Status of Onchocerciasis Elimination in Gabon and Challenges: A Systematic Review.

Control and treatment programs (CDTI) have been set up nationally in all endemic countries to overcome the impact of onchocerciasis on the affected populations. However, Gabon must still succeed in setting up real onchocerciasis control programs. Here, various database articles have been used to provide the scientific community with a summary document showing the mapping of this disease in Gabon. The articles dealing with onchocerciasis, animal reservoirs, surveillance, and elimination were analyzed. Results showed that little research has been performed. Most studies are concentrated in one region (The area of Lastourville). In addition, we observed that the distribution of the disease varies significantly across the country. Indeed, specific environments present a hyper-endemicity of the disease, while others are meso and hypo-endemic. So, we found some departments with a prevalence ranging from 0% to over 20%; within them, villages had infection levels comprising 10% to 60%, indicating potential hotspots. Vectors activities were studied in some areas. This paper showed the challenges encountered in the country to eliminate this disease. One solution is a deeper understanding of the disease's bioecology to establish effective health policies to eliminate onchocerciasis in Gabon effectively.

Comparison of six methods for Loa loa genomic DNA extraction.

OBJECTIVES: Good-quality and sufficient DNA is essential for diagnostics and vaccine development. We aimed to compare six DNA extraction techniques applied to Loa loa microfilariae in order to evaluate the purity and integrity of extracts in terms of quality and quantity. METHODS: The microfilariae were purified via a Percoll gradient procedure with blood from hyper-microfilaremic individuals (> 30,000 microfilaria [mf]/ml). DNA extraction was carried out in duplicate at a rate of 350,000 mf/tube for each technique: phenol/chloroform, commercial Qiagen kit, salting out, Tris-EDTA, methanol, and cetyltrimethylammonium bromide (CTAB). The integrity, purity, concentration, and quality of the DNA extracts were successively verified by agarose gel electrophoresis, spectrophotometry (A260/A280 and A260/A230 wavelength ratio), Qubit fluorometry, and endonuclease and polymerase activity. The six techniques were compared on the basis of the following parameters: concentration, purity, efficiency, effectiveness, integrity, safety of the technique, as well as cost and duration of the protocol. RESULTS: The ratios of the optical densities of the extracts A260/A280 and A260/A230 were, respectively: phenol/chloroform (1.82; 1.11), Qiagen (1.93; 1.36), salting-out (1.9; 2.04), Tris-EDTA (1.99; 1.183), methanol (2.126; 1.343), and CTAB (2.01; 2.426). The DNA yield was: phenol/chloroform (3.920 µg), Qiagen (10.280 µg), salting-out (10.390 µg), Tris-EDTA (0.5528 µg), methanol (0.1036 µg), and CTAB (1.115 µg). Endonuclease and polymerase activity was demonstrated by digestion of DNA and through amplicons obtained via polymerase chain reaction assays with phenol/chloroform, Qiagen, and salting-out extracts. CONCLUSION: The phenol/chloroform, Qiagen, and salting-out DNA extracts were all of good quality. Salting out had the best yield followed by Qiagen and then phenol/chloroform. Endonuclease and polymerase activity was effective in all three extracts despite the presence of some contaminants. These methods are therefore suitable for the extraction of DNA from Loa loa microfilariae. Tris-EDTA and methanol did not show adequate sensitivity, while CTAB was found to be unsuitable.

Loa loa infection detection using biomarkers: current perspectives.

Loa loa is originally a restricted filarial worm from central Africa and some west African countries. However, numerous imported cases are being reported throughout the world due to human movement. Traditionally, its diagnosis is based on identification of microfilariae in the peripheral blood or the passage of the adult worm under the conjunctiva. However, few patients have microfilariae in their peripheral blood, while the majority of infected people are amicrofilaremic (without microfilariae in their blood), despite clinical symptoms suggesting L. loa infection. This situation suggests that diagnoses based on the presence of microfilariae in the blood or the ocular passage of an adult worm, are not sensitive. Therefore, it seems necessary to search for biomarkers to remedy this situation. Furthermore, L. loa is a major obstacle in the control of other filarial worms in areas where these filariae are co-endemic. To develop a diagnostic tool based on a biomarker, several approaches have been considered using antibodies, antigens or nucleic acid detection. However, none of the diagnostic techniques in loiasis based on biomarkers has reached the point of care as have microscopic detection of microfilariae or observation of ocular passage of a worm.