Browsing by Author "Lema Ayele"

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    Comparison of the Efficiency of CSF and Serum PCR in the Diagnosis of Bacterial Meningitis Among Suspected Cases at Tikur Anbessa Specialized Hospital and Yekatit 12 Hospital Medical College, Addis Ababa Ethiopia
    (Addis Ababa University, 2025-02-27) Lema Ayele; Daniel Asrat; Getachew Tesfaye
    Background Early and accurate diagnosis of bacterial meningitis reduces mortality and sequelae. In resource-limited settings like Ethiopia, where laboratory diagnostics are scarce, developing a diagnostic tool based on an easy-to-collect blood sample for the diagnosis of bacterial meningitis is crucial to improving the diagnosis and prompt treatment initiation that has a great impact on patient outcomes. Objective To compare the efficiency of cerebrospinal fluid and blood PCR for the diagnosis of bacterial meningitis in patients suspected of meningitis. Methods An institutional-based cross-sectional study was conducted at Tikur Anbessa Specialized Hospital and Yekatit 12 Hospital Medical College from August 2023 to February 2024. A total of 202 patients who were suspected of having meningitis were given their consent to collect the demographic, clinical data, CSF, and blood samples. Demographic data and clinical samples (CSF and Blood) were collected by using pre-tested standardized questionnaires and aseptic techniques, respectively. Conventional laboratory analysis and bacterial isolation using aliquots of CSF were performed following the standard microbiology laboratory procedures. The leftover CSF and serum samples were stored at -20°C until transported to the Armauer Hansen Research Inistitute (AHRI) for further molecular tests. Genomic DNA from CSF and serum was extracted by the Bioer NPA-32P automated nucleic acid purification machine. A multiplex PCR technique was used to detect the presence of Neisseria meningitidis, Haemophilus influenzae, Streptococcus pneumoniae, Streptococcus agalactiae, Escherichia coli, Staphylococcus aureus, Listeria monocytogenes, and Klebsiella pneumoniae. The end product of the PCR amplicon was observed under the Vilber machine after staining with ethidium bromide. Data were entered into Epi Info 7, cleaned, exported to Excel, and loaded into SPSS version 26 software for analysis. Descriptive statistics were presented in tables and bar charts. The agreement between the tests to detect bacterial meningitis was calculated by using an SPSS contingency table. Result Out of 202 study participants, 54% (n = 109) were males, and 88.6% (n = 179) of them were from the pediatric population, with an average age of 1.45 ± 0.69 years. Reduced ability to breastfeed was the most common clinical presentation 74.8% (n = 151), followed by high fever 71.3% (n = 144) and a loss of consciousness 66.3% (n = 134). Of 202 CSF, 5.4% (n = 11) and 4.5% (n = 9) of the samples tested were positive for gram stain and culture, respectively. Overall, bacterial DNA was detected in 26% (n = 54) and 46% (n = 93) CSF and serum samples, respectively. The concordance between CSF and serum PCR was observed in 34 (17%) cases. E. coli was the most predominant bacteria detected in both CSF and serum PCR at 55.5% (n = 30) and 61.2% (n = 57), respectively, followed by S. pneumoniae detected in both CSF and serum PCR at 15% (n = 8) and 29% (n = 27), respectively. The overall agreement between CSF and serum PCR is 60.9% with a positive percent agreement, and negative percent agreement between the two tests is 62.9% and 60.1%, respectively. Conclusion Bacterial DNA was 20% more frequently detected in serum compared to CSF, and 42% compared to culture in suspected cases of meningitis. Furthermore, these findings are clinically very important to minimise the invasive lumbar puncture for CSF collection. Overall, serum PCR could be considered as a supplementary diagnostic tool, a less invasive diagnostic procedure for the diagnosis of bacterial meningitis. However, further similar studies are important using qPCR and both blood and CSF culture.