (36) to 73 (19) mL/min (P = .001). Treatment adherence was at least 94% at all scheduled visits. The efficacy and tolerability of a TDF/FTC/EFV combination were high and similar to those observed in Northern countries. This drug combination can be recommended in limited-resource countries, as did the World Health Organization (WHO) and should be made readily available as a fixed-dose combination.

[Decentralization of the immunological monitoring of people living with HIV/AIDS in a limiting setting with a low HIV seroprevalence: the experience of Senegal]. / Décentralisation du suivi immunologique des personnes vivant avec le VIH dans un pays à ressources limitées et à prévalence faible: expérience du Sénégal.

Our work aimed to propose a manual method of counting CD4 T lymphocytes which is an alternative magnetic immunoseparation followed by a reading with a fluorescence microscope as an alternative to the automated flow cytometry. This alternative technique is easier for use, less expensive and could answer the difficulties encountered for the monitoring CD4 T cells count in developing countries. The specific objectives were: 1) to train the technicians of the peripheral sites in order to make the numeration of the CD4 T lymphocytes more accessible at the peripheral level; 2) to equip the sites with necessary facilities for the T lymphocytes CD4 count; 3) to put in place a system of quality control permitting the reliability of the results. A hundred and fifty patients have been enrolled in three care services for people living with HIV/AIDS in Dakar. This population was constituted of 119 seropositive and 31 seronegative patients acting as control group to have some patients with high rates of T lymphocytes CD4. For the follow-up at peripheral level, the patients were constituted of the active line of the patients living with HIV/AIDS supported in the targeted sites. The measurements allowed studying concordances for different rates of lymphocytes: 0 to 199, 200 to 499 and over 500 cells by mm3. The results showed also a very good correlation (r = 0.97 or r = 0.98 according to the operator) between the two methods for CD4 rates inferior to 500 cells by mm3 among both the negative group and the HIV positive patients. We also discussed the profit of decentralization for the program and the patient, as well as the setting up of an external quality control to validate the alternative technique. According to the results, the Dynabeads is well correlated with the Facscount. It is a technique that can be used as an alternative in the zones with limited resources, low prevalence and for a small number of samples.

Hepatitis B, C seroprevalence and delta viruses in HIV-1 Senegalese patients at HAART initiation (retrospective study).

The aim of this study was to determine hepatitis co-infection in a cohort of HIV infected patients at their inclusion in the Senegalese Initiative of ART Access. B, C, and D Hepatitis viruses serological markers were checked retrospectively on 363 stored plasma. For HBV, the Abbott laboratories equipment IMx was used to detect HBs Ag and anti Core Ab on negative HBs Ag samples. For HDV, anti Delta Ab was performed using the Abbott Murex Kit on all HBs Ag positive samples. For HCV, anti HCV Ab was detected by IMx as double screening test and confirmed by INNO-LIA(TM) HCV Core of Innogenetics laboratories. The statistical analysis was done with STATA V8. The study population was composed of 164 men and 199 women aged between 16 and 66 years. The immune and virological markers averages at their enrollment were 154 cell/mm(3) for TLCD4+ (n = 355 patients) and 4.9 log for viral load (n = 277 patients). HBs Ag was found in 61 patients or 16.8% and the prevalence of anti-HBc Ab was 83.2% (252/295). 2 patients or 3% on HBs Ag positive sample presents HBV/HDV co-infection Ab anti HCV was detects in 6 patients or 1.6% after confirmation and 2 patients had triple infection with HBV. These results showed that the prevalence of HBV and HCV in the population of persons living with HIV/AIDS in Senegal is similar to that found in the general population. Our data indicated that hepatitis pathology in the PLwHIV was essentially due to HBV. Further studies are needed to diagnose occult hepatitis in order to set up therapeutic strategies taking into account co-infections by hepatitis viruses in the ART programmes.

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.

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.

Most env and gag subtype A HIV-1 viruses circulating in West and West Central Africa are similar to the prototype AG recombinant virus IBNG.

The genetic subtype was identified in gag and env of 219 HIV-1-positive samples collected in different African countries, 44 from Senegal, 55 from Cameroon, 82 from Gabon, and 38 from Djibouti. In total, 20 (9.1%) samples had discordant subtypes between gag and env, 6 of 44 (13.9%) in Senegal, 4 of 55 (7.2%) in Cameroon, 1 of 38 (2.6%) in Djibouti, and 10 of 82 (12.1%) in Gabon. Subtypes A and G were predominantly involved in the recombination events. Phylogenetic tree analysis of gag showed that an important number of the A sequences form a distinct subcluster with the AG-IBNG prototype strain (a complex A/G mosaic virus): 27 of 32 (84.3%) in Senegal, 12 of 17 (70.6%) in Nigeria, 24 of 39 (61.5%) in Cameroon, and 38 of 70 (54.3%) in Gabon. Full-length genome analysis of 3 and additional sequences in pol for 10 such strains confirmed that they have a similar complex A/G mosaic genomic structure. These data suggest that in West Africa, most probably between 60% and 84% of the subtype A viruses are recombinant AG-IBNG viruses. This finding has potential implications on future vaccine, diagnostic, and treatment strategies. The actual and future role of these viruses in the global pandemic must be monitored in all new molecular epidemiologic studies, a discrimination between subtype A and AG-IBNG-like viruses is necessary.

Identification of all HIV type 1 group M subtypes in Senegal, a country with low and stable seroprevalence.

A total of 343 HIV-1-positive samples obtained between June 1996 and March 1999 was genetically characterized in the envelope region by HMA and/or sequencing. The env subtype distribution was as follows: 290 (84.6%) A, 22 (6.5%) B, 16 (4.7%) C, 8 (2.5%) D, 1 (0.03%) E, 1 (0.03%) F1, 4 (1.2%) G, and 1 (0.03%) H. For 77 samples the p24 region from the gag gene was also sequenced, and for 9 (11.6%) the subtypes between env and gag were different. Phylogenetic tree analysis showed the predominance of AG-IBNG-like viruses among gag and env subtype A sequences. HMA is relatively simple and requires less sophisticated technical facilities compared with sequencing, and in Senegal 323 (94.2%) of the 343 samples could be identified by this technique. However, in the actual configuration of the assay, discrimination between the recombinant AG-IBNG-like recombinant viruses, which are predominant in Senegal, and the nonrecombinant subtype A viruses is not possible.