MOESM1 of Effectiveness of malaria control interventions in Madagascar: a nationwide case–control survey

Additional file 1. Effectiveness of LLIN in all areas with LLIN distribution. Bi- and multivariate analyses of risk factors for developing a clinical malaria episode,including bed net use

Dried-Blood Spots: A Cost-Effective Field Method for the Detection of Chikungunya Virus Circulation in Remote Areas

<div><p>Background</p><p>In 2005, there were outbreaks of febrile polyarthritis due to Chikungunya virus (CHIKV) in the Comoros Islands. CHIKV then spread to other islands in the Indian Ocean: La Réunion, Mauritius, Seychelles and Madagascar. These outbreaks revealed the lack of surveillance and preparedness of Madagascar and other countries. Thus, it was decided in 2007 to establish a syndrome-based surveillance network to monitor dengue-like illness.</p><p>Objective</p><p>This study aims to evaluate the use of capillary blood samples blotted on filter papers for molecular diagnosis of CHIKV infection. Venous blood samples can be difficult to obtain and the shipment of serum in appropriate temperature conditions is too costly for most developing countries.</p><p>Methodology and principal findings</p><p>Venous blood and dried-blood blotted on filter paper (DBFP) were collected during the last CHIKV outbreak in Madagascar (2010) and as part of our routine surveillance of dengue-like illness. All samples were tested by real-time RT-PCR and results with serum and DBFP samples were compared for each patient. The sensitivity and specificity of tests performed with DBFP, relative to those with venous samples (defined as 100%) were 93.1% (95% CI:[84.7–97.7]) and 94.4% (95% CI:[88.3–97.7]), respectively. The Kappa coefficient 0.87 (95% CI:[0.80–0.94]) was excellent.</p><p>Conclusion</p><p>This study shows that DBFP specimens can be used as a cost-effective alternative sampling method for the surveillance and monitoring of CHIKV circulation and emergence in developing countries, and probably also for other arboviruses. The loss of sensitivity is insignificant and involved a very small number of patients, all with low viral loads. Whether viruses can be isolated from dried blood spots remains to be determined.</p></div

Sensitivity and specificity of test performed on DBFP for the diagnosis of Chikungunya virus infection^a.

a<p>Sensitivity, 93.1% (68/73, (95% CI:[84.7–97.7]); specificity, 94.4% (102/108, (95% CI:[88.3–97.7]); Kappa coefficient, 0.87 (95% CI:[0.80–0.94]).</p

Ct Differences in RNA amplification from 15 µl aliquots of whole blood and DBFP.

<p>Ct Differences in RNA amplification from 15 µl aliquots of whole blood and DBFP.</p

Primers and probes used in amplification of Chikungunya RNA as previously described by Laurent et al. 2007 [28] (modifications of primers are indicated in bold).

<p>Primers and probes used in amplification of Chikungunya RNA as previously described by Laurent et al. 2007 <a href="http://www.plosntds.org/article/info:doi/10.1371/journal.pntd.0002339#pntd.0002339-Laurent1" target="_blank">[28]</a> (modifications of primers are indicated in bold).</p

Incidence of ILI during each regional peak.

<p>Incidence of ILI during each regional peak.</p

Flow charts of cases from visits to influenza results.

<p>Flow charts of cases from visits to influenza results.</p

A(H1N1)pdm09 diffusion in Madagascar-the “peaks road.”

<p>A(H1N1)pdm09 diffusion in Madagascar-the “peaks road.”</p

Distribution of overall visits, fever-related illnesses, influenza-1-like illnesses, and positive results by age.

*<p>comparison of A(H1N1)pdm positivity by age using logistic regression analysis.</p

Study on causes of fever in primary healthcare center uncovers pathogens of public health concern in Madagascar

<div><p>Background</p><p>The increasing use of malaria diagnostic tests reveals a growing proportion of patients with fever but no malaria. Clinicians and health care workers in low-income countries have few tests to diagnose causes of fever other than malaria although several diseases share common symptoms. We propose here to assess etiologies of fever in Madagascar to ultimately improve management of febrile cases.</p><p>Methodology</p><p>Consenting febrile outpatients aged 6 months and older were recruited in 21 selected sentinel sites throughout Madagascar from April 2014 to September 2015. Standard clinical examinations were performed, and blood and upper respiratory specimens were taken for rapid diagnostic tests and molecular assays for 36 pathogens of interest for Madagascar in terms of public health, regardless of clinical status.</p><p>Principal findings</p><p>A total of 682 febrile patients were enrolled. We detected at least one pathogen in 40.5% (276/682) of patients and 6.2% (42/682) with co-infections. Among all tested patients, 26.5% (181/682) had at least one viral infection, 17.0% (116/682) had malaria and 1.0% (7/682) presented a bacterial or a mycobacterial infection. None or very few of the highly prevalent infectious agents in Eastern Africa and Asia were detected in this study, such as zoonotic bacteria or arboviral infections.</p><p>Conclusions</p><p>These results raise questions about etiologies of fever in Malagasy communities. Nevertheless, we noted that viral infections and malaria still represent a significant proportion of causes of febrile illnesses. Interestingly our study allowed the detection of pathogens of public health interest such as Rift Valley Fever Virus but also the first case of laboratory-confirmed leptospirosis infection in Madagascar.</p></div