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.

FIG. 1

Performance of participating laboratories in detecting enterovirus RNA dilutions included in panel A samples. The datum sets obtained by the Roche Amplicor test are shown as white columns, while the datum sets obtained by in-house single PCR or nested PCR assays are shown as diagonally striped and cross-hatched columns, respectively. A positive result is indicated by a raised column.

FIG. 2

Performance of participating laboratories in detecting enterovirus in different specimen types included in panel B samples. See legend to Fig. 1 for interpretation of the columns.

FIG. 3

Performance of participating laboratories in detecting enterovirus serotypes included in panel C samples. See legend to Fig. 1 for interpretation of the columns. Samples identified in black were not tested.