Variation of prevalence of malaria, parasite density and the multiplicity of Plasmodium falciparum infection throughout the year at three different health centers in Brazzaville, Republic of Congo.

BACKGROUND: In the Republic of Congo, hot temperature and seasons distortions observed may impact the development of malaria parasites. We investigate the variation of malaria cases, parasite density and the multiplicity of Plasmodium falciparum infection throughout the year in Brazzaville. METHODS: From May 2015 to May 2016, suspected patients with uncomplicated malaria were enrolled at the Hôpital de Mfilou, CSI « Maman Mboualé¼, and the Laboratoire National de Santé Publique. For each patient, thick blood was examined and parasite density was calculated. After DNA isolation, MSP1 and MSP2 genes were genotyped. RESULTS: A total of 416, 259 and 131 patients with suspected malaria were enrolled at the CSI «Maman Mboualé¼, Hôpital de Mfilou and the Laboratoire National de Santé Publique respectively. Proportion of malaria cases and geometric mean parasite density were higher at the CSI «Maman Mboualé¼ compared to over sites (P-value <0.001). However the multiplicity of infection was higher at the Hôpital de Mfilou (P-value <0.001). At the Laboratoire National de Santé Publique, malaria cases and multiplicity of infection were not influenced by different seasons. However, variation of the mean parasite density was statistically significant (P-value <0.01). Higher proportions of malaria cases were found at the end of main rainy season either the beginning of the main dry season at the Hôpital de Mfilou and the CSI «Maman Mboualé¼; while, lowest proportions were observed in September and January and in September and March respectively. Higher mean parasite densities were found at the end of rainy seasons with persistence at the beginning of dry seasons. The lowest mean parasite densities were found during dry seasons, with persistence at the beginning of rainy seasons. Fluctuation of the multiplicity of infection throughout the year was observed without significance between seasons. CONCLUSION: The current study suggests that malaria transmission is still variable between the north and south parts of Brazzaville. Seasonal fluctuations of malaria cases and mean parasite densities were observed with some extension to different seasons. Thus, both meteorological and entomological studies are needed to update the season's periods as well as malaria transmission intensity in Brazzaville.

No polymorphisms in K13-propeller gene associated with artemisinin resistance in Plasmodium falciparum isolated from Brazzaville, Republic of Congo.

BACKGROUND: In the Republic of Congo, artemisinin-based combinations have been recommended for the treatment of uncomplicated malaria since 2006. However, the emergence of resistant parasites again these combinations in Southeast Asia is a threat for the control of this disease, especially in sub-Saharan Africa where the weight of the disease is important. Indeed, polymorphisms in Plasmodium falciparum K13-propeller gene have been involved in variations of drug sensitivity of Plasmodium falciparum to artemisinin-based combinations. The aim of the current study is to determine the prevalence of mutations of this gene in isolates collected in three health centers in Brazzaville. METHODS: From May 2015 to May 2016, a total of 131, 259 and 416 samples from patients with suspected malaria were collected at the Laboratoire National de Santé Publique, Hôpital de Mfilou, and the CSI «Maman Mboualé¼ respectively. After DNA isolation, genotyping and sequencing of Plasmodium falciparum K13-propeller were performed in positive Plasmodium falciparum isolates identified after msp-2 gene genotyping. RESULTS: All 806 samples collected were msp-2 genotyped and Plasmodium falciparum infections were confirmed in 287 samples with 43, 85, 159 samples from Laboratoire National de Santé Publique, Hôpital de Mfilou, and the CSI «Maman Mboualé¼ respectively. Of these 287 msp-2 positives samples, K13-propeller nested PCR products were successfully obtained from 145 (50.52%) isolates and sequences were generated from 127(87.58%) nested products. None of mutations that were associated with ACTs resistance in Southeast Asia were detected on the samples from three different study sites from Brazzaville. However, one mutation type was observed at position 578, where alanine was substituted by serine (A578S) in two isolates (1.57%, 2/127), those from the Hôpital de Mfilou. No mutation was found in isolates from the two other sites. CONCLUSION: The current study shows a very limited polymorphism in the K13-propeller gene in isolates from the Republic of Congo and K13 polymorphisms associate with ACT resistance are not present in this country. However, permanent and large surveillance of resistant parasite population using K13-propeller gene is recommended.

Evaluation of Routine Microscopy Performance for Malaria Diagnosis at Three Different Health Centers in Brazzaville, Republic of Congo.

BACKGROUND: In Republic of Congo, malaria diagnosis still widely relies on microscopy. We aimed to evaluate the performance of routine microscopy for malaria diagnosis at three different health centers in Brazzaville. METHODS: A total of 259, 416, and 131 patients with clinical signs of uncomplicated malaria were enrolled at the Hôpital de Mfilou, Centre de Santé Intégré "Maman Mboualé," and Laboratoire National de Santé Publique, respectively. Two thick blood smears were prepared for each patient, the first being examined by routine microscopists and the second by expert. RESULTS: At the Hôpital de Mfilou, sensitivity was 62.1% and specificity was 67.3%. Positive and negative predictive values were 55.6% and 72.9%, respectively. At the Centre de Santé Intégré "Maman Mboualé," sensitivity was 94.2% and specificity was 33.6%. Positive and negative predictive values were 50% and 89.1%, respectively. At the Laboratoire National de Santé Publique, sensitivity and specificity were high with 91.7% and 94.9%, respectively. Positive and negative predictive values were 64.7% and 99.1%, respectively. CONCLUSION: The performance of routine malaria microscopy in Brazzaville remains inaccurate with large variations among different health centers. Therefore, repeated training including supervision and evaluation would improve routine malaria diagnosis for better management of malaria in Brazzaville, the Republic of Congo.