Diagnostic moléculaire du Cytomégalovirus (CMV), de l’herpès virus humain de type 6 (HHV6) et d’Epstein-Barr virus (EBV) par PCR en temps réel chez les femmes enceintes VIH séropositives et séronégatives à Ouagadougou, Burkina Faso

Introduction: les herpès virus EBV, CMV et HHV-6 sont des virus qui évoluent sous le modèle pandémique et sont responsables d’infections congénitales pouvant provoquer des séquelles graves chez les nouveau-nés. L’objectif de cette étude était de déterminer les prévalences de CMV, EBV et HHV-6 chez les femmes enceintes VIH(+) et VIH(-) à Ouagadougou. Méthodes: dans cette étude 200 échantillons de plasma sanguin de femmes enceintes dont 100 femmes VIH(+) et 100 femmes VIH(-) ont été diagnostiqués par PCR multiplex en temps réel pour les trois infections (EBV, CMV et HHV-6). Résultats: sur l’ensemble des 200 échantillons analysés, 18 (9,0%) étaient positifs à au moins un des trois virus, 12 (6,0%) étaient positifs au EBV, 13 (6,5%) au CMV et 12 (6,0%) positifs au HHV-6. Parmi les 18 cas d’infections, nous avons trouvé 10 cas (55,6%) de coïnfections dont 90,0% (9/10) d’infection multiple EBV/CMV/HHV6 et 10,0% de coinfection EBV/HHV6. Le taux d’infection HHVs était plus élevé chez les femmes VIH(-) que celles VIH(+) (12,0% versus 6,0%). Parmi les VIH(+), la PCR a révélé 7,1% (soit 6/85) d’infection HHVs chez celles qui n’étaient pas sous ARV contre 0% chez celles sous ARV. Conclusion: les herpès virus sont fréquents chez les femmes enceintes au Burkina Faso et pourraient constituer une menace chez ces dernières à cause des complications et des risques d’infection pour le nouveau-né.The Pan African Medical Journal 2016;2

Molecular diagnosis of Shigella, Salmonella and Campylobacter by multiplex Real-time PCR in stool culture samples in Ouagadougou (Burkina Faso)

ABSTRACT:Background: Bacteriological diagnosis of Campylobacter spp, Salmonella spp and Shigella spp could be necessary in the case of infectious gastroenteritis syndrome.The objective of this study was to diagnose concomitantly the three enteropathogenic bacteria by multiplex Real-Time PCR in stool culture samples in Ouagadougou (Burkina Faso).Materials and Methods: The study was conducted from February 5th to March 9th, 2013. Two hundred stool samples were received during the study period. The bacteria were identified by bacterial culture following by multiplex Real-Time PCR.Results: Shigella spp and Campylobacter spp were sought by culture in all 200 samples. Enteropathogenic E. coli was sought only in 37 samples from all children under 2 years old. The bacterial culture was positive in 12 stool samples. Shigella spp and Salmonella spp. were isolated respectively in 5 (2.5%) and 3 samples (1.5%). Enteropathogenic E. coli was isolated in 10.8% (4/37) of the samples tested.The multiplex real-time PCR identified bacteria in 20 patients, including 17 cases of Shigella spp., 1 case of Salmonella spp. and 2 cases of Campylobacter spp.Conclusions: This study has highlighted the low frequency of 3 sought bacterial genera in stool samples. It has also demonstrated a significant difference between the culture and the multiplex Real-Time PCR method in the diagnosis of Shigella

Human immunodeficiency virus type 1 drug resistance in a subset of mothers and their infants receiving antiretroviral treatment in Ouagadougou, Burkina Faso

The emergence of HIV-1 drug resistance (HIVDR) is a public health problem that affects women and children. Local data of HIVDR is critical to improving their care and treatment. So, we investigated HIVDR in mothers and infants receiving antiretroviral therapy (ART) at Saint Camille Hospital of Ouagadougou, Burkina Faso. This study included 50 mothers and 50 infants on ART. CD4 and HIV-1 viral load were determined using FACSCount and Abbott m2000rt respectively. HIVDR was determined in patients with virologic failure using ViroSeq HIV-1 Genotyping System kit on the 3130 Genetic Analyzer. The median age was 37.28 years in mothers and 1.58 year in infants. Sequencing of samples showed subtypes CRF02_AG (55.56%), CRF06_cpx (33.33%) and G (11.11%). M184V was the most frequent and was associated with highlevel resistance to 3TC, FTC, and ABC. Other mutations such as T215F/Y, D67N/E, K70R, and K219Q were associated with intermediate resistance to TDF, AZT, and 3TC. No mutation to LPV/r was detected among mothers and infants. The findings of HIVDR in some mothers and infants suggested the change of treatment for these persons

Diagnostic biologique différentiel entre le paludisme et la dengue chez des patients fébriles à Ouagadougou au Burkina Faso dans un contexte d’endémie des deux maladies

La dengue constitue un problème de santé publique au Burkina Faso. Notre objectif était de faire le diagnostic biologique de la dengue chez des patients fébriles. Cette étude a porté sur 204 patients. Les kits « Dengue Duo de SD Bioline » ont été utilisés pour le dépistage. Une recherche de Plasmodium par la méthode de goutte épaisse a été réalisée chez les patients ainsi que le dosage du taux de CRP. La RT- PCR en temps réel a été utilisée pour confirmer les résultats positifs au test rapide. Environ 25,98 % des patients étaient TDR positif. L'étude a confirmé la présence de DENV chez 3,77 % des patients avec une association statistiquement significative entre le taux de plaquettes et la positivité à l’Ag NS1. Environ 87,87 % des patients qui avaient des gouttes épaisses positives présentaient également des taux de CRP élevés comparativement aux patients qui avaient des taux de CRP normaux et des gouttes épaisses positives (7,01 %) avec une différence statistiquement significative. Dans cette étude, nous proposons des pistes de diagnostic différentiel entre la dengue et le paludisme, dans un contexte d’endémie des deux maladies, à partir de paramètres biologiques tels que les taux de plaquettes et de CRP.Mots-clés: Dengue, Diagnostic, RT-PCR, Burkina FasoEnglish Title: Differential biological diagnosis between malaria and dengue in febrile patients in Ouagadougou, Burkina Faso in a context of endemic diseaseEnglish AbstractDengue fever is a public health problem in Burkina Faso. The goal of this study was to make the biological diagnosis of dengue in clinically suspect patients. This study involved 204 clinically suspected dengue patients. Dengue Duo kits from SD Bioline were used for screening. Plasmodium was investigated by the thick-film method in all patients included as well as the dosage of CRP. Complementary biological analyzes were carried out. Real-time RT-PCR was used to confirm positive results in the rapid test. Rapid tests revealed a 25.98% positive Dengue rate. The study confirmed by RT-PCR the effective presence of DENV in 3.77% of patients. Our study revealed a statistically significant association (p = 0.046) between platelet count and NS1 Ag positivity. In contrast, 87.87% (29/33) of patients who had malaria also had high CRP levels compared to patients with normal CRP and thick positive seizures (7.02%) with a statistically significant difference (p = 0.018). In this study, we confirm the responsibility of dengue arboviruses in cases of disease in our country, but also we propose ways of differential diagnosis between dengue and malaria, in an endemic context of both diseases, based on biological parameters such as platelet and CRP levels.Keywords: DENV, Diagnostic, RT-PCR, Burkina Fas

Glucose-6-phosphate dehydrogenase (G6PD) deficiency is associated with asymptomatic malaria in a rural community in Burkina Faso

Objective: To investigate 4 combinations of mutations responsible for glucose-6-phosphate dehydrogenase (G6PD) deficiency in a rural community of Burkina Faso, a malaria endemic country. Methods: Two hundred individuals in a rural community were genotyped for the mutations A376G, G202A, A542T, G680T and T968C using TaqMan single nucleotide polymorphism assays and polymerase chain reaction followed by restriction fragment length polymorphism. Results: The prevalence of the G6PD deficiency was 9.5% in the study population. It was significantly higher in men compared to women (14.3% vs 6.0%, P=0.049). The 202A/376G G6PD A- was the only deficient variant detected. Plasmodium falciparum asymptomatic parasitaemia was significantly higher among the G6PD-non-deficient persons compared to the G6PD-deficient (P<0.001). The asymptomatic parasitaemia was also significantly higher among G6PD non-deficient compared to G6PD-heterozygous females (P<0.001). Conclusions: This study showed that the G6PD A- variant associated with protection against asymptomatic malaria in Burkina Faso is probably the most common deficient variant

MOLECULAR HETEROGENEITY OF GLUCOSE-6-PHOSPHATE DEHYDROGENASE DEFICIENCY IN BURKINA FASO: G-6-PD BETICA SELMA AND SANTAMARIA IN PEOPLE WITH SYMPTOMATIC MALARIA IN OUAGADOUGOU

The G-6-PD deficiency has an important polymorphism with genotypic variants such as 202A/376G, 376G/542T and 376G/968T known in West African populations. It would confer protection against severe forms of malaria although there are differences between the various associations in different studies. In this study we genotyped six (06) variants of the G-6-PD gene in people with symptomatic malaria in urban areas in Burkina Faso. One hundred and eighty-two (182) patients who tested positive using rapid detection test and microscopy were included in this study. A regular PCR with the GENESPARK G6PD African kit was run followed by electrophoresis, allowing initially to genotype six SNPs (G202A, A376G, A542T, G680T, C563T and T968C). Women carrying the mutations 202A and/or 376G were further typed by real-time PCR using TaqMan probes rs1050828 and rs1050829. In the study population the G-6-PD deficiency prevalence was 9.9%. In addition of G-6-PD A- (202A/376G) variants, 376G/542Tand 376G/968T were detected. Hemoglobin electrophoresis revealed that 22.5% (41/182) of the individuals had HbAC compared with 2.2% with HbAS and one individual had double heterozygous HbSC. There was no correlation between the G-6-PD deficiency or haemoglobinopathies and symptomatic malaria in this study. As opposed to previous genotyping studies carried out in Burkina Faso, this study shows for the first time the presence of the variant A- (376G/968C) and warrants further investigation at the national level and in specific ethnic groups