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印度中部人群中通过定量实时聚合酶链反应确定病毒性中枢神经系统感染的病因和流行病学

Determination of Etiology and Epidemiology of Viral Central Nervous System Infections by Quantitative Real-Time Polymerase Chain Reaction in Central India Population.

作者信息

Tiwari Ragini I, Bhullar Shradha S, Chandak Nitin H, Baheti Neeraj N, Daginawala Hatim F, Singh Lokendra R, Kashyap Rajpal S

机构信息

Biochemistry Research Laboratory, Central India Institute of Medical Sciences, Nagpur, Maharashtra, India.

出版信息

Neurol India. 2020 Nov-Dec;68(6):1323-1330. doi: 10.4103/0028-3886.304080.

DOI:10.4103/0028-3886.304080
PMID:33342862
Abstract

INTRODUCTION

Viral infections of the central nervous system (CNS) are the most common cause of hospital admission in worldwide and remain a challenging disease for diagnosis and treatment. The most common infectious agents associated with viral CNS infections are cytomegalovirus (CMV), Epstein-Barr virus (EBV), varicella zoster virus (VZV), Japanese encephalitis virus (JEV), Dengue virus (DENV),West Nile virus(WNV), and Chandipura virus(CHPV). The aim of the present work was to find the etiology of CNS viral infection in the Central India population by transcriptase PCR (RT-PCR) comparing real-time polymerase chain reaction (PCR) method [one-step and two-step reverse transcriptase (RT-PCR)] in cerebrospinal fluid (CSF) and blood samples of CNS viral infections patients.

MATERIALS AND METHODS

One-step and two-step real-time PCR assays were evaluated in CSF and parallel blood samples from patients with viral CNS infections for detection of DNA and RNA viruses. A comparative analysis was also done between gDNA, gRNA, cDNA, and plasmid-based real-time PCR methods for an efficient quantitation of viral particles in clinical samples for determination of viral etiology.

RESULT

On evaluation of 150 CSF and 50 parallel blood samples from suspected cases of viral CNS infections, a viral etiology was confirmed in 21 (14%) cases, including 3% for EBV, 1% of CMV, and 5% for VZV and JEV. The one-step RT-PCR has a higher detection limit for detection and quantitation of viral RNA in comparison to two-step RT-PCR.

CONCLUSION

Our result reveals that VZV and JEV are the most usual cases of CNS viral infection in hospitalized patients in the Central India population and one-step RT-PCR shows higher viral load detection limits for quantitation of viral genome and more sensitivity in comparison to two-step RT-PCR.

摘要

引言

中枢神经系统(CNS)病毒感染是全球范围内医院收治患者的最常见病因,并且在诊断和治疗方面仍然是一种具有挑战性的疾病。与病毒性中枢神经系统感染相关的最常见感染因子是巨细胞病毒(CMV)、爱泼斯坦-巴尔病毒(EBV)、水痘带状疱疹病毒(VZV)、日本脑炎病毒(JEV)、登革病毒(DENV)、西尼罗河病毒(WNV)和钱迪普拉病毒(CHPV)。本研究的目的是通过转录酶聚合酶链反应(RT-PCR),比较实时聚合酶链反应(PCR)方法[一步法和两步法逆转录酶(RT-PCR)],来确定印度中部人群中枢神经系统病毒感染的病因,样本来自中枢神经系统病毒感染患者的脑脊液(CSF)和血液。

材料与方法

对病毒性中枢神经系统感染患者的脑脊液和并行血液样本进行一步法和两步法实时PCR检测,以检测DNA和RNA病毒。还对基于基因组DNA(gDNA)、基因组RNA(gRNA)、互补DNA(cDNA)和质粒的实时PCR方法进行了比较分析,以便有效定量临床样本中的病毒颗粒,从而确定病毒病因。

结果

对150份脑脊液样本和50份来自疑似病毒性中枢神经系统感染病例的并行血液样本进行评估后,在21例(14%)病例中确认了病毒病因,其中EBV占3%,CMV占1%,VZV和JEV各占5%。与两步法RT-PCR相比,一步法RT-PCR在检测和定量病毒RNA方面具有更高的检测限。

结论

我们的结果表明,VZV和JEV是印度中部人群住院患者中枢神经系统病毒感染最常见的病例,与两步法RT-PCR相比,一步法RT-PCR在定量病毒基因组时显示出更高的病毒载量检测限和更高的灵敏度。

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