Clinical epidemiology and high genetic diversity amongst Cryptococcus spp. isolates infecting people living with HIV in Kinshasa, Democratic Republic of Congo.

Neuromeningeal cryptococcosis (NMC) is a life-threatening opportunistic infection in advanced HIV disease patients (AHDP). It is caused by Cryptococcus spp. complexes and mainly occurs in sub-Saharan Africa. In this study, we performed molecular characterization and antifungal susceptibility profiling of Cryptococcus isolates from AHDP in Kinshasa (DRC). Additionally, we investigated a possible association between NMC severity factors and the Cryptococcus neoformans (Cn) multilocus sequence typing (MLST) profiles. We characterized the isolates using PCR serotyping, MALDI-TOF MS, internal transcribed spacer (ITS) sequencing, and MLST. Susceptibility testing for the major antifungal drugs was performed according to the EUCAST guidelines. Parameters associated with NMC severity, such as hypoglycorrhachia (< 50 mg/dL), increased cerebral spinal fluid opening pressure (> 30 cm H2O), and poor therapeutic outcome were compared with the Cn MLST sequences type (ST). Twenty-three out of 29 Cryptococcus isolates were identified as serotype A using PCR serotyping (79.3%; 95% IC: 65.5-93.1), while six (20.7%; 95% IC: 6.9-34.5) were not serotypable. The 29 isolates were identified by ITS sequencing as follows: Cryptococcus neoformans (23/29, 79.3%), Cutaneotrichosporon curvatus (previously called Cryptococcus curvatus) (5/29, 17.2%), and Papiliotrema laurentii (Cryptococcus laurentii) (1/29, 3.5%). Using the ISHAM MLST scheme, all Cn isolates were identified as molecular type VNI. These comprised seven different STs: ST93 (n = 15), ST5 (n = 2), ST53 (n = 1), ST31 (n = 1), ST4 (n = 1), ST69 (n = 1), and one novel ST that has not yet been reported from other parts of the world and was subsequently assigned as ST659 (n = 2). Of the included strains, only Papiliotrema laurentii was resistant to amphoterin B (1/29, 3.5%), 6.8% (2/29) were resistant to 5-flucytosine (the single Papiliotrema laurentii strain and one Cryptococcus neoformans isolate), and 13.8% (4/29) to fluconazole, including two of five (40%) Cutaneotrichosporon curvatus and two of 23 (8.7%) C. neoformans strains. We found a significative association between poor therapeutic outcome and a non-ST93 sequence type of causative strains (these concerned the less common sequence types: ST53, ST31, ST5, ST4, ST659, and ST69) (87.5% versus 40%, p = 0.02). Molecular analysis of Cryptococcus spp. isolates showed a wide species diversity and genetic heterogenicity of Cn within the VNI molecular type. Furthermore, it is worrying that among included strains we found resistances to several of the commonly used antifungals.

The seroprevalence of parvovirus B19 antibody in blood donors at the National Blood Transfusion Center in Kinshasa.

BACKGROUND: Human parvovirus B19 (PVB19) is a cosmopolitan DNA virus transmissible parenterally by blood transfusion. Therefore, the risk of transmission through asymptomatic blood donors should be considered and appropriately managed worldwide. PVB19 screening of blood and blood products for transfusion is not done routinely in the Democratic Republic of Congo (DRC). The main objective of this study was to determine the seroprevalence of PVB19 infection in healthy eligible blood donors in Kinshasa, capital of the DRC, located in the western part of the DRC, and the association of infection with the sociodemographic characteristics of blood donors. MATERIALS AND METHODS: A total of 360 whole blood donors who attended the National Center of Blood Transfusion were examined for anti-PVB19 IgG and IgM antibodies by using enzyme-linked immunosorbent assay kits. Sociodemographic information was collected on the blood donors. All statistical analyses were performed with SPSS 21. RESULTS: Among the study group, 289 men and 52 women were infected with PVB19. The mean age was 32.7 ± 9.8 years, 48.6% of donors were positive only for PVB19 IgG antibodies while 40.8% were positive for both IgG and IgM antibodies. In addition, 5.3% were positive only for PVB19 IgM antibodies and so were considered as a potential group of PVB19 transfusion-transmission. PVB19 seropositivity was significantly associated with sex, with a higher prevalence in men. In multivariate analysis, male sex and Tshangu district have emerged as major factors associated to PVB19 seropositivity. CONCLUSIONS: This research showed that recipients of blood and blood products in Kinshasa are at a high risk (5.3%) of transfusion-transmitted PVB19 infection. Therefore, the implementation of PVB19 nucleic acid testing assays capable of detecting all PVB19 genotypes and discard donations with high titer PVB19 DNA for blood products seems to be necessary.