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دراسة معظم السمات المناعية وتحديد البكتريا الابرز في مرض التهاب السحايا باستعمال تقنية البي سي ار == Study Some Immunological Aspects And Determination Of Most Prominent Bacteria In Meningitis Patients By Using Real Time Pcr
Author name:
شيماء عبد الرضا حميد
Supervisor name:
زينب فاضل عاشور | دريد قاسم الشريف
General topic:
Medicine
Specific topic:
Microbiology - Immunology
Degree:
Master
University:
Mustansiriyah University
Language:
English
University location:
Baghdad
First pages:
19T1142 - p.pdf
Abstract:
Meningitis, which is inflammation of the membranes that surround the brain and spinal cord and can occur at any age, it is caused by infectious microorganisms include (Bacteria, Viruses, Fungi, and Parasites) and non infectious include (Drugs, Carcinoma and Inflammatory disorder), also requires early treatment with aggressive use of potent antimicrobials; otherwise the outcome almost ends fatally.The predominant causes of bacterial meningitis are streptococcus pneumoniae, Haemophilus influenzae type b (Hib) and Neisseria meningitides.This prospective study includes 40 cases of meningitis which have been admitted to the Baghdad Teaching Hospital and Child Welfare Teaching Hospital during the period from May to November/ 2014.The detection of meningitis causative agent were specified by using the standard biochemical and microbiological methods such as : Determination of protein and sugar in Cerebrospinal fluid, Differential and cell count, growth on Culture media and morphological appearance on Gram's stain.Confirmative tests were also done for identification of meningitis causative agent; these tests included Procalcitonin, IL - 6, CRP, D. Dimer. These confirmative tests have been crowned by PCR technique, by employing the detection of the bacterial gene.The DNA of bacteria was extracted from CSF of meningitis patients and detection the type of bacteria gene by using multiplex PCR technique that using primers to amplify each ply, bexA and ctrA gene.Out of 40 samples, 28 (70%) were identified as bacterial meningitis and 12 (30%) as viral meningitis, the result of PCR reaction revealed unmistakably that 20 (71.4%) of 28 were detected the bacteria (S. pneumonia, H. influenzaetype b and N. meningitides) as a causative agent, while there were no detectable infectious agents in the (40) control group included in this study.Viral meningitis patients showed the number of lymphocytes (31.92 cell/ mm3) and neutrophils (1.00 cell/ mm3), while the highest number of lymphocytes and neutrophils in bacterial meningitis caused by S. pneumoniae (77.08 cell/ mm3) , (28.86 cell/ mm3) respectively, so that there was significant difference between cells (lymphocyte and neutrophil) with study group at P value = 0.010, P value = 0.013 respectively. In microbiological examination 22.5% of patients group (40) showed positive result of culture and 7.5% showed positive result of gram stain.In biochemical tests, the detection of Cerebrospinal fluid's protein and glucose level was done. The level of protein concentrations in bacterial meningitis with S. pneumoniae (103.08 mg/l), H. influenzae (65.14 mg/l) and in other bacteria was (124.44 mg/l) which is higher than the protein concentration in viral meningitis (42.25 mg/l), and these results showed highly significant difference at (P=0.010). Results of glucose's level determination showed that there were no significant difference among the studied groups (P =0.2). Results of glucose's level determination showed the presence of non significant difference among the studied groups, (60.92 mg/dl), (43, 57 mg/ dl), (51.00 mg/dl), (41.50 mg/dl) consecutively appeared in bacterial, viral.Interleukin - 6 and Procalcitonin levels were diagnosed by Enzyme Linked Immunosorbent Assay, while the plasma D. Dimer and erum C - Reactive protein was diagnosed by NaycoCard technique.The current results showed the presence of high concentration of IL - 6 among meningitis patients, high concentration of serum IL - 6 appeared in bacterial and viral meningitis (28.68 pg/ml) when compared with control group (2.194 pg/ml) (P =0.001), while CSF IL - 6 show high concentration in bothbacterial and viral meningitis but with no significant difference (579.24 pg/ml) (P =0.0001). The data of this study showed, there was correlation between IL - 6 levels and neutrophil in meningitis patients. Interleukin - 6 had significant correlation with neutrophil (P =0.009). While no significant correlation appeared between IL - 6 and Lymphocyte.Procalcitonin results showed high concentration in bacterial meningitis group compared with viral and control group with a highly significant differences (P =0.0001).The results of CRP shown no significant differences between bacterial and viral group, while the levels of D. Dimer appear significant differences in both groups when compared with control group.Current study indicated that PCR, Procalcitonin and D. Dimer played an important role in diagnosis of meningitis disease.
