Multicenter Quality Assessment of PCR Methods for Detection of Enteroviruses

Abstract

We conducted a multicenter evaluation of commercial and in-house PCR methods for the detection of enteroviruses. Three coded panels of test and control RNA samples, artificial clinical specimens, and representative enterovirus serotypes were used to assess amplification methods, RNA extraction methods, and reactivities with different enterovirus serotypes. Despite several differences between PCR methods, there was good agreement, although some variation in sensitivity was observed. Most PCR methods were able to detect enterovirus RNA derived from 0.01 50% tissue culture infective dose (TCID50) and were able to detect at least 1 TCID50 of enterovirus in cerebrospinal fluid, stool, or throat swab specimens. Most were also able to detect a wide range of enterovirus serotypes, although serotypic identification was not possible. Some laboratories experienced false-positive results due to PCR contamination, which appeared to result mainly from cross-contamination of specimens during RNA extraction. Provided that this problem is overcome, these PCR methods will prove to be a sensitive and rapid alternative to cell culture for the diagnosis of enterovirus infection.

Occurrence of bacterial indicators and bacteriophages infecting enteric bacteria in groundwater in different geographical areas

“…Moreover, several multicentre studies for the detection of viruses by genomic methods revealed problems of standardization (Lina et al. 1996; Muir et al. 1999), which increase still further with water samples because of the need for a virus concentration step.…”

Enterovirus meningitis in adults in 1999–2000 and evaluation of clinical management

“…Enteroviruses,particularlycoxsackievirusesandechoviruses, are the most common cause of aseptic meningitis in the paediatric population [Romero, 1999]. Detection of the enterovirus genome in cerebrospinal uid (CSF) has proven conclusively to be useful and more rapid and sensitive than viral culture [Andre Âoletti et al, 1998;Muir et al, 1999;Romero, 1999]. Moreover, early detection of a positive enterovirus genome using reverse transcription-polymerase chain reaction (RT-PCR) can save considerable health resources by reducing the duration of antibiotic or antiviral therapy, avoiding unnecessary diagnosis interventions, and allowing rapid discharge of patients [Romero, 1999;Ramers et al, 2000;Spicher et al, 2000], as seen in paediatric patients presenting with enterovirus meningitis.…”

Molecular analysis of cerebrospinal fluid in viral diseases of the central nervous system

“…To help obviate these problems, quality control (QC) programmes are being carried out for viruses and other infectious agents. QC assessments for viruses responsible of CNS infections, including enteroviruses, HSV-1, HSV-2 and JCV, have been performed as a part of European Union sponsored QC in virology programmes (van Vliet et al, 1998;Muir et al, 1999;van Loon et al, 1999;Weber et al, 1997;Schloss et al, 2001). These are based on the use of panels consisting of coded samples containing known amounts of NA molecules and control samples, which are distributed to participant laboratories and therein tested blindly.…”

“…Analysis of reported results has commonly revealed substantial different reports between laboratories, especially with samples containing low amounts of NA. Furthermore, a significant rate of false positive results has been observed (Muir et al, 1999;Schloss et al, 2001). On the other hand, little or no relationship is generally found between performance and the use of in-house rather than commercial techniques.…”