Efficiency of Real-Time PCR in the Diagnosis of Community-Acquired Bacterial Meningitis in Children

S. Raza, Bimal Das, R. Chaudhry, V. Goyal, R. Lodha, S. Sood, H. Gautam, A. Kapil
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Abstract

Objectives: Community-acquired bacterial meningitis (CABM) is a life-threatening condition and remains a public health concern despite various efforts to prevent it. This study aimed to detect the bacteria causing CABM in children by Real-Time PCR. Methods: In total, 178 Cerebrospinal fluid (CSF) samples from suspected meningitis cases were collected and subjected to cell count, biochemical, microbiological, and molecular analysis. Bacteria grown on blood and chocolate agar were identified by matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS). DNA from CSF was extracted and used to detect bacteria by Real-Time PCR using TaqMan Probe. Results: Fifty (28.09%) patients were diagnosed with confirmed meningitis. Of them, 46 (25.84%) were Real-Time PCR, and four (2.25%) were culture and Real-Time PCR positive. Out of 50 bacteria detected, S. pneumoniae (n=35, 19.7%) was the leading causative bacteria and was followed by H. influenzae (seven, 3.93%), E. coli (five, 2.80%), S. agalactiae (two, 1.12%), and N. meningitidis (one, 0.56%). Most of the S. pneumoniae (18 isolates, 51.4%) were isolated from 3-24 months of children, and in neonates, E. coli was the predominant bacteria. When CSF culture was the gold standard for diagnosis, the sensitivity and specificity of Real-Time PCR for S. pneumoniae were 100% (95%CI: 15.81-100%) and 81.25% (95%CI: 74.69-86.73%), respectively. Conclusion: Streptococcus pneumoniae remains the leading organism of CABM in children despite vaccination and advancement in diagnosis. Real-time PCR has emerged as a vibrant diagnostic molecular appliance. Hence, Regular surveillance is crucial to curb the burdens and trends of CABM in children.
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实时荧光定量PCR对儿童社区获得性细菌性脑膜炎的诊断价值
目的:社区获得性细菌性脑膜炎(CABM)是一种危及生命的疾病,尽管采取了各种预防措施,但仍是一个公共卫生问题。本研究旨在利用Real-Time PCR检测儿童CABM病原菌。方法:收集疑似脑膜炎患者脑脊液178份,进行细胞计数、生化、微生物学及分子分析。采用基质辅助激光解吸电离飞行时间质谱法(MALDI-TOF MS)对生长在血液和巧克力琼脂上的细菌进行了鉴定。提取脑脊液DNA,采用TaqMan探针进行实时荧光定量PCR检测细菌。结果:确诊脑膜炎50例(28.09%)。其中,Real-Time PCR阳性46例(25.84%),培养和Real-Time PCR阳性4例(2.25%)。50种病原菌中,肺炎链球菌35种,占19.7%,其次是流感嗜血杆菌7种,占3.93%,大肠杆菌5种,占2.80%,无乳链球菌2种,占1.12%,脑膜炎奈瑟菌1种,占0.56%。大多数肺炎链球菌(18株,51.4%)来自3 ~ 24月龄儿童,在新生儿中以大肠杆菌为优势菌。当CSF培养为诊断金标准时,Real-Time PCR对肺炎链球菌的敏感性为100% (95%CI: 15.81 ~ 100%),特异性为81.25% (95%CI: 74.69 ~ 86.73%)。结论:尽管疫苗接种和诊断的进步,肺炎链球菌仍然是儿童CABM的主要病原体。实时聚合酶链反应已成为一个充满活力的诊断分子设备。因此,定期监测对于遏制儿童CABM的负担和趋势至关重要。
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