RESUMO
Method: In a comparative experimental cross-sectional study, RNA was extracted from oral swabs and blood samples from 25 healthy individuals at the Department of Molecular Medicine, KNUST. RNA was extracted by the manual AGPC extraction method and commercial RNA extraction kits. The quantity (ng/µl) and purities (260/280 nm) of the extracted RNA were measured spectrophotometrically using the IMPLEN NanoPhotometer® N60. The presence of RNA in the extracts was confirmed using 2% agarose gel electrophoresis. Statistical analyses were conducted using R language. Results: The yield of RNA extracted from blood and oral swab samples using modified AGPC was significantly higher compared to the commercial methods (p < 0.0001). However, the purity of RNA extracted by the manual AGPC method from blood was significantly lower than the commercial methods (p < 0.0001). Moreover, the purity from oral swabs using the manual AGPC method was significantly lower compared to QIAamp (p < 0.0001) and the OxGEn kits method (p < 0.001). Conclusion: The modified manual AGPC method has a very high yield of RNA extracts using blood samples, which could serve as an alternate cost-effective method for RNA extraction in resource-limited laboratories; however, its purity may not be suitable for downstream processes. Moreover, the manual AGPC method may not be suitable for extracting RNA from oral swab samples. Future investigation is needed to improve the purity of the manual AGPC RNA extraction method and also confirmation of the obtained results by PCR amplification and RNA purity verification by sequencing.
Assuntos
Clorofórmio , Fenol , Humanos , Estudos Transversais , Laboratórios , Fenóis , RNARESUMO
Background: We aimed at investigating the impact of malaria on the haematological parameters of residents from different demographic settlements in the Ashanti Region of Ghana. Malaria parasites trigger changes in certain haematological parameters, which may result in a number of clinical manifestations. Differences in demographic settlements, such as rural, peri-urban and urban settlements may also influence these changes, but this has not been extensively studied in Ghana. Methods: We conducted a hospital-based, cross-sectional study from January to December 2018 in three different settlements. A total of 598 participants were recruited. Blood smears were examined to detect and quantify malaria parasitaemia, while haematological parameters were measured using a haematology analyser. Results: Participants from the rural settlement had the highest malaria prevalence (21.3%) compared to the urban (11.8%) and peri-urban areas (13.3%); however, the peri-urban area had the highest median parasite density (568; IQR=190.0-1312.0). Age was significantly associated with the odds of malaria positivity (OR: 0.97; CI:0.96 - 0.99). When haematological parameters of the malaria-infected study participants were compared to the parameters of uninfected participants, red blood cell count (p=0.017), haemoglobin (p=0.0165), haematocrit (p=0.0015), mean corpuscular volume (p=0.0014), plateletcrit (p<0.0001) and platelet count (p<0.0001) were all significantly lower in the malaria infected group. In addition to age, haemoglobin and plateletcrit levels were also inversely correlated with the odds of testing positive for malaria, suggesting that children who were anaemic and/or thrombocytopaenic were likely to be infected. After fitting the data to a logistic regression model comprising the three variables, the model correctly categorised 78% of uninfected study participants, but only 50% of the malaria-positive participants. Conclusions: Study participants who were positive for malaria were younger and had low haemoglobin and plateletcrit levels compared to uninfected individuals. Further studies are needed to more precisely elucidate the relationship between malaria infection,demographic and haematological parameters.
RESUMO
Background: Toxoplasma gondii is an obligate, intracellular, apicomplexan parasite that causes toxoplasmosis. Although the global prevalence of toxoplasmosis has been estimated to be approximately 30%, there is limited seroprevalence data in Ghana, with a dearth of information on the impact of T. gondii on haematological parameters in exposed persons. Methods: Questionnaires were administered to 300 consenting individuals to obtain demographic information and assessment of their risk of exposure to T. gondii. Using anti- T. gondii IgG/IgM combo test kits, seropositivity to parasite-specific IgG and/or IgM was determined. A haematological analyser was used to measure haematological parameters. Results: The participants included 58 males and 242 females, and ranged in age from 6 months to 84 years, with a median age of 27 years. There was an overall seroprevalence of 50.3% (n=151), with 49.7% (n=149) of the study participants seropositive for IgG and 1% (n=3) testing positive for IgM. Furthermore, the observed seroprevalence among pregnant women was 56.4% (n=62). With regards to the different communities in which the hospitals were located, a seroprevalence of 55.6% was observed in the rural community, 50.6% in the peri-urban community and 47.1% in the urban community. The study identified cat ownership, contact with cat litter [RR (95% CI: 1.76 (1.23-2.53), 1.66 (1.03-2.67), 1.25(1.00-1.57)] and age (p<0.001) as risk factors for infection. Analyses of haematological data also revealed significant differences between the red blood cell counts (p=0.038) and mean corpuscular volumes (p=0.0007) of seropositive and seronegative study participants. Conclusions: About half of the study population, including a significant number of women of reproductive age carried antibodies against T. gondii, raising questions about the risk of congenital toxoplasmosis, as well as possible links to anaemia. We, therefore, recommend that screening for Toxoplasma gondii be included in the routine screening of pregnant women seeking antenatal care.
