Molecular Typing of Enteroviruses: Current Status and Future Requirements

Abstract

Human enteroviruses have traditionally been typed according to neutralization serotype. This procedure is limited by the difficulty in culturing some enteroviruses, the availability of antisera for serotyping, and the cost and technical complexity of serotyping procedures. Furthermore, the impact of information derived from enterovirus serotyping is generally perceived to be low. Enteroviruses are now increasingly being detected by PCR rather than by culture. Classical typing methods will therefore no longer be possible in most instances. An alternative means of enterovirus typing, employing PCR in conjunction with molecular genetic techniques such as nucleotide sequencing or nucleic acid hybridization, would complement molecular diagnosis, may overcome some of the problems associated with serotyping, and would provide additional information regarding the epidemiology and biological properties of enteroviruses. We argue the case for developing a molecular typing system, discuss the genetic basis of such a system, review the literature describing attempts to identify or classify enteroviruses by molecular methods, and suggest ways in which the goal of molecular typing may be realized.

Nucleotide Analysis and Phylogenetic Study of the Homology Boundaries of Coxsackie A and B Viruses

“…The disadvantages of this method are that it is time consuming, expensive and is often characterized by the inability to produce reliable results and to detect antigenic variants [1][2][3][4]. These drawbacks are associated with the existence of viral mixtures, antigenic drift and with the fact that the enteroviruses' serotypes detected by the intersecting pools of sera are limited (the actually used RIVM pools of sera detect only 42 of the 64 enteroviruses' serotypes) and have been raised against serotypes that have been isolated a long time ago (in the late 50s early 60s).…”

Molecular epidemiology of coxsackievirus type B1

“…Traditionally, CVB1, like other enteroviruses, has been detected by virus isolation in cell culture and subsequently serotyped by serum neutralizing assays with pools of serotype-specific antisera [10]. This method is time-consuming and labor intensive, and the availability of specific antisera becomes gradually restricted.…”

Isolation of polioviruses and other enteroviruses in south Greece between 1994 and 1998

“…Due to the frequently encountered low specificity and sensitivity of the currently available cell culture systems, the method of an enterovirus-specific reverse transcription-polymerase chain reaction was also used for the detection of enteroviruses in the clinical samples (5). After the observation of CPE, the infected cells were frozen at -80°C and thawed three times; 350 µl of the cell culture were taken and used for RNA extraction with the phenol-based TRIzol commercial kit by Gibco BRL (Life Technologies Ltd., Paisley, UK) according to the manufacturer's instructions.…”

“…Enteroviruses are the most commonly implicated agents of acute myocarditis and aseptic meningitis. Congenital infections also may occur, although their frequency is unknown and infection of neonate is frequently life-threatening (5). There is some evidence that enteroviruses are the possible etiological agents that cause or contribute to chronic diseases like dilated cardiomyopathy, one of the most common indications for heart transplantation, insulin-dependent diabetes mellitus, chronic fatigue syndrome and more, although such a role is still under debate (6)(7)(8).…”