Species-specific RT-PCR amplification of human enteroviruses: a tool for rapid species identification of uncharacterized enteroviruses.

نویسندگان

  • M Steven Oberste
  • Kaija Maher
  • Alford J Williams
  • Naomi Dybdahl-Sissoko
  • Betty A Brown
  • Michelle S Gookin
  • Silvia Peñaranda
  • Nada Mishrik
  • Moyez Uddin
  • Mark A Pallansch
چکیده

The 65 serotypes of human enteroviruses are classified into four species, Human enterovirus (HEV) A to D, based largely on phylogenetic relationships in multiple genome regions. The 3'-non-translated region of enteroviruses is highly conserved within a species but highly divergent between species. From this information, species-specific RT-PCR primers were developed that can be used to rapidly screen collections of enterovirus isolates to identify species of interest. The four primer pairs were 100 % specific when tested against enterovirus prototype strains and panels of isolates of known serotype (a total of 193 isolates). For evaluation in a typical application, the species-specific primers were used to screen 186 previously uncharacterized non-polio enterovirus isolates. The HEV-B primers amplified 68.3 % of isolates, while the HEV-A and HEV-C primers accounted for 9.7 and 11.3 % of isolates, respectively; no isolates were amplified with the HEV-D primers. Twelve isolates (6.5 %) were amplified by more than one primer set and eight isolates (4.3 %) were not amplified by any of the four primer pairs. Serotypes were identified by partial sequencing of the VP1 capsid gene, and in every case sequencing confirmed that the species-specific PCR result was correct; the isolates that were amplified by more than one species-specific primer pair were mixtures of two (11 isolates) or three (one isolate) species of viruses. The eight isolates that were not amplified by the species-specific primers comprised four new serotypes (EV76, EV89, EV90 and EV91) that appear to be unique members of HEV-A based on VP1, 3D and 3'-non-translated region sequences.

برای دانلود متن کامل این مقاله و بیش از 32 میلیون مقاله دیگر ابتدا ثبت نام کنید

ثبت نام

اگر عضو سایت هستید لطفا وارد حساب کاربری خود شوید

منابع مشابه

Direct Identification of Enteroviruses in Cerebrospinal Fluid of Patients with Suspected Meningitis by Nested PCR Amplification

Enteroviruses, the most common human viral pathogens worldwide, have been associated with serous meningitis, encephalitis, syndrome of acute flaccid paralysis, myocarditis and the onset of diabetes type 1. In the future, the rapid identification of the etiological agent would allow to adjust the therapy promptly and thereby improve the course of the disease and prognosis. We developed RT-nested...

متن کامل

Evaluation of Nucleic Acid Sequence Based Amplification (NASBA) and Reverse Transcription Polymerase Chain Reaction for Detection of Coxsackievirus B3 in Cell Culture and Animal Tissue Samples

Enteroviruses are the causative agents of a number of diseases in humans. Group B coxsackieviruses are believed to be the most common viral agents responsible for human heart disease. Genomic data of enteroviruses has allowed developing new molecular approaches such as Nucleic Acid Sequence Based Amplification (NASBA) for detection of such viruses. In this study, coxsackievirus B3 (CVB3) was de...

متن کامل

تشخیص همزمان ویروس‌های انترو و اوریون در نمونه‌های مایع مغزی نخاعی مشکوک به مننژیت توسط تست RT-PCR

  Introduction: Detection of enteroviruses and mumps virus nucleic acid in cerebrospinal spinal fluid (CSF) specimens has been demonstrated to improve the management of patients with aseptic meningitis. Then rapid and sensitive diagnostic techniques are desirable.   Materials & Methods: Multiplex RT-PCR has the potential to detection both of viruses. This assay is so rapid and sensitive. At fir...

متن کامل

Characterization of the genome of human enteroviruses: design of generic primers for amplification and sequencing of different regions of the viral genome.

Human enteroviruses are among the most common viruses infecting humans and can cause diverse clinical syndromes ranging from minor febrile illness to severe and potentially fatal diseases. Biodiversity and evolution of human enterovirus genomes are shaped by frequent recombination events. Therefore, identification and characterization of circulating strains of enteroviruses require partial dete...

متن کامل

The Association of Non-polio Enteroviruses with Aseptic Meningitis in Children in Iran

  Introduction : Viral meningitis is an inflammation of the leptomeninges as a manifestation of central nervous system (CNS) infection, and more than 85% of viral meningitis cases are caused by non-polio enteroviruses. Molecular methods such as polymerase chain reaction (PCR) are increasingly used to detect and type non-polio enteroviruses. Here, we describe the impact of these viruses on asept...

متن کامل

ذخیره در منابع من


  با ذخیره ی این منبع در منابع من، دسترسی به آن را برای استفاده های بعدی آسان تر کنید

برای دانلود متن کامل این مقاله و بیش از 32 میلیون مقاله دیگر ابتدا ثبت نام کنید

ثبت نام

اگر عضو سایت هستید لطفا وارد حساب کاربری خود شوید

عنوان ژورنال:
  • The Journal of general virology

دوره 87 Pt 1  شماره 

صفحات  -

تاریخ انتشار 2006