Early Diagnosis of Viral Pathogens in Cerebrospinal Fluid from Patients with Acute Meningitis

• Main Authors: Candan ÇİÇEK, Hüsnü PULLUKÇU, Hale KALFAOĞLU, Hasip KAHRAMAN, Eylem ULAŞ SAZ
• Format: Article
• Language: English
• Published: Bilimsel Tip Yayinevi 2015-12-01
• Series: Flora Infeksiyon Hastalıkları ve Klinik Mikrobiyoloji Dergisi
• Subjects: Acute meningitis; Multiplex PCR; Shell-vial cell culture; Virus
• Online Access: http://www.floradergisi.org/getFileContent.aspx?op=REDPDF&file_name=2015-20-04-174-181.pdf

Introduction: Viruses are the major causes of acute meningitis and encephalitis. The aim of this study was to establish the prevalence of viruses in patients with acute meningitis. Materials and Methods: Cerebrospinal fl uid specimens were collected from 111 patients [41 (37%) female, 70 (63%) male] with central nervous system infections between April 2012 and February 2014. The age range of patients was between 1 to 88 years (median: 35.5 years). Of the 111 patients, 22 (19.8%) were pediatric patients (median: 5), and 89 (80.2%) were adult patients (median: 37). Multiplex polymerase chain reaction (PCR) assay [herpes simplex virus (HSV) 1-2, Varicella zoster virus (VZV), human herpes virus (HHV)-6, Epstein-Barr virus (EBV), cytomegalovirus (CMV) enterovirus)] (Seegene Inc., Seoul Korea) and real time PCR [West Nile virus (WNV)] (Lanciotti RS, 2000) methods were performed on all clinical specimens. In addition, to the 39 specimens which had suffi cient amount of CSF; shell-vial cell culture method was performed simultaneously with multiplex PCR. In order to isolate HSV 1-2 and enteroviruses; Vero cell line and fl uorescein isothiocynate labelled each virus specifi c polyclonal antibodies were used (HSV 1-2, Panenetrovirus, Light Diagnostic, Millipore, ABD). After nucleic acid extraction and complementary DNA (cDNA) synthesis from clinical specimens, multiplex amplifi cation of nucleic acids were done by Dual Priming Oligonucleotide (DPO) primers and ‘‘Seeplex Meningitides V1-2 ACE Detection’’ kit (Seegene Inc., South Korea). Results: Out of 111 patients, 36 (32.4%) were positive and 75 (67.6%) were negative for HSV 1-2, VZV, HHV-6, EBV, CMV, enterovirus and WNV. Of the positive patients, 30 were adults (median: 39 years) and 6 (median: 7.5 years) were pediatric patients (p= 0.564). Of the positive specimens, 25 (69.4%) were enterovirus (pediatric: 6, adult: 19), (single: 20, multiple: 5), 11 (30.5%) were herpes viruses (pediatric: 1, adult: 10), (single: 6, multiple: 5), 9 (25%) were WNV (pediatric: 1, adult: 8), (single: 3, multiple: 6). CMV was not identifi ed in any patient. Multiple viruses were identifi ed in 8 (7.2%) of the patients (pediatric: 1, adult: 7). We didn’t detect HSV 1-2 in any of the patients; however, in only one patient, enterovirus was detected with shell vial cell culture method. Conclusion: Viruses were detected in approximately 33% of the patients with acute meningitis. There was no statistically signifi cant difference between positivity rates in adults and children. Most commonly detected viruses were enteroviruses (22.5%) in children and adults. This rate was followed by herpes viruses approximately at the percentage of 10%. Herpes viruses were most frequently identified in adults. Multiple virus infections were detected in 7.2% of patients and it was observed that the majority of these were in the adult group. The WNV was often accompanied with multiple viral infections and was seen in the majority of the adult group. CMV was not identified in any patient. Shell-vial cell culture methods were compatible with the multiplex PCR results.