Sharma Nupur, Gautam Hitender, Tyagi Sonu, Raza Shahid, Mohapatra Sarita, Sood Seema, Dhawan Benu, Kapil Arti, Das Bimal K
Department of Microbiology, All India Institute of Medical Sciences, New Delhi, India.
J Family Med Prim Care. 2022 Feb;11(2):593-598. doi: 10.4103/jfmpc.jfmpc_1162_21. Epub 2022 Feb 16.
Prompt and accurate diagnosis of acute bacterial meningitis (ABM) is critical for patient management. We designed and evaluated two sets of multiplex-PCR assays for the simultaneous detection of six major etiologies of ABM i.e., type b, and in one set and in another set of multiplex-PCR in CSF of patients with suspected ABM.
A total of 113 CSF specimens from patients of all ages having clinical features suggestive of meningitis were tested for bacteriological evidence by Gram's smear, culture, and our designed multiplex-PCR.
Multiplex-PCR assay performed excellently by increasing the overall detection rate by 6% when compared to culture as of total 113 samples tested, 17 (15%) were positive by multiplex-PCR whereas only 9% (10/113) were positive by culture. It detected the DNA in eight culture negative samples revealing the presence of in three and other possible bacterial pathogens in five of them. Our assay showed a DNA detection limit of 1 pg/μL. Compared to CSF culture, the sensitivity and specificity of the multiplex-PCR were 90% and 92.2%, respectively.
This study accentuates the importance of multiplex-PCR assay that is efficiently fast and reliable for the diagnosis of acute bacterial meningitis that can substantially improve the diagnosis in culture negative cases, especially in patients who were previously started on antimicrobial therapy.
急性细菌性脑膜炎（ABM）的及时准确诊断对患者管理至关重要。我们设计并评估了两组多重聚合酶链反应（PCR）检测方法，用于同时检测ABM的六种主要病因，即在一组多重PCR中检测b型[细菌名称未给出]，在另一组多重PCR中检测[另一种细菌名称未给出]，用于疑似ABM患者脑脊液检测。
对113例具有脑膜炎临床特征的各年龄段患者的脑脊液标本进行革兰氏涂片、培养及我们设计的多重PCR检测，以寻找细菌学证据。
在总共检测的113个样本中，多重PCR检测表现出色，与培养相比，总体检测率提高了6%。多重PCR检测出17例（15%）阳性，而培养仅检测出9%（10/113）阳性。它在8个培养阴性样本中检测到了DNA，其中3个样本检测到[细菌名称未给出]，另外5个样本检测到其他可能的细菌病原体。我们的检测方法显示DNA检测限为1 pg/μL。与脑脊液培养相比，多重PCR的敏感性和特异性分别为90%和92.2%。
本研究强调了多重PCR检测在急性细菌性脑膜炎诊断中的重要性，该检测方法高效、快速且可靠，可显著改善培养阴性病例的诊断，特别是对于那些先前已接受抗菌治疗的患者。
