Molecular detection and genotyping of enteroviruses from CSF samples of patients with suspected sepsis-like illness and/or aseptic meningitis from 2012 to 2015 in West Bank, Palestine

Abstract

BackgroundHuman enteroviruses (HEVs) are the most frequently reported cause of aseptic meningitis with or without CSF pleocytosis in childhood. Rapid detection and genotype of HEVs is essential to determine the causative agent and variant causing sepsis-like illness and/or aseptic meningitis.AimTo investigate the molecular epidemiology of enteroviruses (EVs) among patients with sepsis-like illness and/or aseptic meningitis admitted to three major hospitals in West Bank, Palestine from 2012 to 2015.MethodsDuring the study period, 356 CSF samples were collected from patients with sepsis-like illness and/or aseptic meningitis. Two RT-nested PCR assays targeting a partial part of 5'UTR for direct diagnosis and the VP1 region for genotyping by sequence analysis of the viral genome were used.ResultsHEV RNA was detected in 66 of 356 (18.5%) of CSF samples. Age distribution showed that 64% (42/66) were infants (<1 year), 18% were children between 1 and 5 years old, 12% were children between 5 and 10 years old, and 6% were more than 10 years old. Of the 66 EV cases, 12 were successfully genotyped. Five different EV genotypes were identified. All of them belonged to HEV-B species. The study showed that echovirus 6 genotype accounted for 42% of the sequenced cases. The HEV infections in the present study tended to show slight seasonal pattern with more cases occurring during spring and summer, yet still significant numbers were also reported in fall and winter seasons.ConclusionHEV was isolated from a significant number of children with sepsis-like illness and/or aseptic meningitis. In addition, the molecular method utilized for direct diagnosis and genotyping of HEV from CSF revealed that more than one HEV type circulated in the West Bank, Palestine during the study period.

Genetic diversity of the enteroviruses detected from Cerebrospinal fluid (CSF) samples of patients with suspected aseptic meningitis in northern West Bank, Palestine in 2017

“…So far, only one report in Palestine confirmed the isolation of 7 different enteroviruses genotypes including echovirus 13, 14, 9, 30, 16, 6 and coxsackievirus B5 from sporadic cases of aseptic meningitis and/or sepsis like illness[10]. In the present study, all of the HEV positive cases occurred in children less than 7 years old and 50% of them < 1-year-old.…”

“…In the present study, all of the HEV positive cases occurred in children less than 7 years old and 50% of them < 1-year-old. Highest rate of enterovirus-positive cases was reported previously in children less than 1 year of age in Palestine, United States, and Korea [10, 38]. On the contrary, other studies, reported that HEV aseptic meningitis occurs most frequently in patients with age range of 3–12 years’ old [13-15, 39].…”

“…To identify the enterovirus associated aseptic meningitis cases, the HEV RNA was amplified using two primer sets targeting the 5’NCR of the HEV genome as described and modified previously[10, 21]. For HEV genotyping, an RT-PCR with two primer sets was used to amplify the 5’ half of the VP1region of the viral genome as described previously[10, 22]. PCR amplicons were purified using the NucleoSpin Gel and PCR Clean-up from Machery Nagel (Germany) before sending for commercial sequencing.…”

“…HEV-B causing aseptic meningitis include echoviruses, some enteroviruses coxsachievirus B[3-5]. Furthermore, a sever and potentially fatal conditions including encephalitis, acute flaccid paralysis, myocarditis and hand-foot-mouth disease had been reported [6-10].…”

“…In the last two decades, echovirus18 (E18) and coxsackievirus B5 (CVB5), serotypes of HEV-B, have been isolated from sporadic and outbreak cases of aseptic meningitis in China, Taiwan, Germany, Korea, Australia and Palestine. These isolates were found to be associated with various degrees of illness among newborns, infants and school-age children, ranging from asymptomatic to fatal aseptic meningitis [10-17].…”