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HIV-1 Viral Load and CD4 Assessment in HIV-1 Infected Pregnant Women Supported as Part of PMTCT in N’Djamena, Chad  [PDF]
Adoum Fouda Abderrazzack, Mounerou Salou, Yaovi Ameyapoh, Mahamat Nour Aguid, Bertin Tchombou Hig-Zounet, Adawaye Chatte, Abdelsalam Tidjani
World Journal of AIDS (WJA) , 2015, DOI: 10.4236/wja.2015.53027
Abstract: In Sub-Saharan Africa, HIV affects lots of women of childbearing age; without prevention they can transmit the virus to their child. A cross-sectional study was conducted in the center of Psycho Medico-Social Support (APMS) in N’Djamena, Chad from January 2014 to March 2015. Our sampling concerned HIV-1 infected pregnant women followed up for PMTCT and their newborn. CD4+ lymphocytes and HIV-1 viral load were tested respectively with PIMATM and Abbott m2000 Real Time in mothers. Early infant diagnosis of HIV-1 was done in Children using PCR tool (Abbott m2000 Real Time). Pregnant women included in the study had a median age of 25 years (IQR, 22 - 30 years). Most of them (75.6%) (34/45), were under combination ART (TDF + 3TC or FTC + EFV). The median duration on ART was 4 month (IQR [3 - 5 months]). Nevirapine syrup was administrated to newborns as prophylaxis at least for the first six weeks of life until EID was done. At ART initiation, mothers’ LTCD4+ median was 249 cells/mm3 (IQR: 95 - 674 cells/mm3). After a median duration of 4 months on ART, LTCD4+ median was 530 cells/mm3 (IQR [263 - 1220 cells/mm3]). Viral load assessment in mothers showed that 15.5% (7/45) were undetectable, 75.6% (34/45) were detectable with a VL < 3log copies/ml and 8.8% (4/45) at virologic failure (VL > 3log copies/ ml). Four (11.4%) of 35 children included were tested positive at EID for HIV-1. Antiretroviral treatment management in pregnant women can improve their health and reduce the risk of MTCT. Availability of virologic monitoring in routine is essential for pregnant women in resources limited setting for preventing HIV transmission to their new-born and keep them alive.
Implementation of an In-House Quantitative Real-Time PCR for Determination of HIV Viral Load in Kinshasa  [PDF]
Erick Ntambwe Kamangu, Adawaye Chatte, Raphael Boreux, Richard Lunganza Kalala, Georges Lelo Mvumbi, Patrick Demol, Dolores Vaira, Pierre Marie Hayette
Open Access Library Journal (OALib Journal) , 2014, DOI: 10.4236/oalib.1100855
Abstract: Background: Measurement of Viral Load (VL) is the most reliable mean for evaluating virological monitoring of the Human Immunodeficiency Virus (HIV) infection. It allows determination of the amount of virus present in a given volume. Due to the constraints of costs, the VL is not often requested for patient’s follow-up in countries with limited resources. Hence the objective of this study is to implement an in-house Quantitative Real-Time PCR to assess the VL of HIV infected patients in Kinshasa. Methods: One hundred and fifty five patients positive for HIV type 1, naive of Antiretroviral Therapy (ART) and eligible for treatment were included in the study. Five milliliter of blood was collected in a tube with anticoagulant. One milliliter of plasma was sent to the laboratory for analysis. After RNA extraction, a Quantitative Real time PCR was performed on a portion of the region of the Long Terminal Repeat (LTR) of the virus. Results: Of 155 samples received for determination of VL by Quantitative Real-Time PCR, 153 were successfully amplified according to the protocol. The median VL was 301052.97 copies/ml or 5.48 log10. Conclusions: The results of VL were used to assess the feasibility of the Real-Time Quantitative PCR. It turns a simple, reliable and less expensive alternative for the diagnosis and virological monitoring of HIV patients under ART.
Comparison of an In-House Quantitative Real-Time PCR and COBAS AmpliPrep/TaqMan Roche for Determination of Viral Load for HIV Type 1 Non-B  [PDF]
Erick Ntambwe Kamangu, Adawaye Chatte, Raphael Boreux, Fabrice Susin, Richard Lunganza Kalala, Georges Lelo Mvumbi, Patrick De Mol, Marie-Pierre Hayette, Dolores Vaira
Open Access Library Journal (OALib Journal) , 2015, DOI: 10.4236/oalib.1101402
Abstract: Context: The in-house techniques or experimental methods are increasingly recommended for their low-cost reagents for the determination of the Viral Load (VL) in resource-limited settings. The objective of this study was to compare the determination of VL from HIV-1 non-B samples by an in-house technique with the COBAS AmpliPrep/TaqMan version 2.0. Method: In this cross-sectional study, 39 plasma samples from patients infected with HIV type 1 non-B from N’Djamena and Kinshasa were used to determine the VL using the two techniques. Results: The mean values of VL are respectively 4.68 ± 1.26 and 4.58 ± 1.33 log10 RNA copies/ml for the COBAS AmpliPrep/TaqMan assays and the in-house assays. A good correlation (Spearman Correlation) was obtained, with a coefficient (R2) of 0.9452. Conclusion: This study demonstrates that there is no significant difference between the results of VL determined by the COBAS AmpliPrep/TaqMan assays and the in-house assays used.
Use of Dried Blood Spot to Improve the Diagnosis and Management of HIV in Resource-Limited Settings  [PDF]
Chatté Adawaye, Erick Kamangu, Ali Mahamat Moussa, Bertin Tchoumbou, Dolores Vaira, Michel Moutschen
World Journal of AIDS (WJA) , 2013, DOI: 10.4236/wja.2013.33033

Over 75% of people infected with HIV live in countries where health resources are very limited for the diagnosis and biological monitoring of people infected by the virus. In resource-limited settings, the use of DBS is a valuable alternative. It has provided technical and economical alternative to the collection of blood in the tubes for testing HIV infection. The DBS can be kept for over a year, it is economical in storage space and facilitates storage conditions because it can be stored at room temperature. It is more discreet and easier to carry over liquid samples that require tubes and other appropriate materials. The amount is sufficient for certain analyses of DNA generally, but may be insufficient for the analysis of viral RNA if the viral load is low. Its disadvantage is often associated with small amounts of blood collected available for testing, and the difficulties encountered in laboratories to extract the maximum possibilities without material contamination. DBS can be stored at room temperature (25 - 35), at 4, -20 or even -70. With PCR, the DBS is a suitable medium for the diagnosis of patients infected with HIV, virological monitoring by the VL and even analyzing viral genotype. It is a handy stand for the collection, transport and analyses of biological monitoring of HIV infection. It is indeed very suitable for environments with limited accessibility where it is difficult for specialized laboratories to monitor these patients. The DBS is suitable for resource-limited settings.

Immunovirologic Evaluation of Triomune (Lamivudine, Stavudine and Nevirapine) Antiretroviral Therapy in First Line HIV-1 Adult Patients in N’Djamena, Chad  [PDF]
Chatté Adawaye, Kamangu Erick, Soudy I. Djibrine, Aoudalkarim Moussa Chahad, Ali Mahamat Moussa, Tchombou HZ Bertin, Vaira Dolores, Moutschen Michel
World Journal of AIDS (WJA) , 2014, DOI: 10.4236/wja.2014.43035

Contexte: The fight against HIV/AIDS epidemics is one of the greatest challenges of this century. The epidemic affects generally under-developed countries, and Sub-Saharan Africa are the most concerned. The combined marketed form known as Triomune was used as first-line treatment in several sub-Saharan African Countries (60% of VIH infected people), including Chad. However, no evaluation has been done for that treatment in the country. Objective: To evaluate the efficacy and safety immuno-virological of Triomune at the General Hospital in N’Djamena/Chad. Methods: 48 HIV-1 positive patients eligible for ARV treatment were enrolled in our study, and they have been then followed for 8 months. We have measured in these patients the CD4 cell count before treatment and at the 8th month of treatment. After 8 months of treatment, we have also evaluated the Lymphocyte T CD4 and the plasma viral load (VL). Comparisons of means of CD4 lymphocytes and plasma CV (≥1000 copies/ml) were used to define treatment failure. Results: 48 patients were under Triomune regime. The average CD4 count was decreased from 462 ± 179.22 [56 - 981] cells/mm3 before treatment to 327.23 ± 153.77 [10 - 1008] cells/mm3 at the 8th month of treatment. The mean plasma viral load for patients was 66008.62 copies/ml. The failure rate to Triomune was 43.75% (21/48). Conclusion: Aside from the side effects already described for Triomune, our study reveals a high treatment failure rate. Hence, there is the need of regular revisions of therapeutic regime administer in the first intention.

Correlation between Sequencing Results from Liquid Plasma and Dried Plasma Spot (DPS) for Determination of HIV Type 1 Non-B Subtypes  [PDF]
Erick Ntambwe Kamangu, Adawaye Chatté, Dolores Vaira, Patrick de Mol, Georges Lelo Mvumbi, Richard Lunganza Kalala, Marie-Pierre Hayette
Open Access Library Journal (OALib Journal) , 2017, DOI: 10.4236/oalib.1102922
Background: The blotting paper is an alternative to the collection of blood in the tubes for analysis, especially in the field of Human Immunodeficiency Virus infection. This technique allows to easily send the collected samples to specialized laboratories while limiting the stresses of storage and transport. Objective: The objective of this study was to compare the results of sequencing performed on liquid plasma and Dried Plasma Spot (DPS) for the variants of HIV-1 non-B. Methodology: Fifty subjects diagnosed positive for HIV Type 1 using the Rapid Screening Tests voluntarily participated in this study. Two hundred microliters of plasma are deposited on blotting paper Whatman 903 and 500 μl in a micro tube. RNA was extracted from 140 μl of plasma fluid and from a piece of DPS of 5 mm of diameter using the QIAamp RNA Mini Kit QIAGEN. After extraction, the Viral Load (VL) was performed on each sample of liquid plasma. A Reverse Transcription PCR and Nested PCR were used to amplify the regions of interest on the Protease and Reverse Transcriptase for subsequent sequencing. Results: Protease and Reverse Transcriptase were amplified and sequenced respectively for 44 (88%) and 48 (96%) with the liquid plasma samples and 40 (80%) and 45 (90%) with the DPS. The results of Viral Loads were in the range of 2.5 log10 and 6.5 log10. The results of sequencing are comparable for plasma samples and DPS. The correlation coefficient (R2) between the two methods is good (R2 = 0.903, p < 0.001). Conclusion: Liquid Plasma and Dried Plasma Spot give highly correlated results for sequencing strains of HIV type 1 non-B.
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