Enterovirus Detection in Patients Suspected Aseptic Meningitis by RT-PCR in Kermanshah, Iran

Document Type: Original Article

Authors

1 Associate Professor of Medical Microbiology, Nosocomial Infection Research Centre, School of Medicine, Kermanshah University of Medical Sciences, Kermanshah, Iran

2 MSc Student of Medical Microbiology, Student Research Committee, Faculty of Medicine, Kermanshah University of Medical Sciences, Kermanshah, Iran

3 Ph.D. Candidate, Department of Microbiology, Faculty of Medicine, Kermanshah University of Medical Sciences, Kermanshah, Iran

4 Assistant Professor, Department of Virology Faculty of public health, Tehran University of medical sciences, Tehran, Iran

5 Assistant Professor, Department of Microbiology, Faculty of Medicine, Kermanshah University of Medical Sciences, Kermanshah, Iran

Abstract

Background: Aseptic meningitis is frequently caused by viral agents, particularly human enterovirus. Several methods of Reverse Transcription PCR(RT-PCR) have recently been introduced and modified for better diagnosis of enteroviral infection in meningitis. This study aimed to determine enteroviruses in patients with suspicion of aseptic meningitis using RT-PCR in the West of Iran.
Methods: In this study, 120 CSF samples were collected from patients hospitalized with the suspicion of aseptic meningitis in Imam Reza Hospital of Kermanshah, Iran.RT-PCR was used to diagnose enteroviruses. The cDNA recovered from RT-PCR was purified using a DNA purification kit and sequenced to confirm viral genome. Sequence data were analyzed for homology using the Gen Bank database.
Results: The samples were collected from 63 (52.5%) men and 57 (47.5%) women with an average age of 31.5 ± 29.4 years. Of the samples tested, 4 cases (3.33%) yielded positive results for enterovirus. The results of sequence data analysis confirmed all positive cases as Enterovirus type 71. The biochemical (protein and glucose) and cytological analyses of positive CSF samples showed no significant changes.
Conclusion: According to the results, Enterovirus type 71is one of the common causes of enteroviral meningitis in patients in Kermanshah. The RT-PCR technique can rapidly diagnose Enterovirus type 71 and help better treatment of the patients and prevent the unnecessary use of antibiotics. So, molecular methods can reduce the cost of patients’ treatments and prevent drug resistance among bacteria. It can also provide a better picture of enteroviral infection in our region.

Keywords


 

  1. Dalwai A, Ahmad S, Al-Nakib W. Echo viruses are a major cause of aseptic meningitis in infants and young children in Kuwait. Virology journal. 2010(7):236.
  2. Rasti M, Samarbaf-Zadeh AR, Makvandi M, Shamsi-Zadeh A. Relative Frequency of Enteroviruses in Children With Aseptic Meningitis Referred to Aboozar Hospital in Ahvaz Jundishapur. Jundishapur J Microbiol. 2012; 5(3):456-9.
  3. Brunel D, Leveque N, Jacques J, Renois F, Motte J, Andreoletti L. Clinical and virological features of an aseptic meningitis outbreak in North-Eastern France, 2005. Journal of clinical virology : the official publication of the Pan American Society for Clinical Virology. 2008; 42(2):225-8.
  4. Sadeuh-Mba SA, Bessaud M, Massenet D, Joffret ML, Endegue MC, Njouom R, et al. High frequency and diversity of species C enteroviruses in Cameroon and neighboring countries. Journal of clinical microbiology. 2013; 51(3):759-70.
  5. Simmonds P, Welch J. Frequency and Dynamics of Recombination within Different Species of Human Enteroviruses. J Virol. 2006; 80(1):483-93.
  6. Dierssen U, Rehren F, Henke-Gendo C, Harste G, Heim A. Rapid routine detection of enterovirus RNA in cerebrospinal fluid by a one-step real-time RT-PCR assay. J Clin Virol. 2008; 42:58-64.
  7. Hyeon JY, Hwang S, Kim H, Song J, Ahn J, Kang B, et al. Accuracy of diagnostic methods and surveillance sensitivity for human enterovirus, South Korea, 1999-2011. Emerging infectious diseases. 2013; 19(8):1268-75.
  8. Tong CYW, Bible JM. Global epidemiology of enterovirus 71. Future Virology 2009; 4(5):501-10.
  9. Perez-Velez CM, Anderson MS, Robinson CC, McFarland EJ, Nix WA, Pallansch MA, et al. Outbreak of neurologic enterovirus type 71 disease: a diagnostic challenge. Clinical infectious diseases : an official publication of the Infectious Diseases Society of America. 2007; 45(8):950-7.
  10. Kargar M, Sadeghipour S, Nategh R. Environmental surveillance of Non-Polio Enteroviruses in Iran. Virology journal. 2009; 6:149.
  11. Archimbaud C, Chambon M, Bailly JL, Petit I, Henquell C, Mirand A, et al. Impact of rapid enterovirus molecular diagnosis on the management of infants, children, and adults with aseptic meningitis. Journal of medical virology. 2009; 81(1):42-8.
  12. Allan Nix W, Steven Oberste M, Pallansch MA. Sensitive, Seminested PCR Amplification of VP1 Sequences for Direct Identification of All Enterovirus Serotypes from Original Clinical Specimens. Journal of clinical microbiology. 2006; 44(8):2698-704.
  13. Martinez Planas A, Munoz Almagro C, Luaces Cubells C, Noguera Julian A, Selva L, Fernandez JP, et al. Low prevalence of invasive bacterial infection in febrile infants under 3 months of age with enterovirus infection. Clinical microbiology and infection : the official publication of the European Society of Clinical Microbiology and Infectious Diseases. 2012; 18(9):856-61.
  14. Van Loon AM, Cleator GC, Ras A. External quality assessment of enterovirus detection and typing. European Union Concerted Action on Virus Meningitis and Encephalitis. Bulletin of the World Health Organization. 1999; 77(3):217-23.
  15. Giri A, Arjyal A, Koirala S, Karkey A, Dongol S, Thapa SD, et al. Aetiologies of central nervous system infections in adults in Kathmandu, Nepal: a prospective hospital-based study. Scientific reports. 2013; 3:2382.
  16. Meqdam MM, Khalousi MM, Al-Shurman A. Enteroviral meningitis in Northern Jordan: prevalence and association with clinical findings. Journal of medical virology. 2002; 66(2):224-8.
  17. Racaniello VR. Picornaviridae: the viruses and their replication 2001.
  18. McMinn PC. An overview of the evolution of enterovirus 71 and its clinical and public health significance. FEMS microbiology reviews. 2002; 26(1):91-107.
  19. Perera D, Podin Y, Akin W, Tan CS, Cardosa MJ. Incorrect identification of recent Asian strains of Coxsackievirus A16 as human enterovirus 71: improved primers for the specific detection of human enterovirus 71 by RT PCR. BMC infectious diseases. 2004; 4:11.
  20. Ho Dang Trung N, Le Thi Phuong T, Wolbers M, Nguyen Van Minh H, Nguyen Thanh V, Pham Van M, et al. A etiologies of central nervous system infection in Viet Nam: a prospect ive provincial hospital-based descriptive surveilla nce study. PLoS One 7. 2012; 7(5):e37825.
  21. Kupila L, Vuorinen T, Vainionpaa R, Hukkanen V, Marttila RJ, Kotilainen P. Etiology of aseptic meningitis and encephalitis in an adult population. Neurology. 2006; 66(1):75-80.
  22. Verboon-Maciolek MA, Nijhuis M, van Loon AM, van Maarssenveen N, van Wieringen H, Pekelharing-Berghuis MA, et al. Diagnosis of enterovirus infection in the first 2 months of life by real-time polymerase chain reaction. Clinical infectious diseases : an official publication of the Infectious Diseases Society of America. 2003; 37(1):1-6.
  23. Mamishi S, Rahimi P, Sohrabi A, Sabuni F, Edalat R, Mostafavi E, et al. Direct Serotyping of Enteroviruses in Cerebrospinal Fluid of Children With Aseptic Meningitis. Jundishapur J Microbiol. 2013 6(9):e7852.
  24. Roohandeh A, Rahimi P, Sohrabi A, Mobasheri M, Azadmanesh K, Shahosseini Z, et al. Frequency of human enterovirus 71 in children under 8 years old with aseptic menengitis in Tehran. Clinical laboratory. 2013;59(7-8):915-20.
  25. Hosseininasab A, Alborzi A, Ziyaeyan M, Jamalidoust M, Moeini M, Pouladfar G, et al. Viral etiology of aseptic meningitis among children in southern Iran. Journal of medical virology. 2011;83(5):884-8.
  26. Seo JH, Yeom JS, Youn HS, Han TH, Chung JY. Prevalence of human parechovirus and enterovirus in cerebrospinal fluid samples in children in Jinju, Korea. Korean journal of pediatrics. 2015; 58(3):102-7.
  27. Vuorinen T, Vainionpaa R, Hyypia T. Five years' experience of reverse-transcriptase polymerase chain reaction in daily diagnosis of enterovirus and rhinovirus infections. Clinical infectious diseases : an official publication of the Infectious Diseases Society of America. 2003; 37(3):452-5.
  28. Antona D, Leveque N, Chomel JJ, Dubrou S, Levy-Bruhl D, Lina B. Surveillance of enteroviruses in France, 2000-2004. European journal of clinical microbiology & infectious diseases : official publication of the European Society of Clinical Microbiology. 2007; 26(6):403-12.
  29. Rice P. Viral meningitis and encephalitis. Elsevier Ltd. 2013; 41(12):678-82.
  30. Kim HJ, Kang B, Hwang S, Hong J, Kim K, Cheon DS. Epidemics of viral meningitis caused by echovirus 6 and 30 in Korea in 2008. Virology journal. 2012; 9:38.
  31. Menasalvas-Ruiz AI, Salvador-Garcia C, Moreno-Docon A, Alfayate-Miguelez S, Perez Canovas C, Sanchez-Solis M. Enterovirus reverse transcriptase polymerase chain reaction assay in cerebrospinal fluid: An essential tool in meningitis management in childhood. Enferm Infecc Microbiol Clin. 2013; 31(2):71-5.