The reliability of sickling and solubility tests and peripheral blood film method for sickle cell disease screening at district health centers in Uganda.

Bien que les analyses hématies falciformes, la solubilité et le film sanguin périphérique soient aujourd’hui disponibles pour le dépistage de la maladie drépanocytaire en Ouganda, leur fiabilité et aisance d’applicabilité n’ont pas été déterminées. Cette étude a été, par conséquent réalisée pour déterminer la fiabilité de la méthode de dépistage des analyses des hématies falciformes et de la solubilité ainsi que le film sanguin périphérique pour la SCD (sigle anglais pour Sickle cell disease) en Ouganda. Ceci a été une étude descriptive basée au laboratoire et réalisée par le Collège des Sciences de la Sante de l’Université de Makerere. Les 200 prélèvements des enfants ages de 6 mois à 5 ans ont été analysés de façon indépendante en utilisant la méthode des analyses d’hématies falciformes, la solubilité et le film sanguin périphérique. L’acétate de cellulose de l’électrophorèse de l’hémoglobine a été utilise comme échantillon. Les analyses des hématies falciformes et de solubilité ont eu la sensibilité de 65,0% et 45,0%, respectivement, et le film sanguin périphérique a eu 35,0%/. Les hématies falciformes, la solubilité et le film sanguin périphérique ont eu les spécifications de 95,6%, 90,0% et 96,7% respectivement. Les hématies falciformes ont eu la précision diagnostique de 92,5%, la solubilité – 85,5% et le film sanguin périphérique – 90,5%. Les hématies falciformes ont eu le Kappa de Cohen de 0,6, la solubilité – 0,3 et le film sanguin périphérique – 0,4. L’analyse des hématies falciformes a eu une période de rotation de 38 minutes, la solubilité – 70 minutes et le film sanguin périphérique – 44 minutes. En conclusion, l’analyse des hématies falciformes a été plus fiable et plus facile à exécuter que l’analyse de la solubilité et la méthode de film sanguin périphérique. Elle serait, par conséquent, une analyse recommandée pour le dépistage préliminaire des enfants pour la SCD aux centres de santé IV au niveau de districts et confirmer seulement les résultats positifs en utilisant l’électrophorèse de l’hémoglobine.KEY WORDS: Screening - Sickling and solubility tests -Peripheral blood film - Sickle cell disease - Uganda

Determination of LDL-cholesterol: direct measurement by homogeneous assay versus Friedewald calculation among Makerere University undergraduate fasting students

The treatment of patients for coronary heart disease risk requires knowledge of the plasma lipid levels. Low density lipoprotein cholesterol (LDL) levels make a strong basis for therapeutic decisions. Although there are incongruities among values of LDL from different methods of determining LDL, the clinician is not routinely informed of the method used. The purpose of this study was to compare LDL levels determined by the Friedewald equation with those assayed by the Kyowa Madox method. The lipid results previously measured by Kyowa Madox method among Makerere University fasting students and reported earlier wereretrieved. The measured values of total cholesterol (TC), High Density Lipoprotein cholesterol (HDL) and triacylglycerols (TG) were used to calculate LDL using Friedewald equation in which LDL= TC-HDL-TG/2.2mmol/L. The values obtained were compared non parametrically with the assayed values previously reported. Our results showed a high value of correlation between measured and calculated LDL so that in general, the two methods can be used interchangeably in this population. However, in cases of dyslipidaemia, the calculated values tend to be lower than the assayed values. It is therefore recommended that clinical laboratories should report the LDL values along with the determination method used, the alert values, the reference ranges, the desirable ranges and the therapeutic targets. © 2010 International Formulae Group. All rights reserved.Keywords: Homogeneous assay, LDL cholesterol, direct measurement, Friedewald equation, comparison

Plasma levels of DDT/DDE and liver function in malaria control personnel 6 months after indoor residual spraying with DDT in northern Uganda, 2008

Objective. We investigated the relationship between plasma levels of dichlorodiphenyltrichloroethane (DDT) and liver function in malaria control personnel 6 months after one round of DDT indoor residual spraying (IRS). Method. This was a cross-sectional study in the districts of Apac and Oyam of Lango, northern Uganda. Volunteers were clinically examined, and 5 ml samples of venous blood were taken in heparinised tubes for a 6-month post-spray screening for DDT and plasma markers of liver function and internal organ disease. DDE/DDT was assayed using ELISA kits (Abraxis, USA); plasma enzyme activity concentrations of amylase, aspartate aminotransferase (AST), alanine aminotransferase (ALT) and gamma glutamyl transpeptidase (GGT) were analysed using routine clinical chemistry-automated methods (Konelab, Vantaa, Finland). Results. All 96 plasma samples analysed for xenobiotics contained DDE/DDT in the empirical range of 24.00 - 128.00 parts per billion (ppb) with a mean (SD) of 77.00 (±26.00) ppb. All 119 plasma samples studied for the markers exhibited enzyme activity concentration values within the population reference ranges, with empirical means (SD) of amylase 71.86 (34.07), AST 23.83 (12.71), ALT 7.84 (10.01) and GGT 58.37 (62.68) µg/l. Conclusion. Six months after IRS with DDT, the spray team had an average concentration of plasma DDE/DDT of 77 ppb. This had no deleterious effect on liver function. We recommend continued use of DDT for IRS disease control in Uganda until better practical alternatives are available

Quantitative Assessment of the Sensitivity of Various Commercial Reverse Transcriptases Based on Armored HIV RNA

The in-vitro reverse transcription of RNA to its complementary DNA, catalyzed by the enzyme reverse transcriptase, is the most fundamental step in the quantitative RNA detection in genomic studies. As such, this step should be as analytically sensitive, efficient and reproducible as possible, especially when dealing with degraded or low copy RNA samples. While there are many reverse transcriptases in the market, all claiming to be highly sensitive, there is need for a systematic independent comparison of their applicability in quantification of rare RNA transcripts or low copy RNA, such as those obtained from archival tissues.We performed RT-qPCR to assess the sensitivity and reproducibility of 11 commercially available reverse transcriptases in cDNA synthesis from low copy number RNA levels. As target RNA, we used a serially known number of Armored HIV RNA molecules, and observed that 9 enzymes we tested were consistently sensitive to ∼1,000 copies, seven of which were sensitive to ∼100 copies, while only 5 were sensitive to ∼10 RNA template copies across all replicates tested. Despite their demonstrated sensitivity, these five best performing enzymes (Accuscript, HIV-RT, M-MLV, Superscript III and Thermoscript) showed considerable variation in their reproducibility as well as their overall amplification efficiency. Accuscript and Superscript III were the most sensitive and consistent within runs, with Accuscript and Superscript II ranking as the most reproducible enzymes between assays.We therefore recommend the use of Accuscript or Superscript III when dealing with low copy number RNA levels, and suggest purification of the RT reactions prior to downstream applications (eg qPCR) to augment detection. Although the results presented in this study were based on a viral RNA surrogate, and applied to nucleic acid lysates derived from archival formalin-fixed paraffin embedded tissue, their relative performance on RNA obtained from other tissue types may vary, and needs future evaluation

Incidence and predictors of hospital readmission in children presenting with severe anaemia in Uganda and Malawi: a secondary analysis of TRACT trial data

Background: Severe anaemia (haemoglobin < 6 g/dL) is a leading cause of recurrent hospitalisation in African children. We investigated predictors of readmission in children hospitalised with severe anaemia in the TRACT trial (ISRCTN84086586) in order to identify potential future interventions. Methods: Secondary analyses of the trial examined 3894 children from Uganda and Malawi surviving a hospital episode of severe anaemia. Predictors of all-cause readmission within 180 days of discharge were identified using multivariable regression with death as a competing risk. Groups of children with similar characteristics were identified using hierarchical clustering. Results: Of the 3894 survivors 682 (18%) were readmitted; 403 (10%) had ≥2 re-admissions over 180 days. Three main causes of readmission were identified: severe anaemia (n = 456), malaria (n = 252) and haemoglobinuria/dark urine syndrome (n = 165). Overall, factors increasing risk of readmission included HIV-infection (hazard ratio 2.48 (95% CI 1.63–3.78), p < 0.001); ≥2 hospital admissions in the preceding 12 months (1.44(1.19–1.74), p < 0.001); history of transfusion (1.48(1.13–1.93), p = 0.005); and missing ≥1 trial medication dose (proxy for care quality) (1.43 (1.21–1.69), p < 0.001). Children with uncomplicated severe anaemia (Hb 4-6 g/dL and no severity features), who never received a transfusion (per trial protocol) during the initial admission had a substantially lower risk of readmission (0.67(0.47–0.96), p = 0.04). Malaria (among children with no prior history of transfusion) (0.60(0.47–0.76), p < 0.001); younger-age (1.07 (1.03–1.10) per 1 year younger, p < 0.001) and known sickle cell disease (0.62(0.46–0.82), p = 0.001) also decreased risk of readmission. For anaemia re-admissions, gross splenomegaly and enlarged spleen increased risk by 1.73(1.23–2.44) and 1.46(1.18–1.82) respectively compared to no splenomegaly. Clustering identified four groups of children with readmission rates from 14 to 20%. The cluster with the highest readmission rate was characterised by very low haemoglobin (mean 3.6 g/dL). Sickle Cell Disease (SCD) predominated in two clusters associated with chronic repeated admissions or severe, acute presentations in largely undiagnosed SCD. The final cluster had high rates of malaria (78%), severity signs and very low platelet count, consistent with acute severe malaria. Conclusions: Younger age, HIV infection and history of previous hospital admissions predicted increased risk of readmission. However, no obvious clinical factors for intervention were identified. As missing medication doses was highly predictive, attention to care related factors may be important. Trial registration: ISRCTN ISRCTN84086586. Keywords: Severe anaemia, Readmissio

Health Professional Training and Capacity Strengthening Through International Academic Partnerships: The First Five Years of the Human Resources for Health Program in Rwanda

Abstract Background: The Rwanda Human Resources for Health Program (HRH Program) is a 7-year (2012-2019) health professional training initiative led by the Government of Rwanda with the goals of training a large, diverse, and competent health workforce and strengthening the capacity of academic institutions in Rwanda. Methods: The data for this organizational case study was collected through official reports from the Rwanda Ministry of Health (MoH) and 22 participating US academic institutions, databases from the MoH and the College of Medicine and Health Sciences (CMHS) in Rwanda, and surveys completed by the co-authors. Results: In the first 5 years of the HRH Program, a consortium of US academic institutions has deployed an average of 99 visiting faculty per year to support 22 training programs, which are on track to graduate almost 4600 students by 2019. The HRH Program has also built capacity within the CMHS by promoting the recruitment of Rwandan faculty and the establishment of additional partnerships and collaborations with the US academic institutions. Conclusion: The milestones achieved by the HRH Program have been substantial although some challenges persist. These challenges include adequately supporting the visiting faculty; pairing them with Rwandan faculty (twinning); ensuring strong communication and coordination among stakeholders; addressing mismatches in priorities between donors and implementers; the execution of a sustainability strategy; and the decision by one of the donors not to renew funding beyond March 2017. Over the next 2 academic years, it is critical for the sustainability of the 22 training programs supported by the HRH Program that the health-related Schools at the CMHS significantly scale up recruitment of new Rwandan faculty. The HRH Program can serve as a model for other training initiatives implemented in countries affected by a severe shortage of health professionals

Knowledge gaps, attitude and beliefs of the communities about sickle cell disease in eastern and western Uganda

Background: The management of sickle cell disease (SCD) has remained insurmountable in developing countries such as Uganda, because most communities are not aware of it.Objective: To determine knowledge gaps, attitudes and beliefs of the communities about sickle cell disease in Eastern and Western Uganda.Design: Cross sectional descriptive study.Setting: The districts of Sironko and Mbale in Eastern Uganda and Mbarara and Ntungamo in Western Uganda.Subjects: Households, students and health workers.Results: Household respondents from Eastern Uganda were more aware of SCD than those from Western (