Bhagchandani Sharda P, Kubade Sushant, Nikhare Priyanka P, Manke Sonali, Chandak Nitin H, Kabra Dinesh, Baheti Neeraj N, Agrawal Vijay S, Sarda Pankaj, Mahajan Parikshit, Ganjre Ashish, Purohit Hemant J, Singh Lokendra, Taori Girdhar M, Daginawala Hatim F, Kashyap Rajpal S
Research Laboratory, Central India Institute of Medical Sciences, Nagpur, Maharashtra, India.
Environmental Genomics Unit, National Environmental Engineering Research Institute, Nagpur, Maharashtra, India.
Mol Diagn Ther. 2016 Feb;20(1):45-54. doi: 10.1007/s40291-015-0174-z.
Bacterial meningitis is a dreadful infectious disease with a high mortality and morbidity if remained undiagnosed. Traditional diagnostic methods for bacterial meningitis pose a challenge in accurate identification of pathogen, making prognosis difficult. The present study is therefore aimed to design and evaluate a specific and sensitive nested 16S rDNA genus-based polymerase chain reaction (PCR) assay using clinical cerebrospinal fluid (CSF) for rapid diagnosis of eight pathogens causing the disease.
The present work was dedicated to development of an in-house genus specific 16S rDNA nested PCR covering pathogens of eight genera responsible for causing bacterial meningitis using newly designed as well as literature based primers for respective genus. A total 150 suspected meningitis CSF obtained from the patients admitted to Central India Institute of Medical Sciences (CIIMS), India during the period from August 2011 to May 2014, were used to evaluate clinical sensitivity and clinical specificity of optimized PCR assays.
The analytical sensitivity and specificity of our newly designed genus-specific 16S rDNA PCR were found to be ≥92%. With such a high sensitivity and specificity, our in-house nested PCR was able to give 100% sensitivity in clinically confirmed positive cases and 100% specificity in clinically confirmed negative cases indicating its applicability in clinical diagnosis.
Our in-house nested PCR system therefore can diagnose the accurate pathogen causing bacterial meningitis and therefore be useful in selecting a specific treatment line to minimize morbidity. Results are obtained within 24 h and high sensitivity makes this nested PCR assay a rapid and accurate diagnostic tool compared to traditional culture-based methods.
细菌性脑膜炎是一种严重的传染病，如果未被诊断出来，死亡率和发病率都很高。细菌性脑膜炎的传统诊断方法在准确鉴定病原体方面存在挑战，难以进行预后判断。因此，本研究旨在设计并评估一种基于16S rDNA属特异性的巢式聚合酶链反应（PCR）检测方法，该方法使用临床脑脊液（CSF）快速诊断导致该疾病的8种病原体。
本研究致力于开发一种内部属特异性16S rDNA巢式PCR，该方法涵盖了导致细菌性脑膜炎的8个属的病原体，使用新设计的以及基于文献的各属引物。从2011年8月至2014年5月期间入住印度中央印度医学科学研究所（CIIMS）的患者中获得的总共150份疑似脑膜炎脑脊液样本，用于评估优化后的PCR检测方法的临床敏感性和临床特异性。
我们新设计的属特异性16S rDNA PCR的分析敏感性和特异性均≥92%。凭借如此高的敏感性和特异性，我们的内部巢式PCR在临床确诊的阳性病例中能够达到100%的敏感性，在临床确诊的阴性病例中能够达到100%的特异性，表明其在临床诊断中的适用性。
因此，我们的内部巢式PCR系统能够诊断出导致细菌性脑膜炎的准确病原体，从而有助于选择特定的治疗方案以降低发病率。该方法在24小时内即可获得结果，与传统的基于培养的方法相比，高敏感性使这种巢式PCR检测成为一种快速、准确的诊断工具。
